98 results on '"Kalra, S."'
Search Results
2. Impact of aspirin use on rates of metastasis in patients with esophageal cancer: insights from the National Inpatient Sample.
- Author
-
Patel J, Khanna T, Sohal A, Dhaliwal A, Chaudhry H, Kalra S, Singh I, Dukovic D, and Bains K
- Subjects
- Humans, Male, Female, Aged, Middle Aged, United States epidemiology, Comorbidity, Neoplasm Metastasis, Adult, Inpatients statistics & numerical data, Retrospective Studies, Gastrointestinal Neoplasms pathology, Logistic Models, Aspirin therapeutic use, Esophageal Neoplasms pathology
- Abstract
Despite advancing treatment methods, esophageal cancer (EC) maintains a high mortality rate and poor prognosis. Through various mechanisms, aspirin has been suggested to have a chemopreventive effect on EC. However, the long-term impact, particularly regarding the rate of metastasis, needs to be further elucidated. NIS 2016-2020 was used to identify adult patients (age > 18 years) with EC using ICD-10 codes. Patients with missing demographics and mortality were excluded. Patients were stratified into two groups based on aspirin use. Data were collected on patient demographics, Elixhauser Comorbidity Index (ECI), and comorbidities (hypertension, chronic pulmonary disease, coronary artery disease (CAD), chronic kidney disease (CKD), congestive heart failure (CHF), coagulopathy, alcohol use, smoking, and obesity). The outcomes studied were rates of total metastasis, gastrointestinal (GI) metastasis, non-GI metastasis, and lymphoid metastasis. Multivariate logistic regression analysis was performed to evaluate the impact of aspirin use on various metastases after adjusting for patient demographics, comorbidities, and ECI. Out of 190,655 patients, 20,650 (10.8%) patients were aspirin users. Majority of the patients in the aspirin group were aged > 65 years (74.7%), males (82.1%), White race (84%), and had medicare insurance (71%). There was a higher incidence of diabetes, hypertension, chronic pulmonary disease, CAD, CKD, CHF, and smoking in aspirin users than non-aspirin users. Patients with aspirin users had a lower incidence of metastasis (28.9% vs. 38.7%, P < 0.001), GI metastasis (14.2% vs. 20.6%, P < 0.001), non-GI metastasis (15.1% vs. 22%, P < 0.001), and lymphoid metastasis (8.9% vs. 11.3%, P < 0.001) than non-aspirin users. After adjusting for confounding factors, patients with aspirin use had lower odds of having metastasis (aOR-0.73, 95% CI-0.70-0.77, P < 0.001). Our study noted that aspirin use is associated with a reduction in the rate of metastasis in patients with EC. These studies support the use of aspirin in patients with EC and suggest the need for further studies to understand the mechanism by which aspirin use reduces metastasis in patients with EC., (© The Author(s) 2024. Published by Oxford University Press on behalf of International Society for Diseases of the Esophagus. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2024
- Full Text
- View/download PDF
3. Global Barriers to Accessing Off-Patent Endocrine Therapies: A Renaissance of the Orphan Disease?
- Author
-
de Silva NL, Dissanayake H, Kalra S, Meeran K, Somasundaram NP, and Jayasena CN
- Subjects
- Humans, Costs and Cost Analysis, Drugs, Generic therapeutic use, Drug Costs, Rare Diseases drug therapy
- Abstract
Context: Clinical endocrinology encompasses many diseases requiring long-term drug therapy. Prohibitive pricing of some endocrine drugs classified as essential by the World Health Organization has created suboptimal care of patients with endocrine disorders., Evidence Acquisition: This review is based on evidence obtained from several databases and search engines including PubMed, Google, and Google Scholar; reference searches; manual searching for web pages of international regulatory bodies; and the authors' experience from different healthcare settings., Evidence Synthesis: After the expiry of a patent, generic versions with the opportunity for increased availability and a price reduction are expected. There are access barriers worldwide for many off-patent endocrine drugs. The high price is the main issue for several medicines including insulin, hydrocortisone, testosterone, and gonadotropins. This is caused by several factors including the market monopoly due to the lack of registered generics or suppliers limiting the benefit of competition and a complex supply chain. Additionally, the lack of some medicines has been concerning due to market factors such as the relatively small number of patients, making it less attractive for the manufacturers. Commissioning of nonprofit manufacturers and state manufacturing as well as strict price control measures could alleviate this situation., Conclusion: Lack of availability and disproportionate price inflation affecting essential off-patent endocrine therapies is common due to several interrelated factors. Global collaboration among healthcare organizations with the support of policymaking bodies might be needed to mitigate this., (© The Author(s) 2023. Published by Oxford University Press on behalf of the Endocrine Society.)
- Published
- 2024
- Full Text
- View/download PDF
4. Geriatric care physicians' perspectives on providing virtual care: a reflexive thematic synthesis of their online survey responses from Ontario, Canada.
- Author
-
Chuen VL, Dholakia S, Kalra S, Watt J, Wong C, and Ho JM
- Subjects
- Humans, Aged, Ontario, Pandemics, Surveys and Questionnaires, Physicians psychology, Telemedicine
- Abstract
Background: During the COVID-19 pandemic, telemedicine was widely implemented to minimise viral spread. However, its use in the older adult patient population was not well understood., Objective: To understand the perspectives of geriatric care providers on using telemedicine with older adults through telephone, videoconferencing and eConsults., Design: Qualitative online survey study., Setting and Participants: We recruited geriatric care physicians, defined as those certified in Geriatric Medicine, Care of the Elderly (family physicians with enhanced skills training) or who were the most responsible physician in a long-term care home, in Ontario, Canada between 22 December 2020 and 30 April 2021., Methods: We collected participants' perspectives on using telemedicine with older adults in their practice using an online survey. Two researchers jointly analysed free-text responses using the 6-phase reflexive thematic analysis., Results: We recruited 29 participants. Participants identified difficulty using technology, patient sensory impairment, lack of hospital support and pre-existing high patient volumes as barriers against using telemedicine, whereas the presence of a caregiver and administrative support were facilitators. Perceived benefits of telemedicine included improved time efficiency, reduced travel, and provision of visual information through videoconferencing. Ultimately, participants felt telemedicine served various purposes in geriatric care, including improving accessibility of care, providing follow-up and obtaining collateral history. Main limitations are the absence of, or incomplete physical exams and cognitive testing., Conclusions: Geriatric care physicians identify a role for virtual care in their practice but acknowledge its limitations. Further work is required to ensure equitable access to virtual care for older adults., (© The Author(s) 2024. Published by Oxford University Press on behalf of the British Geriatrics Society. All rights reserved. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2024
- Full Text
- View/download PDF
5. Resurfacing in Facial Burn Sequelae Using Parascapular Free Flap: A Long-Term Experience.
- Author
-
Kalra GS, Kalra S, and Gupta S
- Subjects
- Cicatrix etiology, Cicatrix surgery, Humans, Retrospective Studies, Burns complications, Burns surgery, Facial Injuries surgery, Free Tissue Flaps, Neck Injuries surgery, Plastic Surgery Procedures methods
- Abstract
It is difficult to treat large postburn sequelae (scars and defects) over the face. Available methods include skin grafts, local flaps, tissue expansion, and free flaps. These surgical options should be chosen wisely, depending on individual patient requirements and the area involved. In patients with large postburn scars and defects in which the surrounding tissue is also involved, use of free tissue transfer is extremely useful. A retrospective analysis was done between 2011 and 2019 of 52 cases with extensive facial burn deformities in whom secondary reconstruction was done with free parascapular flap cover in our department. Outcome was assessed by a direct questionnaire. There was no complete flap loss in the series. Two cases were re-explored for venous insufficiency and suffered partial marginal necrosis. Twenty patients had to undergo further debulking procedure. Forty-seven patients were satisfied by the final outcome. Postburn facial deformities are difficult to treat; in many cases, there are no local options, and tissue from different regions is to be used for reconstruction. Free parascapular flaps can be used as an effective method in such cases with a high level of patient satisfaction., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Burn Association. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2022
- Full Text
- View/download PDF
6. EcTracker: Tracking and elucidating ectopic expression leveraging large-scale scRNA-seq studies.
- Author
-
Gautam V, Mittal A, Kalra S, Mohanty SK, Gupta K, Rani K, Naidu S, Mishra T, Sengupta D, and Ahuja G
- Subjects
- Databases, Genetic, Gene Expression Profiling, Gene Regulatory Networks, Humans, Organ Specificity, Transcription Factors metabolism, User-Computer Interface, Web Browser, Computational Biology methods, Ectopic Gene Expression, RNA-Seq, Single-Cell Analysis methods, Software
- Abstract
Dramatic genomic alterations, either inducible or in a pathological state, dismantle the core regulatory networks, leading to the activation of normally silent genes. Despite possessing immense therapeutic potential, accurate detection of these transcripts is an ever-challenging task, as it requires prior knowledge of the physiological gene expression levels. Here, we introduce EcTracker, an R-/Shiny-based single-cell data analysis web server that bestows a plethora of functionalities that collectively enable the quantitative and qualitative assessments of bona fide cell types or tissue-specific transcripts and, conversely, the ectopically expressed genes in the single-cell ribonucleic acid sequencing datasets. Moreover, it also allows regulon analysis to identify the key transcriptional factors regulating the user-selected gene signatures. To demonstrate the EcTracker functionality, we reanalyzed the CRISPR interference (CRISPRi) dataset of the human embryonic stem cells differentiated into endoderm lineage and identified the prominent enrichment of a specific gene signature in the SMAD2 knockout cells whose identity was ambiguous in the original study. The key distinguishing features of EcTracker lie within its processing speed, availability of multiple add-on modules, interactive graphical user interface and comprehensiveness. In summary, EcTracker provides an easy-to-perform, integrative and end-to-end single-cell data analysis platform that allows decoding of cellular identities, identification of ectopically expressed genes and their regulatory networks, and therefore, collectively imparts a novel dimension for analyzing single-cell datasets., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2021
- Full Text
- View/download PDF
7. Interventional cardiology's cross-departmental effort with production of reusable personal protective equipment during the early phase of COVID-19 pandemic.
- Author
-
Memon S, Friend E, George JC, and Kalra S
- Subjects
- Humans, Pandemics, Personal Protective Equipment, SARS-CoV-2, COVID-19, Cardiology
- Published
- 2021
- Full Text
- View/download PDF
8. Machine-OlF-Action: a unified framework for developing and interpreting machine-learning models for chemosensory research.
- Author
-
Gupta A, Choudhary M, Mohanty SK, Mittal A, Gupta K, Arya A, Kumar S, Katyayan N, Dixit NK, Kalra S, Goel M, Sahni M, Singhal V, Mishra T, Sengupta D, and Ahuja G
- Abstract
Summary: Machine Learning-based techniques are emerging as state-of-the-art methods in chemoinformatics to selectively, effectively and speedily identify biologically relevant molecules from large databases. So far, a multitude of such techniques have been proposed, but unfortunately due to their sparse availability, and the dependency on high-end computational literacy, their wider adaptation faces challenges, at least in the context of G-Protein Coupled Receptors (GPCRs)-associated chemosensory research. Here, we report Machine-OlF-Action (MOA), a user-friendly, open-source computational framework, that utilizes user-supplied SMILES (simplified molecular input line entry system) of the chemicals, along with their activation status, to synthesize classification models. MOA integrates a number of popular chemical databases collectively harboring approximately 103 million chemical moieties. MOA also facilitates customized screening of user-supplied chemical datasets. A key feature of MOA is its ability to embed molecules based on the similarity of their local neighborhood, by utilizing a state-of-the-art model interpretability framework LIME. We demonstrate the utility of MOA in identifying previously unreported agonists for human and mouse olfactory receptors OR1A1 and MOR174-9 by leveraging the chemical features of their known agonists and non-agonists. In summary, here we develop an ML-powered software playground for performing supervisory learning tasks involving chemical compounds., Availability and Implementation: MOA is available for Windows, Mac and Linux operating systems. It's accessible at (https://ahuja-lab.in/). Source code, user manual, step-by-step guide and support is available at GitHub (https://github.com/the-ahuja-lab/Machine-Olf-Action). For results, reproducibility and hyperparameters, refer to Supplementary Notes., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2021
- Full Text
- View/download PDF
9. The Cellular basis of loss of smell in 2019-nCoV-infected individuals.
- Author
-
Gupta K, Mohanty SK, Mittal A, Kalra S, Kumar S, Mishra T, Ahuja J, Sengupta D, and Ahuja G
- Subjects
- Angiotensin-Converting Enzyme 2 metabolism, COVID-19 pathology, COVID-19 virology, Humans, SARS-CoV-2 isolation & purification, Viral Proteins metabolism, Virus Internalization, Anosmia pathology, COVID-19 complications
- Abstract
A prominent clinical symptom of 2019-novel coronavirus (nCoV) infection is hyposmia/anosmia (decrease or loss of sense of smell), along with general symptoms such as fatigue, shortness of breath, fever and cough. The identity of the cell lineages that underpin the infection-associated loss of olfaction could be critical for the clinical management of 2019-nCoV-infected individuals. Recent research has confirmed the role of angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2) as key host-specific cellular moieties responsible for the cellular entry of the virus. Accordingly, the ongoing medical examinations and the autopsy reports of the deceased individuals indicate that organs/tissues with high expression levels of ACE2, TMPRSS2 and other putative viral entry-associated genes are most vulnerable to the infection. We studied if anosmia in 2019-nCoV-infected individuals can be explained by the expression patterns associated with these host-specific moieties across the known olfactory epithelial cell types, identified from a recently published single-cell expression study. Our findings underscore selective expression of these viral entry-associated genes in a subset of sustentacular cells (SUSs), Bowman's gland cells (BGCs) and stem cells of the olfactory epithelium. Co-expression analysis of ACE2 and TMPRSS2 and protein-protein interaction among the host and viral proteins elected regulatory cytoskeleton protein-enriched SUSs as the most vulnerable cell type of the olfactory epithelium. Furthermore, expression, structural and docking analyses of ACE2 revealed the potential risk of olfactory dysfunction in four additional mammalian species, revealing an evolutionarily conserved infection susceptibility. In summary, our findings provide a plausible cellular basis for the loss of smell in 2019-nCoV-infected patients., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)
- Published
- 2021
- Full Text
- View/download PDF
10. A practical risk score for early prediction of neurological outcome after out-of-hospital cardiac arrest: MIRACLE2.
- Author
-
Pareek N, Kordis P, Beckley-Hoelscher N, Pimenta D, Kocjancic ST, Jazbec A, Nevett J, Fothergill R, Kalra S, Lockie T, Shah AM, Byrne J, Noc M, and MacCarthy P
- Subjects
- Aged, Aged, 80 and over, Humans, Middle Aged, Prognosis, Registries, Risk Factors, Cardiopulmonary Resuscitation, Out-of-Hospital Cardiac Arrest
- Abstract
Aims: The purpose of this study was to develop a practical risk score to predict poor neurological outcome after out-of-hospital cardiac arrest (OOHCA) for use on arrival to a Heart Attack Centre., Methods and Results: From May 2012 to December 2017, 1055 patients had OOHCA in our region, of whom 373 patients were included in the King's Out of Hospital Cardiac Arrest Registry (KOCAR). We performed prediction modelling with multivariable logistic regression to identify predictors of the primary outcome to derive a risk score. This was externally validated in two independent cohorts comprising 473 patients. The primary endpoint was poor neurological outcome at 6-month follow-up (Cerebral Performance Category 3-5). Seven independent predictors of outcome were identified: missed (unwitnessed) arrest, initial non-shockable rhythm, non-reactivity of pupils, age (60-80 years-1 point; >80 years-3 points), changing intra-arrest rhythms, low pH <7.20, and epinephrine administration (2 points). The MIRACLE2 score had an area under the curve (AUC) of 0.90 in the development and 0.84/0.91 in the validation cohorts. Three risk groups were defined-low risk (MIRACLE2 ≤2-5.6% risk of poor outcome); intermediate risk (MIRACLE2 of 3-4-55.4% of poor outcome); and high risk (MIRACLE2 ≥5-92.3% risk of poor outcome). The MIRACLE2 score had superior discrimination than the OHCA [median AUC 0.83 (0.818-0.840); P < 0.001] and Cardiac Arrest Hospital Prognosis models [median AUC 0.87 (0.860-0.870; P = 0.001] and equivalent performance with the Target Temperature Management score [median AUC 0.88 (0.876-0.887); P = 0.092]., Conclusions: The MIRACLE2 is a practical risk score for early accurate prediction of poor neurological outcome after OOHCA, which has been developed for simplicity of use on admission., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2020. For permissions, please email: journals.permissions@oup.com.)
- Published
- 2020
- Full Text
- View/download PDF
11. Outcome of inter-hospital transfer versus direct admission for primary percutaneous coronary intervention: An observational study of 25,315 patients with ST-elevation myocardial infarction from the London Heart Attack Group.
- Author
-
Rathod KS, Jain AK, Firoozi S, Lim P, Boyle R, Nevett J, Dalby MC, Kalra S, Malik IS, Sirker A, Mathur A, Redwood S, MacCarthy PA, Wragg A, and Jones DA
- Subjects
- Female, Follow-Up Studies, Humans, Incidence, London epidemiology, Male, Middle Aged, Retrospective Studies, ST Elevation Myocardial Infarction epidemiology, Survival Rate trends, Hospitalization statistics & numerical data, Patient Admission statistics & numerical data, Patient Transfer statistics & numerical data, Percutaneous Coronary Intervention methods, Registries, ST Elevation Myocardial Infarction surgery
- Abstract
Background and Aims: In patients with ST-segment elevation myocardial infarction (STEMI), mortality is directly related to time to reperfusion with guidelines recommending patients be delivered directly to centres for primary percutaneous coronary intervention (PCI). The aim of this study was to describe the impact of inter-hospital transfer on reperfusion time and to assess whether or not treatment delays influenced clinical outcomes in comparison with direct admission to a primary PCI centre in a large regional network., Method and Results: We undertook an observational cohort study of patients with STEMI treated with primary PCI between 2005 and 2015 in London, UK. Patient details were recorded at the time of the procedure in databases using the British Cardiovascular Intervention Society PCI dataset. The primary end-point was all-cause mortality at a median of 4.1 years (interquartile range: 2.2-5.8 years). Secondary outcomes were in-hospital major adverse cardiac events. Of 25,315 patients, 17,560 (69.4%) were admitted directly to a primary PCI centre and 7755 (31.6%) were transferred from a non-primary PCI centre. Patients in the direct admission group were older and more likely to have left ventricular impairment compared with the inter-hospital transfer group. Median time from call for help to reperfusion in transferred patients was 52 minutes longer compared with patients admitted directly ( p <0.001). However, call to first hospital admission was similar. Kaplan-Meier analysis demonstrated significantly lower mortality rates in patients who were transferred directed to a primary PCI centre compared with patients who were transferred from a non-PCI centre (17.4% direct vs . 18.7% transfer, p =0.017). Furthermore, after propensity matching, direct admission for primary PCI was still a predictor of all-cause mortality (hazard ratio: 0.89, 95% confidence interval: 0.64-0.95)., Conclusions: In this large registry of over 25,000 STEMI patients treated by primary PCI survival was better in patients admitted directly to a cardiac centre versus patients transferred for primary PCI, most likely due to longer call to balloon times in patient transferred from other hospitals.
- Published
- 2020
- Full Text
- View/download PDF
12. Cerebral atrophy in amyotrophic lateral sclerosis parallels the pathological distribution of TDP43.
- Author
-
Dadar M, Manera AL, Zinman L, Korngut L, Genge A, Graham SJ, Frayne R, Collins DL, and Kalra S
- Abstract
Amyotrophic lateral sclerosis is a neurodegenerative disease characterized by a preferential involvement of both upper and lower motor neurons. Evidence from neuroimaging and post-mortem studies confirms additional involvement of brain regions extending beyond the motor cortex. The aim of this study was to assess the extent of cerebral disease in amyotrophic lateral sclerosis cross-sectionally and longitudinally and to compare the findings with a recently proposed disease-staging model of amyotrophic lateral sclerosis pathology. Deformation-based morphometry was used to identify the patterns of brain atrophy associated with amyotrophic lateral sclerosis and to assess their relationship with clinical symptoms. Longitudinal T
1 -weighted MRI data and clinical measures were acquired at baseline, 4 months and 8 months, from 66 patients and 43 age-matched controls who participated in the Canadian Amyotrophic Lateral Sclerosis Neuroimaging Consortium study. Whole brain voxel-wise mixed-effects modelling analysis showed extensive atrophy patterns differentiating patients from the normal controls. Cerebral atrophy was present in the motor cortex and corticospinal tract, involving both grey matter and white matter, and to a lesser extent in non-motor regions. More specifically, the results showed significant bilateral atrophy in the motor cortex and corticospinal tract (including the internal capsule and brainstem) and ventricular enlargement, along with significant longitudinal atrophy in precentral gyrus, frontal and parietal white matter, accompanied by ventricular and sulcal enlargement. Atrophy in the precentral gyrus was significantly associated with greater disability as quantified with the Amyotrophic Lateral Sclerosis Functional Rating Scale-Revised ( P < 0.0001). The pattern of atrophy observed using deformation-based morphometry was consistent with the Brettschneider's four-stage pathological model of the disease. Deformation-based morphometry provides a sensitive indicator of atrophy in Amyotrophic lateral sclerosis and has potential as a biomarker of disease burden, in both grey matter and white matter., (© The Author(s) (2020). Published by Oxford University Press on behalf of the Guarantors of Brain.)- Published
- 2020
- Full Text
- View/download PDF
13. Letter to the Editor: Comment on "The Impact of Chronic Liraglutide Therapy on Glucagon Secretion in Type 2 Diabetes: Insight From the LIBRA Trial" by Kramer C.K., et al.
- Author
-
Kalra S and Gupta Y
- Subjects
- Female, Humans, Male, Diabetes Mellitus, Type 2 blood, Diabetes Mellitus, Type 2 drug therapy, Glucagon blood, Hypoglycemic Agents therapeutic use, Liraglutide therapeutic use
- Published
- 2015
- Full Text
- View/download PDF
14. Efficacy and safety of 30-minute infusions of conivaptan in euvolemic and hypervolemic hyponatremia.
- Author
-
Koren MJ, Hamad A, Klasen S, Abeyratne A, McNutt BE, and Kalra S
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Benzazepines adverse effects, Double-Blind Method, Drug Administration Schedule, Female, Follow-Up Studies, Humans, Infusions, Intravenous, Least-Squares Analysis, Male, Middle Aged, Sodium blood, Time Factors, Young Adult, Antidiuretic Hormone Receptor Antagonists, Benzazepines therapeutic use, Body Water metabolism, Hyponatremia drug therapy
- Abstract
Purpose: The efficacy and safety of conivaptan administered via 30-minute i.v. infusion to patients with euvolemic or hypervolemic hyponatremia were evaluated., Methods: Hospitalized adults with a baseline serum sodium concentration (SSC) of 115-130 meq/L and euvolemia or hypervolemia on clinical evaluation were randomized to receive conivaptan hydrochloride 20 mg once or twice daily or placebo via 30-minute i.v. infusion. The primary efficacy measure was the change in SSC from baseline to 48 hours., Results: A total of 49 patients received one of the three treatment regimens. Conivaptan once and twice daily produced significant least-squares mean changes from baseline in SSC at 48 hours of 3.46 meq/L (95% confidence interval [CI], 1.75-5.18 meq/L) and 6.22 meq/L (95% CI, 4.34-8.10 meq/L), respectively (p = 0.028 between conivaptan-treated groups). These changes were significantly greater compared with those in the placebo group at hour 4 (p = 0.049) and at all time points onward of hour 28 (p ≤ 0.019) for the once-daily regimen and at all time points for the twice-daily regimen (p = 0.045 at hour 4, then p ≤ 0.010). Both conivaptan regimens were more efficacious than placebo in all secondary efficacy outcomes. Conivaptan was generally well tolerated, with infusion-site reactions being the most common adverse effects (AEs)., Conclusion: Conivaptan hydrochloride 20 mg, administered once or twice daily via 30-minute i.v. infusion, significantly increased SSCs over 48 hours in patients with euvolemic or hypervolemic hyponatremia when compared with placebo. Common AEs were similar to those seen with continuous conivaptan infusions.
- Published
- 2011
- Full Text
- View/download PDF
15. Effect of loading dose and formulation on safety and efficacy of conivaptan in treatment of euvolemic and hypervolemic hyponatremia.
- Author
-
Kalra S, Efrati S, Arthur JM, Oliven A, Velez JC, McNutt BE, Klasen S, and Abeyratne A
- Subjects
- Adult, Aged, Aged, 80 and over, Benzazepines therapeutic use, Blood Volume drug effects, Blood Volume physiology, Female, Follow-Up Studies, Humans, Hyponatremia physiopathology, Infusions, Intravenous, Male, Middle Aged, Treatment Outcome, Young Adult, Benzazepines administration & dosage, Benzazepines adverse effects, Chemistry, Pharmaceutical methods, Hyponatremia drug therapy
- Abstract
Purpose: The effect of loading dose and formulation on the safety and efficacy of conivaptan in the treatment of euvolemic and hypervolemic hyponatremia was studied., Methods: This parallel-group study randomized 121 hospitalized patients with euvolemic or hypervolemic hyponatremia to one of four treatment regimens: placebo loading dose followed by conivaptan continuous i.v. infusion using the ampul formulation (regimen 1), conivaptan loading dose followed by continuous i.v. infusion using the ampul formulation (regimen 2), placebo loading dose followed by conivaptan continuous i.v. infusion using the premixed formulation (regimen 3), or conivaptan loading dose followed by continuous i.v. infusion using the premixed formulation (regimen 4). The primary variable was the incidence and severity of injection-site reactions (ISRs), as evaluated using the ISR modified 5-point scale (ISRMS). Secondary outcomes included effects on serum sodium concentration (SSC), duration of effect, and safety and tolerability., Results: All four dosing regimens were efficacious, safe, and well tolerated. No significant differences in ISRMS scores or differences in changes from baseline SSC or in the duration of effects on SSC were observed between the regimens. Overly rapid SSC increases occurred in 7%, 7%, 3%, and 21% of patients treated with regimens 1, 2, 3, and 4, respectively. Overall, adverse events related to general disorders and ISRs occurred in 39%, 43%, 53%, and 55% of patients receiving regimens 1, 2, 3, and 4, respectively., Conclusion: Intravenous conivaptan regimens with or without a loading dose, whether using the ampul or a premixed formulation, had similar safety, tolerability, and efficacy in patients with euvolemic or hypervolemic hyponatremia. The pre-mixed formulation used with a loading dose may be associated with an increased frequency of overly rapid increase in SSC compared with the other regimens studied.
- Published
- 2011
- Full Text
- View/download PDF
16. Evidence for the existence of distinct central appetite, energy expenditure, and ghrelin stimulation pathways as revealed by hypothalamic site-specific leptin gene therapy.
- Author
-
Bagnasco M, Dube MG, Kalra PS, and Kalra SP
- Subjects
- Adipose Tissue, Brown chemistry, Animals, Arcuate Nucleus of Hypothalamus metabolism, Blood Glucose metabolism, Carrier Proteins genetics, Energy Intake, Fatty Acids, Nonesterified blood, Female, Gene Expression, Ghrelin, Green Fluorescent Proteins, Hypothalamus drug effects, In Situ Hybridization, Insulin blood, Ion Channels, Leptin blood, Luminescent Proteins genetics, Membrane Proteins genetics, Microinjections, Mitochondrial Proteins, Neuropeptide Y genetics, Paraventricular Hypothalamic Nucleus metabolism, Preoptic Area metabolism, Pro-Opiomelanocortin genetics, RNA, Messenger analysis, Rats, Rats, Sprague-Dawley, Reverse Transcriptase Polymerase Chain Reaction, Uncoupling Protein 1, Ventromedial Hypothalamic Nucleus metabolism, Weight Gain, Appetite physiology, Energy Metabolism, Hypothalamus metabolism, Leptin genetics, Peptide Hormones blood, Transfection
- Abstract
To identify the specific hypothalamic sites in which leptin acts to decrease energy intake and/or increase energy expenditure, recombinant adeno-associated virus vector-encoding leptin was microinjected bilaterally into one of four hypothalamic sites in female rats. Leptin transgene expression in the ventromedial nucleus and paraventricular nucleus induced comparable decreases in daily food intake (FI; 18-20%) and body weight (BW; 26-29%), accompanied by drastic reductions in serum leptin (81-97%), insulin (92-93%), free fatty acids (35-36%), and normoglycemia. Leptin transgene expression in the arcuate nucleus (ARC) decreased BW gain (21%) and FI (11%) to a lesser range, but the metabolic hormones were suppressed to the same extent. Leptin transgene expression in the medial preoptic area (MPOA) decreased BW and metabolic hormones without decreasing FI. Finally, leptin transgene expression in all four sites augmented serum ghrelin and thermogenic energy expenditure, as shown by uncoupling protein-1 mRNA expression in brown adipose tissue. Proopiomelanocortin gene expression in the ARC was up-regulated by leptin expression in all four sites, but neuropeptide Y gene expression in the ARC was suppressed by leptin transgene expression in the ARC but not in the MPOA. Thus, whereas leptin expression in the paraventricular nucleus, ventromedial nucleus, or ARC suppresses adiposity and insulin by decreasing energy intake and increasing energy expenditure, in the MPOA it suppresses these variables by increasing energy expenditure alone.
- Published
- 2002
- Full Text
- View/download PDF
17. Evidence that stimulation of two modalities of pituitary luteinizing hormone release in ovarian steroid-primed ovariectomized rats may involve neuropeptide Y Y1 and Y4 receptors.
- Author
-
Jain MR, Pu S, Kalra PS, and Kalra SP
- Subjects
- Amino Acid Sequence, Animals, Estradiol pharmacology, Female, Molecular Sequence Data, Neuropeptide Y antagonists & inhibitors, Neuropeptide Y pharmacology, Pancreatic Polypeptide pharmacology, Peptides, Cyclic pharmacology, Rats, Rats, Sprague-Dawley, Receptors, Neuropeptide Y agonists, Receptors, Neuropeptide Y antagonists & inhibitors, Estradiol analogs & derivatives, Luteinizing Hormone metabolism, Ovariectomy, Pituitary Gland metabolism, Progesterone pharmacology, Receptors, Neuropeptide Y physiology
- Abstract
A large body of evidence indicates that neuropeptide Y (NPY) is involved in stimulation of basal and cyclic release of hypothalamic LHRH and pituitary LH. To identify the NPY receptor subtypes that mediate the excitatory effects of NPY in these two modalities of LH release, we studied the effects of 1229U91, a selective Y1 receptor antagonist and Y4 receptor agonist, in two experimental paradigms that reproduce the two modalities of LH secretion in steroid-primed ovariectomized (OVX) rats. Rats were ovariectomized and implanted with a permanent cannula into the lateral cerebroventricle. In the first experiment, rats received estradiol benzoate (EB, 30 microg/rat) on day 5, followed 2 days later with progesterone (2 mg/rat) at 1000 h to induce an afternoon LH surge. 1229U91 (30 microg/3 microl) or vehicle (control) was injected intracerebroventricularly into these rats either once at 1300 h or twice (15 microg/injection) at 1100 and 1200 h. Blood samples were collected before progesterone injection at 1000 h and at hourly intervals from 1300 -1800 h via an intrajugular cannula implanted on the previous day. In control rats, serum LH levels rose significantly at 1400 h, and these high levels were maintained until 1700 h. After two injections of 1229U91, LH levels displayed a tendency to rise at 1300-1400 h, as in controls, but thereafter, decreased rapidly below the control range. In the second experiment, the acute effect of 1229U91 on LH release was evaluated in OVX rats pretreated with EB alone. Saline alone or saline containing 1, 3, 10, or 30 microg 1229U91 was injected intracerebroventricularly at 1000 h, and the effects on LH release were analyzed at 10, 20, 30, and 60 min. 1229U91 elicited a dose-dependent stimulation of LH release, with maximal response (950% of basal levels) occurring at 10 min after the 30-microg dose; elevated levels were maintained for 1 h. Because 1229U91 is a potent Y4 agonist with some affinity for Y5 receptors, these results raised the possibility that activation of Y4/Y5 receptors by 1229U91 may augment LH release. Therefore, we examined the effects of icv administration of rat pancreatic polypeptide, a Y4-selective agonist, and [D-Trp32]-NPY, a Y5 agonist on LH release in EB-primed rats. Rat pancreatic polypeptide (0.5-2 microg/rat) stimulated LH release in a dose-related manner, and peak levels (280% of basal levels) were seen at 10-20 min; the response evoked by a higher dose (10 microg) was smaller than that induced by 0.5 or 2 microg. [D-Trp32]-NPY was relatively less effective, because only the highest (10-microg) dose elicited a modest stimulation (244% of basal levels). These results demonstrate that 1229U91, a Y1 antagonist and Y4 agonist, evokes two types of responses; it suppresses the protracted ovarian steroid-induced LH surge, and acutely, it also stimulates LH. These results imply that normally two different types of NPY receptors may mediate the stimulation of LH release. Because 1229U91 is a Y1 receptor antagonist, inhibition of the steroid-induced LH surge by 1229U91 suggests that Y1 receptors may mediate the cyclic release of LH. On the other hand, acute stimulation of LH by 1229U91 implies that the Y4 agonist-like activity of 1229U91 may mediate the basal release of LH and that either NPY or a yet-to-be-identified endogenous Y4 receptor agonist may activate Y4 receptors in the hypothalamus to stimulate LH release.
- Published
- 1999
- Full Text
- View/download PDF
18. Inhibition of neuropeptide Y (NPY)-induced feeding and c-Fos response in magnocellular paraventricular nucleus by a NPY receptor antagonist: a site of NPY action.
- Author
-
Yokosuka M, Kalra PS, and Kalra SP
- Subjects
- Animals, Eating drug effects, Immunohistochemistry, Male, Neuropeptide Y antagonists & inhibitors, Neuropeptide Y pharmacology, Paraventricular Hypothalamic Nucleus drug effects, Peptides, Cyclic pharmacology, Proto-Oncogene Proteins c-fos antagonists & inhibitors, Rats, Rats, Sprague-Dawley, Receptors, Neuropeptide Y antagonists & inhibitors, Eating physiology, Neuropeptide Y physiology, Paraventricular Hypothalamic Nucleus physiology, Proto-Oncogene Proteins c-fos metabolism, Receptors, Neuropeptide Y physiology
- Abstract
Neuropeptide Y (NPY) is one of the important endogenous orexigenic peptides. In these studies we employed c-Fos immunostaining and a selective NPY Y1 receptor antagonist to identify the site of action of NPY in the hypothalamus. The results showed that intracerebroventricular administration of NPY stimulated feeding and increased immunostaining of c-Fos, a product of the immediate early gene c-fos, in several hypothalamic sites, including the dorsomedial nucleus, the supraoptic nucleus, and the two subdivisions of the paraventricular nucleus (PVN), the parvocellular PVN, and magnocellular PVN (mPVN). Intracerebroventricular administration of 1229U91, a selective NPY Y1 receptor antagonist, affected neither food intake nor c-Fos-like immunoreactivity (FLI) in these hypothalamic sites. Co-administration of NPY and NPY Y1 receptor antagonist inhibited NPY-induced food intake by 48%, but failed to affect NPY-induced FLI in the supraoptic nucleus, dorsomedial nucleus, and parvocellular PVN. However, this combined treatment decreased FLI by 46% in the mPVN (P < 0.05). These results showed that whereas NPY can stimulate FLI in several hypothalamic sites, the selective NPY Y1 antagonist suppressed NPY-induced FLI only in the mPVN. Thus, these findings lend credence to the view that a subpopulation of Y1 receptor-containing neurons in the mPVN in part mediate stimulation of feeding by NPY.
- Published
- 1999
- Full Text
- View/download PDF
19. Daily changes in hypothalamic gene expression of neuropeptide Y, galanin, proopiomelanocortin, and adipocyte leptin gene expression and secretion: effects of food restriction.
- Author
-
Xu B, Kalra PS, Farmerie WG, and Kalra SP
- Subjects
- Animals, Body Weight, Carrier Proteins analysis, Corticosterone blood, Leptin, Male, Proteins metabolism, Rats, Rats, Sprague-Dawley, Receptors, Leptin, Adipocytes metabolism, Eating, Galanin genetics, Gene Expression, Hypothalamus metabolism, Neuropeptide Y genetics, Pro-Opiomelanocortin genetics, Proteins genetics, Receptors, Cell Surface
- Abstract
The participation of hypothalamic neuropeptide Y (NPY)-, galanin (GAL)-, and opioid-producing neurons in the restraint on food intake exerted by adipocyte leptin has recently been recognized. To further understand the interplay between the central appetite-stimulating- and peripheral appetite-inhibiting signals in the management of daily food intake, we have examined the daily patterns in expression of the hypothalamic neuropeptides and leptin receptor (R) and adipocyte leptin gene expression and secretion in freely feeding (FF) rats. These analyses were extended to determine the impact of food restriction (FR) to 4 h daily for 4 weeks. Groups of FF and FR rats were killed at 4-h intervals during a 24-h period, and hypothalamic NPY, GAL, POMC, and leptin-R gene expression and leptin gene expression were evaluated by RNase protection assays and serum leptin and corticosterone (CORT) levels were estimated by RIA. The following new findings emerged: 1) In FF rats, hypothalamic NPY messenger RNA (mRNA) levels fluctuated during the course of 24 h with high levels at 0700 h and 1100 h followed by a decrease at 1500 h during the lights-on phase that was sustained throughout the dark phase (1900 h-0500 h) of the light-dark cycle. Hypothalamic GAL and POMC mRNA also displayed daily patterns but with a different time course; GAL and POMC gene expression were elevated 4 h later than NPY mRNA at 1100 h and 1500 h. 2) Although FR to 4 h between 1100 h and 1500 h resulted in maintenance of body weight compared with a steady weight gain in FF rats, the daily patterns of fluctuations in hypothalamic neuropeptide gene expression were abolished. 3) In FF rats, hypothalamic leptin-R and adipocyte leptin gene expression and serum leptin levels displayed a daily pattern temporally different from that of hypothalamic neuropeptide gene expression. Adipocyte leptin mRNA remained low during the lights-on phase but increased at the onset of the lights-off phase (1900 h) and remained elevated through the dark phase. 4) Hypothalamic leptin-R gene expression, like that of adipocyte leptin gene expression, rose abruptly at the onset of nocturnal feeding behavior but receded progressively to low range thereafter. 5) On the other hand, a dichotomy in the daily rise in adipocyte leptin gene expression and leptin secretion was observed in FF rats. Unlike adipocyte leptin mRNA, serum leptin increased at 2300 h, 4 h after initiation of ingestive behavior. 6) In FR rats, adipocyte leptin gene expression fluctuated little over the 24-h period but, as in FF rats, leptin hypersecretion peaked 4 h after initiation of food intake. 7) In both FF and FR rats, increased serum CORT levels preceded serum leptin rise. Overall, these results show that in FF rats, gene expression of hypothalamic appetite stimulating peptides first rise and then fall to nadir during the lights-on phase when leptin levels are in low range; adipocyte leptin mRNA rises before impending ingestive behavior and increased leptin secretion reaching peak manifests itself during nocturnal feeding. The FR regimen, which curtailed the normal body weight gain, abolished these daily fluctuations in gene expression of hypothalamic orexigenic peptides and adipocyte leptin but permitted feeding-associated increased leptin secretion. Thus, it may be important to consider the daily patterns of gene expression and availability of hypothalamic orexigenic peptides in investigations aimed at elucidating the central mechanisms underlying the feedback action of the normal and altered leptin secretion patterns.
- Published
- 1999
- Full Text
- View/download PDF
20. Interactions between neuropeptide Y and gamma-aminobutyric acid in stimulation of feeding: a morphological and pharmacological analysis.
- Author
-
Pu S, Jain MR, Horvath TL, Diano S, Kalra PS, and Kalra SP
- Subjects
- Animals, Drug Synergism, GABA Agonists pharmacology, Male, Muscimol pharmacology, Neuropeptide Y metabolism, Paraventricular Hypothalamic Nucleus metabolism, Rats, Rats, Sprague-Dawley, Tissue Distribution, gamma-Aminobutyric Acid metabolism, Eating drug effects, Neuropeptide Y pharmacology, gamma-Aminobutyric Acid pharmacology
- Abstract
Neuropeptide Y (NPY) produced in neurons in the arcuate nucleus and brain stem and released in the paraventricular nucleus (PVN) and surrounding areas is involved in stimulation of feeding in rats. We recently reported that gamma-aminobutyric acid (GABA) is coexpressed in a subpopulation of NPY neurons in the arcuate nucleus. To determine whether GABA is colocalized in NPY terminals in the PVN, the site of NPY action, light and electron microscopic double staining for NPY and GABA using pre- and postembedding immunolabeling was performed on rat brain sections. GABA was detected in NPY-immunopositive axons and axon terminals within both the parvocellular and magnocellular divisions of the PVN. These morphological findings suggested a NPY-GABA interaction in the hypothalamic control of feeding. Therefore, the effects of muscimol (MUS), a GABA(A) receptor agonist, on NPY-induced food intake were examined in sated rats. When injected intracerebroventricularly, both NPY and MUS elicited dose-dependent feeding responses that were blocked by the administration of 1229U91 (a putative Y1 receptor antagonist) or bicuculline (a GABA(A) receptor antagonist), respectively. Coadministration of NPY and MUS intracerebroventricularly amplified the feeding response over that evoked by NPY or MUS alone. Similarly, microinjection of either NPY or MUS into the PVN stimulated food intake in a dose-related fashion, and coinjection elicited a significantly higher response than that evoked by either individual treatment. These results suggest that GABA and NPY may coact through distinct receptors and second messenger systems in the PVN to augment food intake.
- Published
- 1999
- Full Text
- View/download PDF
21. Interacting appetite-regulating pathways in the hypothalamic regulation of body weight.
- Author
-
Kalra SP, Dube MG, Pu S, Xu B, Horvath TL, and Kalra PS
- Subjects
- Animals, Humans, Leptin, Peptides physiology, Proteins physiology, Appetite Regulation physiology, Body Weight, Homeostasis, Hypothalamus physiology
- Abstract
Various aspects of the complex spatio-temporal patterning of hypothalamic signaling that leads to the development of synchronized nocturnal feeding in the rat are critically examined. Undoubtedly, as depicted in Fig. 7, a distinct ARN in the hypothalamus is involved in the control of nocturnal appetite. At least four basic elements operate within this ARN. These are: 1) A discrete appetite-driving or orexigenic network of NPY, NE, GABA, GAL, EOP, and orexin transduces and releases appetite-stimulating signals. 2) Similarly, anorexigenic signal-producing pathways (e.g., CRH, GLP-1, alpha MSH, and CART) orchestrate neural events for dissipation of appetite and to terminate feeding, possibly by interrupting NPY efflux and action at a postsynaptic level within the hypothalamus. It is possible that some of these may represent the physiologically relevant "off" switches under the influence of GABA alone, or AgrP alone, or in combination with NPY released from the NPY-, GABA-, and AgrP-coproducing neurons. 3) Recent evidence shows that neural elements in the VMN-DMN complex tonically restrain the orexigenic signals during the intermeal interval; the restraint is greatly aided by leptin's action via diminution of orexigenic (NPY) and augmentation of anorexigenic (GLP-1, alpha MSH, and CART) signals. Since interruption of neurotransmission in the VMN resulted in hyperphagia and development of leptin resistance, it seems likely that the VMN is an effector site for the restraint exercised by leptin. The daily rhythms in leptin synthesis and release are temporally dissociable because the onset of daily rise in leptin gene expression in adipocytes precedes that in leptin secretion. Nevertheless, these rhythms are in phase with daily ingestive behavior because the peak in circulating leptin levels occurs during the middle of the feeding period. These observations, coupled with the fact that circulating levels of leptin are directly related to adiposity, pose a new challenge for elucidating the precise role of leptin in daily patterning of feeding in the rat. 4) A neural timing mechanism also operates upstream from the ARN in the daily management of energy homeostasis. Although the precise anatomical boundaries are not clearly defined, this device is likely to be composed of a group of neurons that integrate incoming internal and external information for the timely onset of the drive to eat. Evidently, this network operates independently in primates, but it is entrained to the circadian time keeper in the SCN of rodents. Apart from its role in the onset of drive to eat, the circadian patterns of gene expression of NPY, GAL, and POMC denote independent control of the timing device on the synthesis and availability for release of orexigenic signals. The VMN-DMN-PVN complex is apparently an integrated constituent of the timing mechanism in this context, because lesions in each of these sites result in loss of regulated feeding. The accumulated evidence points to the PVN and surrounding neural sites within this framework as the primary sites of release and action of various orexigenic and anorexigenic signals. A novel finding is the identification of the interconnected wiring of the DMN-mPVN axis that may mediate leptin restraint on NPY-induced feeding. The chemical phenotypes of leptin and NPY target neurons in this axis remain to be identified. These multiple orexigenic and anorexigenic pathways in the hypothalamic ARN appear to represent redundancy, a characteristic of regulated biological systems to provide a "fail-safe" neural mechanism to meet an organism's constant energy needs for growth and maintenance. Within this formulation, the coexisting orexigenic signals (NPY, NE, GAL, GABA, and AgrP) represent either another level of redundancy or it is possible that these signals operate within the ARN as reinforcing agents to varying degrees under different circumstances. (ABSTRACT TRUNCATED)
- Published
- 1999
- Full Text
- View/download PDF
22. Absence of increased neuropeptide Y neuronal activity before and during the luteinizing hormone (LH) surge may underlie the attenuated preovulatory LH surge in middle-aged rats.
- Author
-
Sahu A and Kalra SP
- Subjects
- Animals, Female, Gene Expression physiology, Hypothalamus cytology, Hypothalamus metabolism, Neuropeptide Y genetics, Proestrus metabolism, Rats, Tissue Distribution, Aging metabolism, Follicular Phase blood, Luteinizing Hormone blood, Neurons metabolism, Neuropeptide Y metabolism
- Abstract
A large body of evidence suggests that the neuroendocrine axis plays a major role in the reproductive aging of female rats. Since increased hypothalamic neuropeptide Y (NPY) neurosecretion is crucial in the preovulatory LH discharge in young rats, we tested the hypothesis that diminution in the preovulatory LH surge in middle-aged (MA) rats may be due to altered neurosecretory activity in NPYergic neurons. In Exp 1, we examined NPY levels in six microdissected hypothalamic nuclei, including median eminence (ME), arcuate nucleus (ARC), and medial preoptic area (MPOA), at 1000, 1200, 1400, 1600, 1800, 2000, or 2200 h on the day of proestrus in young (2.5- to 3-month old) and MA (7- to 9-month old) regularly cycling rats. At 1000 h, ME NPY levels in young rats were significantly lower than those in MA rats. In young rats, the ME NPY levels were significantly increased at 1400 h before the LH surge in the afternoon and thereafter decreased progressively during the interval of the LH surge. In MA rats, however, ME NPY levels decreased in the afternoon in association with an attenuated LH surge. In addition, in the ARC and MPOA, the other hypothalamic sites associated with induction of LH surge, NPY levels increased before and during the LH surge in young rats, no change in NPY levels in these nuclei was observed in association with the attenuated LH surge in MA rats. Also, NPY levels in the ARC and MPOA during the afternoon were significantly lower in MA compared with those in young animals. These results demonstrated the absence of an antecedent increase in NPY levels, specifically in the ME and ARC, during the afternoon of proestrus in MA animals. In a second experiment, we evaluated whether the absence of dynamic changes in NPY levels in the ME and ARC in MA rats was due to altered hypothalamic NPY gene expression. Regularly cycling young (2.5- to 3-month-old) and MA (8- to 10-month-old) rats were killed at 1000, 1200, 1400, 1600, 1800, 2000, or 2200 h on the day of proestrus. The medial basal hypothalamus was processed for prepro-NPY messenger RNA (mRNA) measurement by ribonuclease protection assay. In young rats, prepro-NPY mRNA levels were significantly increased at 1200 h and remained elevated throughout the afternoon. In contrast, in MA rats prepro-NPY mRNA levels remained unchanged before and during the attenuated LH surge. These results clearly indicate that the augmentation in NPY neuronal activity before and during the LH surge seen in young rats fails to manifest itself in middle-aged rats. As hypothalamic NPY participates in the induction of LHRH surge, our results suggest that reduced LHRH and LH surges in MA rats may be due to diminution in NPY secretion in these animals.
- Published
- 1998
- Full Text
- View/download PDF
23. Anorectic effects of the cytokine, ciliary neurotropic factor, are mediated by hypothalamic neuropeptide Y: comparison with leptin.
- Author
-
Xu B, Dube MG, Kalra PS, Farmerie WG, Kaibara A, Moldawer LL, Martin D, and Kalra SP
- Subjects
- Adipocytes drug effects, Adipocytes physiology, Animals, Body Weight drug effects, Ciliary Neurotrophic Factor, Eating drug effects, Gene Expression drug effects, Hypothalamus drug effects, Hypothalamus physiology, Injections, Intraperitoneal, Injections, Intraventricular, Leptin, Male, Neuropeptide Y genetics, Neuropeptide Y pharmacology, Proteins genetics, Rats, Rats, Sprague-Dawley, Anorexia chemically induced, Cytokines, Hypothalamus metabolism, Nerve Tissue Proteins pharmacology, Neuropeptide Y physiology, Proteins pharmacology
- Abstract
Although ciliary neurotropic factor (CNTF) is a tropic factor in nervous system development and maintenance, peripheral administration of this cytokine also causes severe anorexia and weight loss. The neural mechanism(s) mediating the loss of appetite is not known. As hypothalamic neuropeptide Y (NPY) is a potent orexigenic signal, we tested the hypothesis that CNTF may adversely affect NPYergic signaling in the hypothalamus. Intraperitoneal administration of CNTF (250 microg/kg) daily for 4 days significantly suppressed 24-h food intake in a time-dependent manner and decreased body weight. The loss in body weight was similar to that which occurred in pair-fed (PF) rats. As expected, hypothalamic NPY gene expression, determined by measurement of steady state prepro-NPY messenger RNA by ribonuclease protection assay, significantly increased in PF rats in response to energy imbalance. However, despite a similar loss in body weight, there was no increase in NPY gene expression in CNTF-treated rats. Daily administration of CNTF intracerebroventricularly (0.5 or 5.0 microg/rat) also produced anorexia and body weight loss. In this experiment, negative energy balance produced by both PF and food deprivation augmented hypothalamic NPY gene expression. However, despite reduced intake and loss of body weight, no similar increment in hypothalamic NPY gene expression was observed in CNTF-treated rats. In fact, in rats treated with higher doses of CNTF (5.0 microg/rat), NPY gene expression was reduced below the levels seen in control, freely fed rats. Furthermore, CNTF treatment also markedly decreased NPY-induced feeding. These results suggested that anorexia in CNTF-treated rats may be due to a deficit in NPY supply and possibly in the release and suppression of NPY-induced feeding. The possibility that CNTF-induced anorexia may be caused by increased leptin was next examined. Daily intracerebroventricular injections of leptin (7 microg/rat) decreased food intake, body weight, and hypothalamic NPY gene expression in a manner similar to that seen after CNTF treatment. Leptin administration also suppressed NPY-induced feeding. However, peripheral and central CNTF injections markedly decreased leptin messenger RNA in lipocytes, indicating a deficiency of leptin in these rats; thus, leptin was unlikely to be involved in appetite suppression. Thus, these results show that a two-pronged central action of CNTF, causing diminution in both NPY availability and the NPY-induced feeding response, may underlie the severe anorexia. Further, unlike other members of the cytokine family, suppression of NPYergic signaling in the hypothalamus by CNTF does not involve up-regulation of leptin, but may involve a direct action on hypothalamic NPY neurons or on neural circuits that regulate NPY signaling in the hypothalamus.
- Published
- 1998
- Full Text
- View/download PDF
24. Evidence showing that beta-endorphin regulates cyclic guanosine 3',5'-monophosphate (cGMP) efflux: anatomical and functional support for an interaction between opiates and nitric oxide.
- Author
-
Pu S, Horvath TL, Diano S, Naftolin F, Kalra PS, and Kalra SP
- Subjects
- Animals, Microscopy, Electron, Morphine pharmacology, Nitric Oxide Synthase metabolism, Preoptic Area metabolism, Rats, Cyclic GMP metabolism, Naloxone pharmacology, Nitric Oxide metabolism, Preoptic Area drug effects, beta-Endorphin physiology
- Abstract
Nitric oxide (NO) is now recognized as a diffusible messenger molecule that normally augments intercellular communication in the central nervous system, but is neurotoxic if released in excessive amounts. NO is synthesized from L-arginine by the Ca2+/calmodulin-dependent neuronal isoform NO synthase (NOS) localized in sub-populations of neurons throughout the brain, including the hypothalamus. In the hypothalamus, NO stimulates the release of GnRH, the primary neurohormone governing reproduction in mammals. Although the excitatory amino acid, glutamate, acting through the N-methyl-D-aspartate (NMDA) receptor is believed to be responsible for stimulation of NO release, the neuronal system(s) that inhibits NO efflux is unknown. As the endogenous opioids, primarily beta-endorphin (betaEND), exert a tonic restraint on GnRH secretion, we sought evidence for a possible functional link between betaEND and NOS pathways in the hypothalamus. We observed that restraining the opioid influence with the opiate receptor antagonist, naloxone, in intact, but not in castrated, rats rapidly augmented extracellular cGMP/NO efflux in the medial preoptic area, where GnRH, NOS, and betaEND immunoreactive pathways are coextensive. Pituitary LH secretion increased in conjunction with this augmented cGMP/NO response and pretreatment with the mu opiate receptor agonist, morphine, suppressed these naloxone-induced responses. Further, visualization of hypothalamic sections immunostained for both betaEND and NOS revealed betaEND-immunoreactive axon terminals in close proximity to NOS-positive cell bodies and dendrites in a number of hypothalamic subdivisions, including the medial preoptic area. These close appositions represented conventional synapses between betaEND nerve terminals and NOS-positive perikarya and dendrites under the electron microscope. Clearly, the experimental data, corroborated by morphological evidence, point to a direct inhibitory control of betaEND on NOS-immunoreactive neurons in monitoring cGMP/NO release. These findings together with the previous observations that the glutamate neurotransmitter acting through NMDA receptors located on NOS-immunopositive cells stimulates cGMP/NO efflux and plasma LH selectively in intact rats document the existence of a dual control comprised of the excitatory NMDA and the inhibitory mu opiate receptors in modulating cGMP/NO release, a response also directed by gonadal steroids. This new knowledge of an inhibitory opioid influence on cGMP/NO release is probably extremely important both in the generation of periodicities in GnRH secretion that underlie hypothalamic control of reproduction and in protecting against neurotoxic overstimulation of NO release by excitatory amino acids.
- Published
- 1997
- Full Text
- View/download PDF
25. An interactive physiological role of neuropeptide Y and galanin in pulsatile pituitary luteinizing hormone secretion.
- Author
-
Xu B, Pu S, Kalra PS, Hyde JF, Crowley WR, and Kalra SP
- Subjects
- Animals, Antibodies immunology, Female, Galanin genetics, Galanin immunology, Neuropeptide Y genetics, Neuropeptide Y immunology, Oligonucleotides, Antisense pharmacology, Ovariectomy, Pulsatile Flow, Rats, Rats, Sprague-Dawley, Galanin physiology, Luteinizing Hormone metabolism, Neuropeptide Y physiology, Pituitary Gland metabolism
- Abstract
Both neuropeptide Y (NPY) and galanin (GAL) systems have been implicated in the excitatory regulation of pulsatile LH secretion in the ovariectomized rat. The present studies were designed to examine the possible interaction of these two neuropeptides in controlling episodic LH release by testing the effects of central infusion of antibodies (Ab) to NPY and GAL, alone or in combination; additional studies tested the effects of central administration of an antisense oligodeoxynucleotide (ODN) to NPY and GAL messenger RNA. Rats were ovariectomized, implanted with a cannula in the third ventricle, and used in experiments 2 weeks later. Central infusion, via Alzet osmotic minipumps, of IgG purified from an NPY Ab produced a dose-related suppression of pulsatile LH secretion. Although an Ab dilution of 1:10 was ineffective, a maximal inhibitory effect was obtained using an NPY Ab dilution of 1:1, which decreased the mean levels, pulse frequency, and pulse amplitude of LH. These parameters of episodic LH secretion were also significantly reduced by central injection of antisense NPY ODN compared to those in vehicle- or missense ODN-treated controls. Similar dose-related inhibitory effects on the parameters of LH secretion were seen after central infusion of GAL Ab. Furthermore, infusion of a combination of NPY Ab and GAL Ab, each at the ineffective dilution of 1:10, resulted in a profound inhibition of LH secretion equivalent to the pattern seen with the maximally effective 1:1 Ab dilution. These results strengthen the idea of a physiological role for both NPY and GAL systems in the mechanism underlying the LHRH pulse generator activity and further suggest that these two excitatory neuropeptides act in concert to generate pulsatile LHRH release.
- Published
- 1996
- Full Text
- View/download PDF
26. Neuropeptide Y Y2 receptors in hypothalamic neuroendocrine areas are up-regulated by estradiol and decreased by progesterone cotreatment in the ovariectomized rat.
- Author
-
Parker SL, Carroll BL, Kalra SP, St-Pierre S, Fournier A, and Crowley WR
- Subjects
- Analysis of Variance, Animals, Drug Interactions, Female, Humans, Hypothalamus drug effects, Hypothalamus, Anterior metabolism, Hypothalamus, Middle metabolism, Luteinizing Hormone blood, Organ Specificity, Ovariectomy, Peptide YY, Point Mutation, Preoptic Area metabolism, Prolactin blood, Rats, Recombinant Proteins metabolism, Down-Regulation, Estradiol pharmacology, Hypothalamus metabolism, Peptides metabolism, Progesterone pharmacology, Receptors, Neuropeptide Y biosynthesis, Up-Regulation
- Abstract
Central neuropeptide Y (NPY) systems are known to stimulate, via the Y1 subtype of NPY receptor, the release of LHRH that leads to surges of LH. Levels of NPY receptors in relation to the above modulation have not been assessed. This study, therefore, examined profiles of NPY receptors in ovariectomized (Ovx) rats and in Ovx rat treated with estradiol (E2) with or without cotreatment with progesterone (P4). [125I]Human PYY containing proline in position 34 [(Pro 34 )hPYY] was used as the Y1 receptor ligand, and [125 I]human peptide YY-(3-36) [hPYY-(3-36)] was used as the Y2 site ligand. Treatment with E2 over 3 days increased Y2 binding in the preoptic hypothalamus and the medial basal hypothalamus, whereas no changes were found in the lateral anterior hypothalamus or the piriform cortex. Administration of P4 (1.5 mg/animal) on the third day of E2 treatment reduced Y2 binding in both the preoptic hypothalamus and the medial basal hypothalamus to or below the density found in Ovx controls. The Y1 receptor levels and the affinity of either Y1 or Y2 binding did not change appreciably with any of the treatments. No significant changes in the binding of wheat germ agglutinin were found at the time of the largest reduction in Y2 receptor numbers by P4, indicating the absence of a major membrane receptor reduction in response to the progestin. The down-regulation of Y2 sites by P4 preceded and accompanied the surge of serum LH induced by the progestin in E2-treated animals. A short term P4 treatment thus appears to reduce the Y2 tone in hypothalamic areas involved in LHRH secretion. This reduction might reinforce Y1 drives known to stimulate the output of LHRH, and thus contribute to LH release.
- Published
- 1996
- Full Text
- View/download PDF
27. Morphological and pharmacological evidence for neuropeptide Y-galanin interaction in the rat hypothalamus.
- Author
-
Horvath TL, Naftolin F, Leranth C, Sahu A, and Kalra SP
- Subjects
- Analysis of Variance, Animals, Arcuate Nucleus of Hypothalamus cytology, Arcuate Nucleus of Hypothalamus physiology, Axons physiology, Axons ultrastructure, Cerebral Ventricles drug effects, Cerebral Ventricles physiology, Colchicine pharmacology, Dendrites physiology, Dendrites ultrastructure, Estradiol pharmacology, Estrus, Female, Galanin analogs & derivatives, Galanin analysis, Galanin pharmacology, Hypothalamus cytology, Hypothalamus drug effects, Hypothalamus, Anterior cytology, Hypothalamus, Anterior physiology, Immunoenzyme Techniques, Immunohistochemistry, Luteinizing Hormone metabolism, Microscopy, Immunoelectron, Neurons cytology, Neuropeptide Y analysis, Ovariectomy, Paraventricular Hypothalamic Nucleus cytology, Paraventricular Hypothalamic Nucleus physiology, Preoptic Area cytology, Preoptic Area physiology, Progesterone pharmacology, Rats, Rats, Sprague-Dawley, Signal Transduction, Substance P analogs & derivatives, Substance P pharmacology, Supraoptic Nucleus cytology, Supraoptic Nucleus physiology, Galanin physiology, Hypothalamus physiology, Neurons physiology, Neuropeptide Y physiology
- Abstract
Galanin (GAL) and neuropeptide Y (NPY) have been shown to play important roles in the regulation of pituitary hormone secretion, as well as ingestive and sexual behaviors, by acting within the hypothalamus. While the mechanism of action of these regulatory peptides is under intensive investigation, less attention has been paid to the possible interaction between them in influencing these central regulatory processes. Because NPY and GAL augment pituitary gonadotropin release, the present study was undertaken to evaluate the nature of morphological and functional relationships between these excitatory hypothalamic peptidergic systems. Double immunolabeling for NPY and GAL was carried out on vibratome sections taken from the hypothalamus of colchicine-pretreated female rats. Avidinbiotin peroxidase technique and a dark blue diaminobenzidine reaction was used to visualize NPY profiles, while the GAL neurons were labeled with a light brown diaminobenzidine reaction using either the avidin-biotin peroxidase or the peroxidase antiperoxidase technique. Light microscopic examination of the immunostained material showed that in the arcuate nucleus, paraventricular nucleus, supraoptic nucleus, anterior hypothalamus, and medial preoptic area, an abundant network of NPY-immunoreactive axons surrounded GAL-immunostained cells. Numerous dark blue NPY-containing putative boutons were observed in close proximity to GAL-immunolabeled cell bodies and dendrites. Correlated light and electron microscopic examination revealed that most of the immunoreactive NPY axon terminals established synaptic connections with GAL-expressing cells. Synaptic connections were most frequently found in the medial preoptic area and in the magnocellular region of the paraventricular nucleus and arcuate nucleus. Fewer connections were observed in the supraoptic nucleus. These morphological observations demonstrate the existence of a strong NPY input to hypothalamic GAL neurons, thereby suggesting a modulatory role for NPY in monitoring GAL release. To evaluate the functional relevance of this anatomical relationship, the effects of intraventricular injection of a GAL receptor antagonist, galantide, were examined on NPY-induced LH release in ovarian steroid-primed ovariectomized rats. As expected, intraventricular injection of NPY readily stimulated LH release. Although, while on its own, galantide was ineffective in altering basal LH release, it markedly attenuated the NPY-induced LH response, thereby suggesting that GAL released in response to NPY administration may, in part, mediate the excitatory effects of NPY. These experimental results, taken together with the morphological observations, document the involvement of an NPY --> GAL signaling modality in the release of gonadotropins and, likewise, raise the possibility of a similar signaling process in the release of other pituitary hormones and elicitation of behavioral effects attributed to NPY and GAL.
- Published
- 1996
- Full Text
- View/download PDF
28. Evidence that gonadal steroids modulate nitric oxide efflux in the medial preoptic area: effects of N-methyl-D-aspartate and correlation with luteinizing hormone secretion.
- Author
-
Pu S, Xu B, Kalra SP, and Kalra PS
- Subjects
- 2-Amino-5-phosphonovalerate pharmacology, Animals, Arginine analogs & derivatives, Arginine pharmacology, Cyclic GMP metabolism, Enzyme Inhibitors pharmacology, Female, Luteinizing Hormone blood, Male, NG-Nitroarginine Methyl Ester, Nitric Oxide Synthase antagonists & inhibitors, Rats, Rats, Sprague-Dawley, Receptors, N-Methyl-D-Aspartate antagonists & inhibitors, Luteinizing Hormone metabolism, N-Methylaspartate pharmacology, Nitric Oxide metabolism, Preoptic Area drug effects, Preoptic Area metabolism
- Abstract
Several lines of evidence suggest that nitric oxide (NO) is involved in the neuroendocrine control of reproductive function. This study was undertaken to determine 1) NO activity in the medial preoptic area (MPOA) where LHRH- and NO synthase-containing neurons are coextensive; 2) whether N-methyl-D-aspartate (NMDA) receptor activation, which stimulates LHRH release, augments NO activity in the MPOA; and 3) whether NO activation in the MPOA underlies the steroid dependency of NMDA-induced pituitary LH release. As extracellular levels of cGMP in discrete brain sites are a reliable index of basal and stimulated activity of NO, extracellular cGMP levels in the MPOA of freely moving, awake rats were measured by microdialysis in the current study. In the first experiment, the MPOA of intact and castrated male rats were microdialyzed with artificial cerebrospinal fluid at a rate of 5 microliters/min. The basal level of cGMP efflux was determined from the initial seven samples collected at 20-min intervals. The NO response to a single i.v. injection of NMDA (10 mg/kg) or saline was assessed in the next five samples. In the second experiment, the basal and NMDA-evoked NO effluxes in the MPOA of ovariectomized (ovx) and estrogen-treated ovx rats were examined. Results showed that in both sexes, the absence of gonadal steroids resulted in significantly lower basal cGMP levels. Additionally, the cGMP response to NMDA was steroid dependent. Whereas in castrated rats it failed to affect cGMP efflux, NMDA in intact male rats promptly raised cGMP levels at 20 min, and these elevated levels were maintained through the duration of the experiment. This NMDA-induced cGMP response, observed selectively in intact rats, was also associated with stimulation of plasma LH levels. In female rats, NMDA similarly enhanced MPOA cGMP efflux and pituitary LH secretion in estradiol benzoate-treated, but not in oil-treated, ovx rats. The NMDA receptor antagonist D,L-amino-5-phosphoropentanoic acid and the NO synthase inhibitor, N omega-nitro-L-arginine, completely blocked the NMDA-induced cGMP and LH responses, thereby demonstrating the specificity of the NMDA receptor --> NO line of communication probably operating in the MPOA in the control of pituitary LH release. Therefore, these results show that gonadal steroids augment basal as well as NMDA-induced MPOA cGMP efflux in male and female rats. It is likely that facilitation of NO/cGMP activity in the MPOA may underlie the steroid dependency of NMDA-evoked LH hypersecretion in the rat.
- Published
- 1996
- Full Text
- View/download PDF
29. L-arginine/nitric oxide amplifies the magnitude and duration of the luteinizing hormone surge induced by estrogen: involvement of neuropeptide Y.
- Author
-
Bonavera JJ, Kalra PS, and Kalra SP
- Subjects
- Animals, Base Sequence, Female, Gonadotropin-Releasing Hormone metabolism, Hypothalamus, Middle drug effects, Hypothalamus, Middle metabolism, Molecular Sequence Data, Neuropeptide Y genetics, Neuropeptide Y metabolism, Oligonucleotides, Antisense pharmacology, Ovariectomy, Preoptic Area drug effects, Preoptic Area metabolism, Rats, Arginine pharmacology, Estradiol pharmacology, Luteinizing Hormone metabolism, Neuropeptide Y physiology, Nitric Oxide pharmacology
- Abstract
Although estrogen can induce LH hypersecretion in ovariectomized (ovx) rats, the magnitude and duration of the LH surge are invariably far less than those of the preovulatory LH surge in cycling rats. Recently, the crucial roles of hypothalamic nitric oxide (NO) and neuropeptide Y (NPY) in the induction of LHRH and the LH surge have been recognized. In this study, we have tested the hypothesis that low grade stimulatory feedback effects of 17 beta-estradiol (E2) on LH secretion in ovx rats is due to a deficit in NO signaling. As the NO substrate, L-arginine (L-Arg), can enhance NO-dependent physiological responses, the effects of L-Arg on the LH surge in E2-primed ovx rats was studied. Ovx rats bearing permanent cannulas in the lateral cerebroventricle received 15-mm long SILASTIC capsules filled with E2 (300 micrograms/ml oil, s.c.) at 1000 h. Two days later, saline (SAL) or L-Arg (0.1 or 1 nmol in 5 microliters SAL) or D-Arg (1.0 nmol) was injected intracerebroventricularly every hour from 1100-1400. Blood samples were collected at 1100 h and at hourly intervals from 1400-1800 h via intraatrial cannulae implanted the day before. Whereas an expected moderate LH surge occurred in SAL-injected control rats, L-Arg, but not D-Arg, greatly augmented the magnitude and duration of the LH surge, with peak values attaining the range normally seen on proestrus. Furthermore, L-Arg (1 or 10.0 mM) and the NO donor, sodium nitroprusside (1 or 10.0 mM), stimulated the basal and K(+)-induced in vitro release of LHRH and NPY from the hypothalami of ovarian steroid-primed ovx rats. As NPY is stimulatory to LHRH release, we next tested the possibility that L-Arg/NO may initially stimulate NPY release in the hypothalamus, which, in turn, elicits LHRH discharge. E2-treated ovx rats receiving intracerebroventricular L-Arg were injected with antisense or missense oligodeoxynucleotides (oligos) to NPY-messenger RNA. Antisense, not missense, NPY oligos blocked the L-Arg-induced potentiation of the LH surge. These results revealed for the first time the neurochemical cause underlying the E2 feedback action and show that a deficiency in either L-Arg itself or L-Arg-based NO signaling is responsible for the low grade LH surge in ovx rats treated with estrogen alone. Additionally, the L-Arg-dependent potentiation of the LH surge may involve increased signaling in the NO and NPYergic systems, resulting in optimal LHRH hypersecretion from the hypothalamus.
- Published
- 1996
- Full Text
- View/download PDF
30. Disinhibition from opioid influence augments hypothalamic neuropeptide Y (NPY) gene expression and pituitary luteinizing hormone release: effects of NPY messenger ribonucleic acid antisense oligodeoxynucleotides.
- Author
-
Xu B, Sahu A, Kalra PS, Crowley WR, and Kalra SP
- Subjects
- Animals, Base Sequence, Female, Hypothalamus, Middle metabolism, Injections, Intraventricular, Molecular Sequence Data, Oligonucleotide Probes genetics, RNA, Messenger metabolism, Rats, Arcuate Nucleus of Hypothalamus metabolism, Luteinizing Hormone blood, Median Eminence metabolism, Naloxone pharmacology, Neuropeptide Y genetics, Neuropeptide Y metabolism, Oligonucleotides, Antisense pharmacology, RNA, Messenger genetics
- Abstract
It is well known that hypothalamic endogenous opioid peptides (EOP) exert an inhibitory influence on LH release, and that a restraint on this inhibitory tone triggers preovulatory LHRH and LH hypersecretion. Recent evidence suggests that hypothalamic neuropeptide Y (NPY) is an important component of the neural circuitry that participates in induction of the LH surge. We have reported previously that blockade of EOP influence by the opioid receptor antagonist, naloxone (NAL), stimulated NPY accumulation in the median eminence (ME) in association with increased LH release in estradiol-17 beta-primed ovariectomized rats. To evaluate whether a restraint on the EOP system will result in an increase in NPY synthesis, the effects of NAL infusion on preproNPY messenger RNA (mRNA) levels in the medial basal hypothalamus were studied by solution hybridization/RNase protection assay and by in situ hybridization. NAL (2 mg/0.6 ml.h) or saline (SAL, 0.6 ml/h) was infused for 3 h (1100-1400 h) via an intrajugular cannula in estradiol-17 beta-primed ovariectomized rats. In accord with previous studies, NAL infusion significantly increased plasma LH levels at 1400 h. concomitant with this activation of LH release, NPY gene expression was also augmented. As compared with initial control levels at 1100 h, preproNPY mRNA levels in the medial basal hypothalamus increased at 1400 h in SAL (106%)- and NAL (202%)-infused rats, and at this time, preproNPY mRNA levels were significantly higher in NAL-infused rats than in SAL-infused rats. In situ hybridization studies showed that NAL infusion significantly increased the preproNPY mRNA signal at 1400 h mainly in the rostral and middle regions of the arcuate nucleus (ARC) as compared with that seen at 1100 and 1400 h in SAL-infused rats. To examine further the relationship between the NAL-induced increase in LH release and increase in NPY gene expression, the effects of NPY mRNA antisense oligonucleotides (oligos) on NPY levels in the ME-ARC and on plasma LH levels were studied in NAL-infused rats. In NAL-infused rats, intracerebroventricular administration of a NPY antisense oligo (25 micrograms/rat) at 1000, 1200, and 1300 h decreased NPY levels in the ME-ARC and blocked the increase in plasma LH levels at 1500 h, whereas control missense oligos had no effect. Collectively, these results show that a decrease in opioid influence rapidly augments NPY gene expression in a subpopulation of neurons in the ARC, and support the hypothesis that a disinhibition from opioid influence acutely promotes NPY synthesis and release, which is necessary for phasic LH discharge in rats.
- Published
- 1996
- Full Text
- View/download PDF
31. Insulin and insulin-like growth factor II suppress neuropeptide Y release from the nerve terminals in the paraventricular nucleus: a putative hypothalamic site for energy homeostasis.
- Author
-
Sahu A, Dube MG, Phelps CP, Sninsky CA, Kalra PS, and Kalra SP
- Subjects
- Animals, Feeding Behavior, Homeostasis, In Vitro Techniques, Insulin-Like Growth Factor I pharmacology, Male, Paraventricular Hypothalamic Nucleus metabolism, Rats, Rats, Sprague-Dawley, Energy Metabolism, Insulin pharmacology, Insulin-Like Growth Factor II pharmacology, Neuropeptide Y metabolism, Paraventricular Hypothalamic Nucleus drug effects
- Abstract
It has been recently recognized that a distinct signaling pathway in the hypothalamus is involved in the stimulation of feeding in mammals. Neuropeptide Y (NPY), a member of the pancreatic polypeptide family, is the most potent orexigenic signal, and its secretion in discrete hypothalamic sites increases in response to insulinopenia produced by food deprivation or experimental diabetes. To establish the site of interaction between the hypothalamus and the pancreas, we examined the effects of insulin on NPY release in vivo and in vitro from hypothalamic sites known to be involved in feeding behavior. In the first study we evaluated the effects of peripheral insulin injections (1 U/kg.day, sc) on NPY levels in seven hypothalamic nuclei in food-deprived (FD) and ad libitum-fed rats. Whereas food deprivation for 3 days increased NPY levels in the medial preoptic area, paraventricular nucleus (PVN), and arcuate nucleus, insulin injections, which did not alter blood glucose levels, returned NPY levels to the control range selectively in the PVN. NPY levels in the hypothalamic nuclei remained unchanged after insulin injections in ad libitum-fed rats. The in vivo NPY release in the PVN of FD rats, evaluated by the push-pull cannula technique, also decreased in response to peripheral insulin injections. Finally, the effects of insulin, insulin-like growth factor I (IGF-I), and IGF-II on NPY release in vitro from the microdissected PVN and two central neighboring sites, the ventromedial nucleus and the median eminence-arcuate nucleus, of FD rats were evaluated. Both insulin (0.67 or 6.7 nM) and IGF-II (0.7 or 7.0 nM) decreased the release of NPY in a dose-dependent manner only from the PVN. On the other hand, IGF-I (0.07 or 7.0 nM) failed to alter the basal PVN NPY efflux. As the PVN is richly innervated by NPY-containing nerve terminals, the results of these in vivo and in vitro studies suggest that the site of insulin action on the hypothalamic NPY network may reside at the level of PVN nerve terminals or at the interneurons in contact with NPY nerve terminals. Although insulin may have a direct effect in reducing NPY release from the PVN, the effectiveness of IGF-II in decreasing NPY release from the PVN raises the possibility that insulin's action may also be mediated via hypothalamic IGF-II neuronal pathways.
- Published
- 1995
- Full Text
- View/download PDF
32. Anorectic effects of estrogen may be mediated by decreased neuropeptide-Y release in the hypothalamic paraventricular nucleus.
- Author
-
Bonavera JJ, Dube MG, Kalra PS, and Kalra SP
- Subjects
- Animals, Eating drug effects, Estradiol metabolism, Female, Hypothalamus drug effects, Hypothalamus metabolism, Paraventricular Hypothalamic Nucleus drug effects, Potassium Chloride pharmacology, Rats, Weight Gain drug effects, beta-Endorphin metabolism, Appetite Depressants, Estradiol pharmacology, Neuropeptide Y metabolism, Paraventricular Hypothalamic Nucleus physiology
- Abstract
There is a considerable body of evidence to suggest that estrogen suppresses food intake and body weight gain by an action in the hypothalamus. However, the neurotransmitter/neuromodulator mediating the anorectic effects of estrogen are unknown. Neuropeptide-Y (NPY) is the most potent orexigenic signal known, and NPY-producing neurons in the hypothalamus concentrate 17 beta-estradiol (E2). In these studies we tested the hypothesis that estrogen-induced anorectic effects may be due to decreased NPY levels and release in hypothalamic sites previously implicated in the control of food intake. The results show that uninterrupted physiological levels of E2 in ovariectomized rats suppressed daily food intake and body weight gain. Evaluation of NPY concentrations in five hypothalamic sites showed that NPY levels were decreased selectively in the paraventricular nucleus (PVN) and neighboring perifornical nucleus of E2-treated rats. In contrast, concentrations of beta-endorphin, another less potent orexigenic peptide, were not changed by E2 in any hypothalamic site. In the next experiment, the effects of similar E2 treatment on NPY release in vitro from the PVN and ventromedial nucleus were studied in rats killed at the onset of the dark phase when food intake increases in conjunction with increased PVN NPY secretion. The results show that basal and KCl-induced NPY release were significantly decreased from the PVN of E2-treated compared to those in control rats. In contrast, both basal and KCl-induced NPY release from the ventromedial nucleus of E2-treated rats were similar to those in control rats. Collectively, these results show that estrogen suppresses NPY levels and release selectively from the PVN. As NPY levels and release in the PVN have been shown to be highly correlated with appetite status, and the PVN is one of the important sites of NPY action, these findings imply that the anorectic effects of estrogen may be mediated by decreased NPY release from the PVN NPY innervations.
- Published
- 1994
- Full Text
- View/download PDF
33. Hypothalamic neuropeptide-Y gene expression increases before the onset of the ovarian steroid-induced luteinizing hormone surge.
- Author
-
Sahu A, Crowley WR, and Kalra SP
- Subjects
- Animals, Female, Gonadotropin-Releasing Hormone genetics, Ovariectomy, RNA, Messenger analysis, Rats, Rats, Sprague-Dawley, Gene Expression Regulation, Gonadal Steroid Hormones pharmacology, Hypothalamus, Middle metabolism, Luteinizing Hormone blood, Neuropeptide Y genetics
- Abstract
Neuropeptide-Y (NPY), a hypothalamic peptide, is involved in stimulation of LHRH and LH surges during proestrus and those induced by ovarian steroids in ovariectomized (ovx) rats. The NPY neurons that reside in the arcuate nucleus of the medial basal hypothalamus (MBH) and accumulate 17 beta-estradiol participate in the initiation of LHRH and LH surges. To determine whether NPY synthesis is altered in conjunction with the LH surge, we studied the dynamic changes in prepro-NPY mRNA levels in the MBH in association with the LH surge elicited by estradiol benzoate (EB) alone or by progesterone (P) in EB-primed ovx rats. Five days after ovariectomy, rats received oil or EB (30 micrograms/rat) at 1000 h on day 0. On day 2, these rats were injected with either oil or P (2 mg/rat) at 1000 h. Rats were killed before (1000 h) and at 2-h intervals after oil or P injection. The MBHs were dissected out and processed for determination of prepro-NPY mRNA levels by solution hybridization/RNase protection assay using a cRNA probe. Although in control ovx rats, prepro-NPY mRNA levels remained unchanged between 1000-1600 h, prepro-NPY mRNA levels showed dynamic changes in steroid-primed rats. In the EB-primed rats, prepro-NPY mRNA levels rose significantly (100%) at 1200 and 1400 h before the LH rise at 1600 h, and the levels remained elevated up to 1800 h. After P injection to the EB-primed rats, this response was further augmented, with a slightly different temporal pattern. Prepro-NPY mRNA levels rose at 1400 h (600%) before the onset of the LH rise at 1600 h and declined steadily to significantly lower values at 1800 h, coincident with the highest rate of LH secretion. These studies demonstrate dynamic shifts in hypothalamic NPY gene expression in association with the LH (LHRH) surge, and that maximal increases occur before the onset of the LH rise, but thereafter, NPY gene expression diverged in the two ovarian-steroid treatment models. These findings along with previous evidence of similar antecedent increases in NPY content in the median eminence, followed by release, suggest that augmented NPY synthesis and release are two temporally dissociable neural events for the LHRH and LH surges.
- Published
- 1994
- Full Text
- View/download PDF
34. Role of galanin in stimulation of pituitary luteinizing hormone secretion as revealed by a specific receptor antagonist, galantide.
- Author
-
Sahu A, Xu B, and Kalra SP
- Subjects
- Animals, Arcuate Nucleus of Hypothalamus drug effects, Cerebral Ventricles drug effects, Circadian Rhythm drug effects, Estradiol administration & dosage, Female, Galanin, In Vitro Techniques, Injections, Intraventricular, Median Eminence drug effects, Neuropeptides pharmacology, Ovariectomy, Peptides administration & dosage, Pituitary Gland drug effects, Progesterone administration & dosage, Rats, Rats, Inbred Strains, Receptors, Galanin, Arcuate Nucleus of Hypothalamus metabolism, Cerebral Ventricles physiology, Estradiol pharmacology, Gonadotropin-Releasing Hormone metabolism, Luteinizing Hormone metabolism, Median Eminence metabolism, Peptides pharmacology, Pituitary Gland metabolism, Progesterone pharmacology, Receptors, Gastrointestinal Hormone antagonists & inhibitors, Substance P analogs & derivatives
- Abstract
In this study, a specific galanin (GAL) receptor antagonist, galantide, was employed to evaluate the role of endogenous GAL in episodic basal and phasic LH release in rats. To assess the specificity of galantide, a series of experiments was performed. In the first experiment, we observed that administration of GAL (0.62 nm) intracerebroventricularly (icv) in ovarian steroid-primed ovariectomized (ovx) rats rapidly increased plasma LH levels between 10-30 min, and prior injection of galantide icv (5 nm) blocked the GAL-induced LH release. In the second experiment, galantide inhibited the GAL-evoked in vitro release of LHRH from the median eminence-arcuate nucleus (ME-ARC) of ovarian steroid-primed ovx rats. In addition, galantide on its own significantly decreased the basal efflux of LHRH from the ME-ARC of similarly treated rats, thereby suggesting that even the basal LHRH secretion may be a GAL-dependent event. In the third experiment, the effects of galantide on the phasic LH surge elicited by progesterone (P) in estradiol benzoate-primed ovx rats and that occurring spontaneously on proestrus were examined. Ovx rats bearing icv cannulae were primed with estradiol benzoate (30 micrograms/rat, sc) and 2 days later received a P (2 mg/rat, sc) injection at 1000 h to evoke a LH surge in the afternoon. Galantide (1 or 5 nm) in 3 microliters saline or saline was injected icv at 1300, 1400, and 1500 h. The results showed that the two dosages of galantide suppressed the LH surge in the afternoon. On the other hand, only a very high dose of galantide (15 nm) injected iv at 1300, 1400, and 1500 h blunted the P-induced LH hypersecretion. Central injections of galantide (1 or 5 nm) at 1300, 1400, and 1500 h on proestrus also inhibited the preovulatory LH surge and significantly reduced the numbers of rats ovulating the following day. In the final experiment, the role of GAL receptors in modulation of episodic LH release was analyzed in ovx rats. The results showed that an injection of galantide (5 nm, icv) significantly decreased both the mean LH levels and the amplitude of LH episodes during the 3-h observation period. Cumulatively, these results show that normally GAL stimulates LHRH release by activation of a specific receptor located in the ME-ARC, and that GAL may be a key excitatory signal in the hypothalamic neural circuitry involved in the regulation of basal episodic and phasic LHRH secretion in cycling female rats.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1994
- Full Text
- View/download PDF
35. Evidence that nitric oxide may mediate the ovarian steroid-induced luteinizing hormone surge: involvement of excitatory amino acids.
- Author
-
Bonavera JJ, Sahu A, Kalra PS, and Kalra SP
- Subjects
- Animals, Arcuate Nucleus of Hypothalamus metabolism, Arginine analogs & derivatives, Arginine pharmacology, Estradiol pharmacology, Female, Hypothalamus, Middle metabolism, N-Methylaspartate pharmacology, NG-Nitroarginine Methyl Ester, Nitric Oxide antagonists & inhibitors, Nitroprusside pharmacology, Ovariectomy, Pituitary Gland metabolism, Preoptic Area metabolism, Rats, Luteinizing Hormone metabolism, N-Methylaspartate physiology, Nitric Oxide physiology, Progesterone pharmacology
- Abstract
The involvement of excitatory N-methyl-D-aspartate (NMDA) receptors in the hypothalamic control of pituitary LH secretion is well recognized. Recent evidence shows that nitric oxide (NO), a free radical gas, may act as neurotransmitter in the brain, and its efflux is stimulated by activation of NMDA receptors. Studies were undertaken to determine whether NO is involved in the hypothalamic release of LHRH and in the LH surge induced by progesterone (P) in estrogen-primed ovariectomized rats. Rats were ovariectomized and 2 weeks later received estradiol benzoate (30 micrograms sc) at 1000 h. Two days later, P was injected at 1000 h to potentiate the estradiol benzoate-induced LH surge in the afternoon. Serial blood samples were collected at hourly intervals from 1400-1800 h via an intraatrial cannula implanted the day before P injection. Additionally, at various times before onset of the LH surge at 1400 h, the rats were injected sc with one of three inhibitors of NO synthase, the enzyme that generates NO. Control, saline-injected rats showed unambiguous LH surges in the afternoon. However, either a single injection at 1000 h of NG-methyl-L-arginine (20 mg/kg) or three injections at 1000, 1200, and 1400 h of either Nw-nitro-L-arginine methyl ester (NAME, 40 mg/kg) or Nw-nitro-L-arginine (60 mg/kg) to inhibit NO efflux markedly suppressed the P-induced LH surge in the afternoon. To ascertain whether suppression of LH surge was due to blockade of hypothalamic LHRH release, a series of in vitro studies were performed in steroid-primed rats. First we examined the effects of sodium nitroprusside (NPS), a compound that spontaneously generates and releases NO. NPS increased basal and KCl-induced LHRH release in vitro from the medial basal hypothalamus-preoptic area and median eminence fragments. No direct effect of NO at the pituitary level was seen, since NPS did not alter basal or LHRH-induced LH in vitro release from hemipituitaries. In addition, we tested the effects of NAME on NMDA-induced LHRH release in vitro from the median eminence-arcuate nucleus fragments. As expected, NMDA alone (50 mM) induced a significant increase in LHRH release. Addition of NO synthase inhibitor, NAME (1 or 10 mM) to suppress NO efflux, significantly diminished the NMDA-induced LHRH release.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1993
- Full Text
- View/download PDF
36. Mandatory neuropeptide-steroid signaling for the preovulatory luteinizing hormone-releasing hormone discharge.
- Author
-
Kalra SP
- Subjects
- Animals, Feedback, Female, Humans, Hypothalamus physiology, Male, Estrogens physiology, Gonadotropin-Releasing Hormone metabolism, Neuropeptides physiology, Proestrus physiology, Progesterone physiology, Signal Transduction physiology
- Abstract
In this article I have examined various aspects of the complex spatio-temporal patterning of peptidergic signaling that lead to synchronized development of neural events for the preovulatory LHRH discharge on proestrus. Undoubtedly, the integration of these events is orchestrated by both ovarian steroids, E2 and P. Evidence accumulated in recent years has failed to affirm the perceived notion that E2 is an adequate peripheral signal for the timely, robust discharge of LHRH on proestrus. The current understanding is shaped by the thesis that the concerted central actions of E2 and P are mediated by a host of regulatory peptides produced locally in the hypothalamus, and steroids, in general, augment the production and release of both inhibitory and excitatory peptides in a timely fashion to facilitate the preovulatory LHRH discharge. Since these peptidergic pathways appear mandatory for signal transfer, considerable recent research has been devoted first to identifying the signals that selectively participate in the induction of preovulatory LHRH (LH) surge, and then to trace the route of signal transmission that ultimately leads to LHRH hypersecretion on the afternoon of proestrus (Fig. 1). The peptidergic pathways that propagate and transmit impulses for the preovulatory LHRH discharge reside in the SCN-MPN-MPOA-ARC-ME neural complex (Fig. 1). The timely initiation of these impulses is entrained to the photo-periodic input reaching the SCN by the retino-hypothalamic tract. The evidence is already in place to show that further information processing is transduced in the MPN; however, the nature of neurochemical signaling between the two sites remains to be deciphered. The available evidence favors a mandatory participation of inhibitory (EOP and NPK) and excitatory (NPY, GAL, NT, and AII) messenger molecules within the SCN-MPN-MPOA-ARC-ME complex (Fig. 1). It is possible that the relevant information from the SCN-MPN is conveyed caudally to the ARC in order to initiate a chain of events for disinhibition/excitation of the NPY-EOP network and to affect LHRH neurosecretion at the perikaryal level in the MPOA and at axon terminals in the ME. Also, either concurrently or on a time-delayed basis, the relevant information from the MPN may be relayed to the MPOA via the local peptidergic network comprised of NT, EOP, NPK, and GAL. This transmission may initially be critical for elicitation of antecedent neurosecretory events in the ME and to ultimately evoke the preovulatory LHRH surge.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1993
- Full Text
- View/download PDF
37. Role of neuropeptide-Y in episodic luteinizing hormone release in ovariectomized rats: an excitatory component and opioid involvement.
- Author
-
Xu B, Sahu A, Crowley WR, Leranth C, Horvath T, and Kalra SP
- Subjects
- Animals, Corticotropin-Releasing Hormone pharmacology, Female, Naloxone pharmacology, Periodicity, Rats, Luteinizing Hormone metabolism, Neuropeptide Y pharmacology, Ovariectomy, beta-Endorphin physiology
- Abstract
We tested the hypothesis that hypothalamic neuropeptide-Y (NPY) is an excitatory signal in the episodic secretion of LH in ovariectomized (ovx) rats and that the suppression of LH secretion that consistently follows intracerebroventricular administration of NPY is due to concurrent release of opioids or CRH, both previously shown to readily inhibit LH release. In the first experiment, ovx rats received continuous intraventricular infusion of either serum containing NPY antibodies (NPY-Ab) or normal rabbit serum (control) at dilutions of 1:5 or 1:1. NPY-Ab infusion at a 1:5 dilution significantly decreased mean plasma LH levels and LH pulse amplitude without affecting LH pulse frequency over a 3-h period of observation. However, infusion of relatively more concentrated NPY-Ab (1:1) markedly decreased not only mean plasma LH levels and LH pulse amplitude, but also the frequency of LH episodes. In the next experiment, we observed that intraventricular administration of NPY (0.2 nmol) suppressed LH release for 60 min. However, blockade of opiate receptors with iv infusion of naloxone (2 mg/h) before and after NPY injection completely counteracted the NPY-induced inhibition of LH release. On the other hand, prior blockade of the CRH receptors with alpha-helical CRH-(9-41) (25 or 100 micrograms/rat) was ineffective in reversing the inhibitory LH response of NPY (0.125 nmol). These results together with our previous demonstration of morphological communication between NPY and beta-endorphin neurons, show that suppression of LH by exogenous NPY in ovx rats may result from concurrent stimulation of opioids, primarily beta-endorphin. However, diminution of all parameters of episodic LH secretion by NPY-Ab affirms the notion that the NPY network is a physiologically important excitatory component of the hypothalamic pulse generator circuitry that regulates episodic LH secretion in rats.
- Published
- 1993
- Full Text
- View/download PDF
38. Evidence that luteinizing hormone suppression in response to inhibitory neuropeptides, beta-endorphin, interleukin-1 beta, and neuropeptide-K, may involve excitatory amino acids.
- Author
-
Bonavera JJ, Kalra SP, and Kalra PS
- Subjects
- Animals, Injections, Intraventricular, Interleukin-1 administration & dosage, Kinetics, Male, N-Methylaspartate administration & dosage, Neuropeptides administration & dosage, Rats, beta-Endorphin administration & dosage, Interleukin-1 pharmacology, Luteinizing Hormone metabolism, N-Methylaspartate pharmacology, Neuropeptides pharmacology, Tachykinins, beta-Endorphin pharmacology
- Abstract
A large body of recent evidence suggests that a number of inhibitory and excitatory neuropeptides and amino acids may participate in the episodic secretion of hypothalamic LHRH and pituitary LH in castrated rats. However, the precise functional relationships among these messenger molecules in the control of LH secretion remain to be ascertained. The aim of this study was to test the hypothesis that inhibition of LH release by an opioid [beta-endorphin (beta END)], cytokine [interleukin-1 beta (IL-1 beta)], or tachykinin [neuropeptide-K (NPK)] is a result of diminished excitatory amino acid (EAA) signaling. Adult male rats were castrated and received an intracerebroventricular cannula in the third ventricle for administration of beta END (10 micrograms/rat), NPK (2.5 nmol/rat), or IL-1 beta (100 ng/rat) 2 weeks postcastration. One day before the experiments, rats received an intraatrial cannula for frequent blood sampling and for iv injection of the glutamate receptor agonist N-methyl-D-aspartate (NMDA; 5 mg/kg) at 30-min intervals. Blood samples for LH measurements were withdrawn immediately before and 10 min after each NMDA injection. The results show that intracerebroventricular beta END, IL-1 beta, or NPK inhibited LH release. Multiple injections of NMDA did not alter the existing pattern of LH secretion in castrated control rats. However, similar NMDA injections completely prevented the decrease in LH release by beta END, IL-1 beta, or NPK. Plasma LH levels in these rats remained within the range seen in untreated control rats throughout the 120-min duration of the experiment, and NMDA injections at 30-min intervals evoked pulses of LH that resembled those seen normally in castrated rats. The blockade of the inhibitory effects of the three peptides by NMDA and previous knowledge of hypothalamic sites of NMDA action suggest that EAA systems may represent a common pathway down-stream in the hypothalamic LHRH-regulating circuitry to mediate diminution of LH release by inhibitory peptides. Further, their inhibitory influence may be exerted either directly at the level of LHRH neurons and/or by diminution in EAA efflux, leading to suppression of LHRH and LH release.
- Published
- 1993
- Full Text
- View/download PDF
39. Dynamic changes in hypothalamic angiotensin II levels and release in association with progesterone-induced luteinizing hormone surge.
- Author
-
Phillips MI, Wang H, Kimura B, Rejtman M, Koduri P, and Kalra SP
- Subjects
- Angiotensin II cerebrospinal fluid, Animals, Estrogens pharmacology, Female, Ovariectomy, Paraventricular Hypothalamic Nucleus metabolism, Perfusion, Rats, Rats, Sprague-Dawley, Angiotensin II metabolism, Hypothalamus metabolism, Luteinizing Hormone metabolism, Progesterone pharmacology
- Abstract
To test the hypothesis that brain angiotensin II (Ang II) may be involved in the preovulatory release of LH on proestrus, we evaluated the pattern of changes in hypothalamic Ang II levels and release in ovariectomized (ovx) rats treated sequentially with estrogen and progesterone. This is an experimental paradigm that reliably evokes dynamic changes in hypothalamic LHRH levels in association with LH hypersecretion, simulating the LH surge on proestrus. Rats were ovx and after 4 weeks received estradiol benzoate followed by progesterone 2 days later at 1000 h. We observed that in these progesterone-treated rats, serum LH levels were low until 1400 h, but thereafter, the rate of LH secretion increased and remained elevated at 1600 h when the experiment was terminated. In these rats, hypothalamic Ang II levels increased abruptly at 1330 h and returned rapidly to baseline levels before the onset of LH surge. Also, a similar pattern in hypothalamic Ang II levels occurred at 1500 h with the rise and peak serum LH levels in the late afternoon. In the second experiment, Ang II levels in the cerebrospinal fluid (CSF) of rats similarly pretreated with ovarian steroids were evaluated. Again, CSF Ang II levels rose abruptly to a peak at 1330 h and returned to baseline range preceding the expected rise in serum LH. Thereafter, no further change in CSF Ang II levels was detected during the period of LH hypersecretion. In the third experiment, perfusates were collected from a push-pull cannula aimed at the paraventricular nucleus in ovx rats similarly treated with ovarian steroids. A peak of Ang II was observed at 1330 h and a later peak at 1430 h. A comparison with LH profiles indicated that these peaks in Ang II levels were evident before and during the LH surge. Thus, in three separate experiments, the results showed that rapid dynamic changes in hypothalamic Ang II levels and release occur in association with the progesterone-induced LH surge in estrogen-primed ovx rats. These findings support the previous observations that Ang II can stimulate LHRH and LH release. Since similar, temporally correlated changes occur in hypothalamic neuropeptide Y and LHRH, the peptides involved in the induction of LH surge, these results are in agreement with the hypothesis that Ang II-expressing neurons may play an important role in the hypothalamic circuitry responsible for stimulation of LH surge in ovarian steroid-treated ovx rats.
- Published
- 1993
- Full Text
- View/download PDF
40. Neuropeptide Y release from the paraventricular nucleus increases in association with hyperphagia in streptozotocin-induced diabetic rats.
- Author
-
Sahu A, Sninsky CA, Phelps CP, Dube MG, Kalra PS, and Kalra SP
- Subjects
- Animals, Diabetes Mellitus, Experimental physiopathology, Male, Paraventricular Hypothalamic Nucleus drug effects, Potassium Chloride pharmacology, Rats, Rats, Sprague-Dawley, Diabetes Mellitus, Experimental complications, Hyperphagia etiology, Neuropeptide Y metabolism, Paraventricular Hypothalamic Nucleus metabolism
- Abstract
We tested the hypothesis that the hyperphagia observed in streptozotocin (STZ)-induced diabetic rats is due to increased release of neuropeptide Y (NPY) in the paraventricular nucleus (PVN) of the hypothalamus. In the first experiment, male rats were injected with STZ or vehicle (control) via the tail vein and 18-20 days later, NPY levels in seven hypothalamic sites and release in vitro from selected hypothalamic sites were evaluated. The results showed that in association with STZ-produced marked hyperglycemia and hyperphagia, NPY concentrations were increased in four hypothalamic sites, including the PVN. Evaluation of NPY release in vitro showed that both basal and KCl-induced release was significantly higher from the micro-dissected PVN of STZ-treated than control rats. A similar augmentation in the NPY efflux in vitro was detected from the median eminence arcuate nucleus, but not from the neighboring ventromedial nucleus of STZ-treated rats. In the second experiment, rats were treated with STZ or vehicle and received permanent push-pull cannula (PPC) in the PVN for evaluation of NPY release in vivo 18-21 days after STZ treatment. The results showed that mean NPY levels in the perfusates collected from the PVN of diabetic rats were significantly higher as compared to control rats. Since NPY is the most potent naturally occurring orexigenic signal and the PVN is an important initial site of NPY action in the stimulatory pathway regulating feeding, our findings of augmented PVN NPY release in vivo and in vitro are in accord with the hypothesis that increased NPY secretion in the PVN may be responsible for hyperphagia in diabetic rats.
- Published
- 1992
- Full Text
- View/download PDF
41. Neuropeptide-Y innervation of beta-endorphin-containing cells in the rat mediobasal hypothalamus: a light and electron microscopic double immunostaining analysis.
- Author
-
Horvath TL, Naftolin F, Kalra SP, and Leranth C
- Subjects
- 3,3'-Diaminobenzidine, Animals, Colchicine pharmacology, Female, Gonadotropin-Releasing Hormone analysis, Hypothalamus, Middle ultrastructure, Immunohistochemistry, Luteinizing Hormone metabolism, Microscopy, Electron methods, Ovariectomy, Rats, Rats, Sprague-Dawley, Hypothalamus, Middle chemistry, Hypothalamus, Middle cytology, Neuropeptide Y analysis, beta-Endorphin analysis
- Abstract
Central administration of neuropeptide-Y (NPY) inhibits pituitary LH release in ovariectomized rats and stimulates LH release in intact and ovariectomized rats pretreated with ovarian steroids. Although the precise neural mechanism of this dual effect of NPY is not known, experimental evidence suggests an underlying interaction between hypothalamic NPY and the inhibitory beta-endorphin (beta END) systems in the neuroendocrine regulation of pituitary LH release in the rat. The present study was undertaken to examine the morphological basis of the interaction between these two peptidergic systems in the hypothalamus. Sections of the mediobasal hypothalamus of colchicine-pretreated female rats were double immunostained for NPY and beta END and examined by light and electron microscopy. The light brown diaminobenzidine reaction was used to visualize beta END cells, while NPY neurons were labeled with a dark blue nickel ammonium sulfate-intensified diaminobenzidine reaction. Under the light microscope, a dense network of NPY-immunoreactive axons and axon terminals was observed in close apposition with beta END-immunoreactive neurons throughout the medial basal hypothalamus. Electron microscopic examination revealed that NPY-immunoreactive boutons formed axosomatic and axo-dendritic synaptic connections with beta END cells. A majority of these synaptic membrane specializations appeared asymmetrical [corrected]. In light of the previous evidence of excitatory and inhibitory effects on LH release and the existence of direct synaptic connections between NPY and LHRH neurons in the hypothalamus, the current results imply that the dual effects of NPY on LH secretion may involve modulation of LHRH secretion, both by the direct route and indirectly through the hypothalamic beta END system.
- Published
- 1992
- Full Text
- View/download PDF
42. Diverse effects of tachykinins on luteinizing hormone release in male rats: mechanism of action.
- Author
-
Kalra PS, Sahu A, Bonavera JJ, and Kalra SP
- Subjects
- Animals, Cerebral Ventricles drug effects, Dose-Response Relationship, Drug, Injections, Intraventricular, Luteinizing Hormone blood, Male, Neurokinin A administration & dosage, Neurokinin B administration & dosage, Orchiectomy, Peptide Fragments administration & dosage, Rats, Rats, Inbred Strains, Reference Values, Substance P administration & dosage, Tachykinins administration & dosage, Cerebral Ventricles physiology, Luteinizing Hormone metabolism, Neurokinin A pharmacology, Neurokinin B pharmacology, Neuropeptides pharmacology, Peptide Fragments pharmacology, Substance P pharmacology, Tachykinins pharmacology
- Abstract
The tachykinins are a group of structurally related peptides found in the rat hypothalamus and anterior pituitary. We have evaluated the effects of four tachykinins on LH release in male rats. In intact male rats, intracerebroventricular (icv) injection of neurokinin A (NKA), neuropeptide K (NPK), and neuropeptide-gamma (NP gamma) elicited dose-related, transient increases in plasma LH. Substance P (SP) was ineffective under these conditions. A further examination showed that in vitro incubation with either NPK or NP gamma of hemipituitaries from intact but not castrated male rats promoted release of LH into the medium, thereby revealing that the excitatory effects of tachykinins in intact male rats may, in part, be a result of stimulation of LH release directly from the anterior pituitary. On the other hand, the effects of these four tachykinins on LH release were different in castrated rats. Intracerebroventricular injection of NPK, NKA, and NP gamma as well as SP, which was ineffective in intact male rats, evoked a long-lasting suppression of LH release. Comparatively, NPK was the most effective tachykinin in eliciting LH responses in both of these tests involving different endocrine environments. We next evaluated the possibility that the inhibitory effects of tachykinins (NPK) may be mediated by activation of inhibitory endogenous opioid peptides. The results showed that iv infusion of the opiate receptor antagonist naloxone, to block the possible inhibitory effects of endogenous opioid peptides, only partially counteracted the suppressive effects of icv NPK on plasma LH levels. Thus, in addition to revealing the diverse effects of structurally related tachykinins on LH release, the results of these investigations showed specifically that the NK-2 receptor agonists NPK, NP gamma, and NKA stimulated LH release in intact rats, in part, by a direct action at the level of the pituitary, whereas the NK-1 receptor agonist SP was inactive under these conditions. These findings imply a paracrine/autocrine mode of excitatory action on LH release involving pituitary NK-2 receptor subtypes. On the other hand, in castrated rats, all four tachykinins readily suppressed LH release by a central action involving, in part, an activation of hypothalamic opioid systems.
- Published
- 1992
- Full Text
- View/download PDF
43. Neuropeptide Y release is elevated from the microdissected paraventricular nucleus of food-deprived rats: an in vitro study.
- Author
-
Dube MG, Sahu A, Kalra PS, and Kalra SP
- Subjects
- Animals, In Vitro Techniques, Male, Paraventricular Hypothalamic Nucleus drug effects, Potassium Chloride pharmacology, Rats, Rats, Inbred Strains, Food Deprivation, Neuropeptide Y metabolism, Paraventricular Hypothalamic Nucleus metabolism
- Abstract
Intracerebroventricular injection of neuropeptide Y (NPY) stimulates a robust dose-related feeding response in the rat. Experimental evidence attests to the view that the release of NPY in the paraventricular nucleus (PVN), a site richly innervated by NPY immunopositive fibers, is responsible for stimulation of feeding behavior. However, there is little information on the neuroendocrine factors involved in regulation of NPY release, in part due to the unavailability of reliable techniques to monitor PVN NPY release. In this study, we have validated an in vitro technique to assess NPY release from the PVN and other neighboring hypothalamic sites of the rat brain. In the first experiment, freshly dissected brains from male rats were processed for 300-microns thick sections with a vibratome. The PVNs were microdissected from the brain sections under a stereomicroscope and incubated in 250 microliters Krebs Ringer bicarbonate buffer at 37 C for basal and KCl-induced NPY release. The results showed that basal NPY efflux from the excised PVN was detectable and increased in relation to the number of PVNs in the incubation chambers. Addition of KCl at the end of the 60-min basal incubation period increased NPY release further, the increments were again closely related to the number of PVN punches in the incubation chambers. In the second experiment, the assumption that in vitro basal and KCl-evoked NPY release from the PVN reflected the in vivo pattern of PVN NPY secretion was validated. The effects of 4-day food deprivation (FD), an experimental paradigm known to augment in vivo PVN NPY secretion, on the in vitro NPY release from PVN and ventromedial nucleus were evaluated. The results showed that both basal and KCl-evoked NPY release was significantly higher from the PVN of food-deprived than control rats on ad libitum rat chow. This FD-induced incremental NPY response was site-specific because the basal and KCl-evoked NPY effluxes from the ventromedial nucleus of FD and control rats were similar. Thus, in agreement with previous in vivo findings, NPY release in vitro is also augmented selectively from the PVN in response to fasting. Cumulatively, these results demonstrate that NPY release in vitro from hypothalamic sites microdissected from fresh brains can be assessed in a reliable fashion and are in accord with the proposal that enhanced NPY action within the PVN is responsible for increased drive for food.
- Published
- 1992
- Full Text
- View/download PDF
44. Involvement of the Y-1 receptor subtype in the regulation of luteinizing hormone secretion by neuropeptide Y in rats.
- Author
-
Kalra SP, Fuentes M, Fournier A, Parker SL, and Crowley WR
- Subjects
- Analysis of Variance, Animals, Calcium pharmacology, Cells, Cultured, Cerebral Ventricles metabolism, Estradiol pharmacology, Female, Indomethacin pharmacology, Injections, Intraventricular, Luteinizing Hormone blood, Median Eminence drug effects, Median Eminence metabolism, Methoxamine pharmacology, Neuropeptide Y administration & dosage, Neuropeptide Y metabolism, Ovariectomy, Progesterone pharmacology, Rats, Rats, Inbred Strains, Receptors, Neuropeptide Y, Receptors, Neurotransmitter drug effects, Ryanodine pharmacology, Cerebral Ventricles physiology, Gonadotropin-Releasing Hormone metabolism, Luteinizing Hormone metabolism, Median Eminence physiology, Neuropeptide Y pharmacology, Receptors, Neurotransmitter physiology
- Abstract
The present experiments were designed to investigate structure-function relationships, and identify the receptor subtype and postreceptor cellular mechanisms that mediate the ovarian hormone-dependent, excitatory, and inhibitory effects of neuropeptide Y (NPY) on LH release in female rats. Intracerebroventricular administration of NPY decreased plasma concentrations of LH in ovariectomized, hormonally untreated rats but stimulated LH release in ovariectomized rats pretreated with estradiol benzoate and progesterone. A similar dual response was also obtained after administration of NPY2-36. However, deletion of additional amino acids at the N terminus, as in NPY5-36, NPY11-36, NPY16-36, and NPY25-36, rendered the peptides inactive. An N-terminal fragment, NPY1-24-amide, and a discontinuous NPY analog, NPY1-4-epsilon-amino-caproic acid-25-36, similarly failed to influence LH release. The analog [Leu31,Pro34]NPY, a preferential agonist at the Y-1 NPY receptor subtype, also elicited the dual LH responses, but the preferential Y-2 receptor agonist, NPY13-36, was completely inactive. Further, only the peptides that stimulated LH release in vivo, i.e. NPY, NPY2-36, and [Leu31,Pro34]NPY, also stimulated the release of LHRH from median eminence fragments of steroid-primed rats in vitro, and the excitatory effect of [Leu31,Pro34] NPY was blocked by a noncompetitive NPY receptor antagonist, D-myo-inositol-1,2,6-trisphosphate. With the exception of NPY1-24-amide, all of the NPY fragments tested bound specifically to NPY binding sites in hypothalamic membrane preparations, but the highest binding affinities were found for the peptides that evoked biological responses in the in vivo and in vitro tests. Further analysis of the mode of action of NPY showed that the stimulation of LHRH release in vitro was unaffected by omission of Ca2+ from the incubation medium, but was prevented by two antagonists of intracellular Ca2+ mobilization, 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester and ryanodine. Inhibition of prostaglandin synthesis with indomethacin blocked the stimulatory effect of the alpha 1-adrenergic agonist methoxamine on LHRH release, but not the increase produced by NPY.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1992
- Full Text
- View/download PDF
45. Steroidal regulation of hypothalamic neuropeptide Y release and gene expression.
- Author
-
Sahu A, Phelps CP, White JD, Crowley WR, Kalra SP, and Kalra PS
- Subjects
- Animals, Gene Expression drug effects, Hypothalamus drug effects, Hypothalamus metabolism, Male, Neuropeptides genetics, Orchiectomy, RNA genetics, RNA isolation & purification, RNA, Messenger genetics, RNA, Messenger isolation & purification, Rats, Rats, Inbred Strains, Reference Values, Hypothalamus physiology, Neuropeptide Y genetics, Neuropeptides metabolism, Protein Precursors genetics, RNA, Messenger metabolism, Testis physiology, Testosterone pharmacology
- Abstract
Neuropeptide Y (NPY) readily stimulates the release of hypothalamic LHRH and pituitary LH release in intact and gonadal steroid-primed gonadectomized rats. We have now tested the hypothesis that the release and synthesis of hypothalamic NPY may be regulated by gonadal steroids. To measure the effects of gonadal hormones on NPY release, a permanent push-pull cannula was implanted in the anterior pituitary (AP) of sham castrated (controls) or castrated (CAST) male rats, and 1 week later, the AP was perfused with artificial cerebrospinal fluid over a 3-4 h period. NPY concentrations in the perfusates collected at 10-min intervals were measured by RIAs. The NPY release pattern in the AP was episodic in both intact and CAST rats, and the frequency of NPY episodes was similar in two groups. However, the amount of NPY detected in the AP of CAST rats was significantly less than that of intact rats because the mean rate of release and the amplitude of NPY episodes in the perfusates of CAST rats were significantly reduced. This observation of attenuated hypothalamic NPY output in vivo and previous evidence of decreased hypothalamic NPY contents after CAST implied that the synthesis of hypothalamic NPY may be regulated by testicular secretions. Therefore, the effects of testosterone (T)-replacement on preproNPY messenger RNA (mRNA) in the medial basal hypothalamus (MBH) was evaluated. Rats were CAST and received either empty or T-filled Silastic capsules sc. Two weeks later, the level of perproNPY mRNA in the MBH was determined by solution hybridization/ribonuclease protection assay using a complementary RNA probe complementary to the rat NPY precursor mRNA. We observed that the levels of preproNPY mRNA were 2-fold higher in the MBH of T-replaced CAST as compared to control CAST rats. These findings are consistent with the hypothesis that gonadal steroids enhance the neurosecretory activity of hypothalamic NPYergic neurons, and for the first time reveal a coupling between the level of gene expression and the secretion of a neuropeptide involved in the regulation of hypothalamic LHRH and pituitary LH release.
- Published
- 1992
- Full Text
- View/download PDF
46. Effects of tachykinins on luteinizing hormone release in female rats: potent inhibitory action of neuropeptide K.
- Author
-
Sahu A and Kalra SP
- Subjects
- Animals, Dose-Response Relationship, Drug, Estradiol analogs & derivatives, Estradiol pharmacology, Female, Gonadotropin-Releasing Hormone metabolism, Injections, Intraventricular, Neurokinin A pharmacology, Ovariectomy, Pituitary Gland drug effects, Pituitary Gland metabolism, Potassium pharmacology, Progesterone pharmacology, Radioimmunoassay, Rats, Substance P pharmacology, Tachykinins administration & dosage, Time Factors, Luteinizing Hormone blood, Neuropeptides pharmacology, Tachykinins pharmacology
- Abstract
Tachykinins, a family of biologically active related peptides, are found in variable amounts in the rat hypothalamus. We assessed the effects of five tachykinins, substance P (SP), neurokinin A (NKA), neuropeptide K (NPK), neuropeptide gamma (NP gamma), and neurokinin B (NKB), on LH release in different experimental model systems in ovariectomized rats. In the first series of experiments rats were ovariectomized and implanted with permanent cannulae in the third cerebroventricle of the rat brain. Two weeks later, the effects of intracerebroventricular injection of 0.5 or 1.25 nm various tachykinins on LH release were studied. The results showed that whereas SP, NKA, and NKB were ineffective, and NP gamma was marginally effective, NPK produced a long-lasting suppression of LH release. NPK decreased LH release in a dose- and time-related fashion. Similarly, in the second series of experiments, whereas SP and NKA were inactive, NPK completely suppressed the LH surge induced by progesterone in estrogen-primed ovariectomized rats. In the third series of experiments we observed that NK-2 receptor agonist [Nle10]NKA4-10, and not NK-1 receptor agonist [Sar9,Met(O2)11]SP, suppressed both the release of LH in vivo and basal and KCl-induced hypothalamic LHRH release in vitro. These results show that NPK is the most effective tachykinin in suppressing LH release, and the inhibitory response is mediated by hypothalamic NK-2 receptors. These findings are in accord with the hypothesis that NPK may serve as a hypothalamic inhibitory neurotransmitter/neuromodulator of LHRH secretion.
- Published
- 1992
- Full Text
- View/download PDF
47. Neuropeptide Y modulates the binding of a gonadotropin-releasing hormone (GnRH) analog to anterior pituitary GnRH receptor sites.
- Author
-
Parker SL, Kalra SP, and Crowley WR
- Subjects
- Animals, Female, Gastrointestinal Hormones metabolism, Magnesium Chloride pharmacology, Peptide YY, Peptides metabolism, Rats, Rats, Inbred Strains, Gonadotropin-Releasing Hormone analogs & derivatives, Neuropeptide Y physiology, Pituitary Gland, Anterior metabolism, Receptors, LHRH metabolism
- Abstract
Neuropeptide Y (NPY) increases LH secretion in part by enhancing the release of LH in response to GnRH. The present studies examined whether NPY influences the binding of GnRH to its receptors and also assessed whether specific binding sites for NPY exist in rat anterior pituitary membranes. In concentrations from 66-200 nM, NPY dose-dependently enhanced the binding of a 125I-labeled GnRH agonist, [D-Ala6, des-Gly10]GnRH ethylamide (GnRHa; 30 pM) to anterior pituitary membranes of chronically ovariectomized rats; higher concentrations of NPY were ineffective. At 200 nM, NPY significantly increased the high affinity binding of the GnRHa to these membranes, without change in the apparent maximum binding capacity. Further, 200 nM NPY significantly increased the binding of [125I]GnRHa when this tracer was used in concentrations of less than 100 pM, but NPY did not increase the binding of higher concentrations of [125I]GnRHa. Peptide YY, a gastrointestinal peptide structurally and functionally related to NPY, and the hypothalamic/pituitary peptide galanin (both at 200 nM) also increased the binding of [125I]GnRHa (30 pM) to anterior pituitary membranes, but to approximately one third of the extent produced by NPY. Salmon calcitonin, which, similar to NPY, is a strongly hydrophobic neuroendocrine peptide, did not alter GnRHa binding. Mg++ ion dose-dependently reduced the affinity of GnRHa binding, without changing the maximum binding capacity, and also attenuated the increase in GnRHa binding produced by NPY. To further characterize the nature of NPY interaction with anterior pituitary membranes, [125I] peptide YY was used to label NPY binding sites in anterior pituitary and hypothalamic membranes from ovariectomized rats. Specific, and predominantly high affinity, NPY binding sites were demonstrated in hypothalamic membranes, whereas NPY binding to anterior pituitary membranes could be resolved into high and low affinity components. These results show that at low nanomolar concentrations, NPY can enhance the association of GnRHa to receptors in anterior pituitary membranes. This demonstration of increased GnRHa binding in the presence of NPY may explain in part the action of this neuropeptide to potentiate GnRH-induced LH release from anterior pituitary cells.
- Published
- 1991
- Full Text
- View/download PDF
48. Localization of neuropeptide-Y immunoreactivity in estradiol-concentrating cells in the hypothalamus.
- Author
-
Sar M, Sahu A, Crowley WR, and Kalra SP
- Subjects
- Animals, Autoradiography, Female, Hypothalamus cytology, Immunoenzyme Techniques, Kinetics, Neuropeptide Y analysis, Rats, Rats, Inbred Strains, Tritium, Estradiol metabolism, Hypothalamus metabolism, Neuropeptide Y metabolism
- Abstract
Considerable evidence shows that gonadal steroids exert a facilitatory influence on levels and release of neuropeptide-Y (NPY) from the hypothalamus. However, it is not known whether gonadal steroids act directly on NPY-producing cells in the arcuate nucleus (ARC) of the hypothalamus to produce these facilitatory effects on NPY or whether they act on other cells that have a modulatory influence via synapses on ARC NPY cells. We applied the combined method of steroid autoradiography and immunocytochemistry to assess the localization of [3H]estradiol in relation to NPY-producing cells in the hypothalamus. Rats (n = 6) were bilaterally ovariectomized and injected intracerebroventricularly with colchicine. Twenty-four hours later each rat received an iv injection of 17 beta-[2,4,6,7,16,17(-3)H]estradiol (SA, 166 Ci/mmol) at a dose of 5.0 micrograms/kg BW. One hour after the injection of [3H]estradiol, the rats were perfused with 4% paraformaldehyde; brains were removed, frozen in isopentane precooled in liquid nitrogen (-190 C), sectioned, and processed for autoradiography. The autoradiograms were then incubated with specific antibodies for NPY immunostaining by the avidin-biotin-peroxidase method. The results revealed NPY-immunopositive cells in the ARC, striatum, hippocampus, amygdala, and cerebral cortex and a few cells in the median eminence. NPY-immunoreactive fibers were also detected in the internal layer of the median eminence. The largest number of neurons showing NPY immunoreactivity in the cytoplasm was detected in the ARC, and only in this nucleus did we observed colocalization of [3H]estradiol and NPY immunoreactivity in neurons. A population of NPY-immunopositive cells in the ARC (10-20%) exhibited nuclear [3H]estradiol; the majority of these cells were located in the lateral and ventral portions of the ARC. Since gonadal steroids stimulate hypothalamic NPY levels and release, this anatomical evidence of colocalization is suggestive of a direct genomic modulation of NPY neurosecretion by steroids in a subpopulation of hypothalamic NPY-immunopositive neurons.
- Published
- 1990
- Full Text
- View/download PDF
49. Endogenous opioid peptides mediate the interleukin-1-induced inhibition of the release of luteinizing hormone (LH)-releasing hormone and LH.
- Author
-
Kalra PS, Fuentes M, Sahu A, and Kalra SP
- Subjects
- Animals, Corticosterone blood, Female, Gonadotropin-Releasing Hormone metabolism, Hypothalamus metabolism, Injections, Intraventricular, Luteinizing Hormone metabolism, Naloxone pharmacology, Ovariectomy, Rats, Steroids, Endorphins physiology, Gonadotropin-Releasing Hormone antagonists & inhibitors, Interleukin-1 pharmacology, Luteinizing Hormone antagonists & inhibitors
- Abstract
We have reported recently that central administration of both the alpha- and beta-subtypes of the cytokine interleukin-1 (IL-1) inhibited the estrogen-progesterone-induced LH surge in ovariectomized (ovx) rats. This inhibition was probably due to a central effect, since IL-1 alpha and IL-1 beta also suppressed the in vitro LHRH output from the hypothalami of steroid-primed ovx rats. Whether IL-1 inhibits LHRH release by a direct action or via some other neuronal system is not known. Since IL-1 reportedly stimulates the release of POMC peptides, which are known to be inhibitory to the LHRH-LH axis, we have tested the hypothesis that the inhibitory influence of IL-1 may be mediated via activation of hypothalamic opioid peptides. Ovx rats, preimplanted with cannulae in the third ventricle of the brain, were injected with 30 micrograms estradiol benzoate, followed by 2 mg progesterone 48 h later. Three hours after P injection, IL-1 alpha, IL-1 beta, or saline (SAL) was injected intracerebroventricularly (30 ng/3 microliters) at 1300 h, followed immediately by iv infusion of SAL or the opiate antagonist naloxone hydrochloride (NAL; 2 mg/0.6 ml.h) for 2 h. Plasma LH levels were measured in blood samples withdrawn hourly until 1800 h. Both IL-1 alpha and IL-1 beta blocked the afternoon LH surge. NAL infusion into control SAL-injected rats did not alter the LH surge; however, it reversed the IL-1 alpha- and IL-1 beta-induced suppression of the LH surge. To determine whether this reversal of IL-1 suppression of the LH surge was due to NAL action at the hypothalamic level, the preoptic area-medial basal hypothalamus of similarly primed ovx rats was obtained at 1300 h and incubated in vitro in the presence of 10 nM IL-1 alpha or IL-1 beta with or without 100 micrograms/ml NAL. Both subtypes of IL-1 suppressed LHRH output significantly. NAL alone did not affect LHRH release, but it completely reversed the inhibitory effects of the cytokine on LHRH release. These results suggest that IL-1 alpha and IL-1 beta inhibit LHRH-LH release by stimulating the activity of hypothalamic endogenous opioid peptide systems.
- Published
- 1990
- Full Text
- View/download PDF
50. Functional heterogeneity in neuropeptide-Y-producing cells in the rat brain as revealed by testosterone action.
- Author
-
Sahu A, Kalra PS, Crowley WR, and Kalra SP
- Subjects
- Animals, Brain cytology, Denervation, Hypothalamus metabolism, Luteinizing Hormone blood, Male, Orchiectomy, Rats, Reference Values, Testosterone blood, Brain metabolism, Neuropeptide Y metabolism, Testosterone pharmacology
- Abstract
Despite the widespread distribution of neuropeptide-Y (NPY) in various hypothalamic sites, castration reduced and testosterone (T) replacement restored NPY levels selectively in the median eminence (ME), arcuate nucleus (ARC), and ventromedial nucleus (VMN). Since androgen-concentrating cells and NPY-producing cells display overlapping distribution in the ARC and brain stem (BS), we assessed the participation of BS NPY cells in the steroid-dependent site-specific effects on NPY levels. The BS projections to the hypothalamus were severed by bilateral neural transection (BNT) with a knife lowered on either side of the sagittal sinus to the depth of the dorsal tegmentum in the mesencephalon in intact, castrated, or castrated rats that additionally received sc T implants to maintain physiological T levels. Two weeks later, NPY concentrations in microdissected hypothalamic sites and serum LH and T levels were quantitated by RIA. Castration decreased and T replacement increased NPY concentrations in only three sites, such as the VMN, ARC, and ME. In response to BNT in gonadally intact rats, a different site-specific response of NPY levels was observed; NPY levels decreased in the ME, as seen after castration, and, additionally, decreased in the medial preoptic area, paraventricular nucleus, and dorsomedial nucleus, suggesting that BS NPY neurons innervate these four sites. When castration and BNT were performed simultaneously, a combined regional response was evident. NPY levels decreased in six sites, including two sites (ARC and VMN) that normally respond to castration alone, three sites (medial preoptic area, paraventricular nucleus, and dorsomedial nucleus) that normally respond to BNT alone, and the ME, the only site that showed reduction of NPY levels of similar magnitude after either castration or BNT, but the response of combined surgery was not additive. This observation suggested that gonadal steroids act outside the hypothalamus to raise ME NPY levels, and therefore, BNT in intact rats impaired the effectiveness of steroids. To test this hypothesis, the effects of physiological T replacement in the castrate plus BNT group were studied. We observed that whereas T replacement readily raised NPY levels in the VMN and ARC, it was completely ineffective in the third T-dependent site, the ME. Collectively, these findings revealed a functional heterogeneity among NPY-producing cells in response to T. Apparently, there are two distinct neural sites in the rat brain where T acts to exert a site-specific stimulatory effect on NPY in the hypothalamus.(ABSTRACT TRUNCATED AT 400 WORDS)
- Published
- 1990
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.