354 results on '"Dj, Gilbert"'
Search Results
2. Elk-L3, a novel transmembrane ligand for the Eph family of receptor tyrosine kinases, expressed in embryonic floor plate, roof plate and hindbrain segments
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Nw, Gale, Flenniken A, Dc, Compton, Jenkins N, Ng, Copeland, Dj, Gilbert, Davis S, David Wilkinson, and Gd, Yancopoulos
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Male ,DNA, Complementary ,Base Sequence ,Sequence Homology, Amino Acid ,Molecular Sequence Data ,Chromosome Mapping ,Proteins ,Receptor Protein-Tyrosine Kinases ,Ephrin-B1 ,Nervous System ,Rats ,Mice, Inbred C57BL ,Rhombencephalon ,Mice ,Animals ,Humans ,Female ,Amino Acid Sequence ,Cloning, Molecular ,Chromosomes, Human, Pair 17 - Abstract
The Eph family of receptor tyrosine kinases has 13 distinct members and seven ligands for these receptors have been described to date. These receptors and their ligands have been implicated in regulating neuronal axon guidance and in patterning of the developing nervous system and may also serve a patterning and compartmentalization role outside of the nervous system as well. The ligands are all membrane-attached, and this attachment appears to be crucial for their normal function; five of the known ligands are linked to the membrane via a glycosyl phosphotidylinositol (GPI) linkage, while two of the ligands are transmembrane proteins. Despite the large number of Eph family receptors and ligands, they can be divided into just two major subclasses based on their binding specificities. All the GPI-anchored ligands bind and activate one subclass of the Eph receptors (that represented by Eck) while the two transmembrane ligands bind and activate the other major subclass of receptors (represented by Elk). Here we report the identification and characterization of the third, and most divergent, member of the transmembrane group of Eph ligands, which we term Elk-L3 (Elk-related receptor ligand number 3). Elk-L3 is notable for its remarkably restricted and prominent expression in the floor plate and roof plate of the developing neural tube and its rhombomere-specific expression in the developing hindbrain. The Elk-L3 gene has been localized to mouse chromosome 11 and human chromosome 17.
- Published
- 1996
3. Four novel members of the connexin family of gap junction proteins. Molecular cloning, expression, and chromosome mapping
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Jacques-Antoine Haefliger, Bruzzone R, Na, Jenkins, Dj, Gilbert, Ng, Copeland, and Dl, Paul
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Genomic Library ,Base Sequence ,Protein Conformation ,Molecular Sequence Data ,Chromosome Mapping ,Gene Expression ,Membrane Proteins ,Rats, Inbred Strains ,DNA ,Blotting, Northern ,Polymerase Chain Reaction ,Connexins ,Rats ,Models, Structural ,Blotting, Southern ,Oligodeoxyribonucleotides ,Multigene Family ,Sequence Homology, Nucleic Acid ,Animals ,RNA ,Amino Acid Sequence ,Cloning, Molecular ,Crosses, Genetic - Abstract
We have used low stringency hybridization and polymerase chain reaction (PCR) amplification with degenerate oligonucleotides to identify four new members of the rat connexin gene family. On the basis of their predicted molecular mass, these proteins have been designated connexin (Cx) 40 (Cx40), Cx37, Cx33, and Cx31.1. The new connexins exhibit all of the conserved structural features of the connexin family, including highly similar extracellular and transmembrane domains but divergent major cytoplasmic domains. On the basis of primary sequence similarity, the connexin family may be divided into two classes. Cx40, Cx37, and Cx33 are similar to the previously characterized Cx43 and Cx46. Cx31.1 is similar to Cx26, Cx31, and Cx32. Cx37 and Cx40 mRNAs are expressed in a wide variety of adult organs and tissues, with particular abundance in lung. However, their relative levels are different in many organs and thus their distribution is not completely coincident. Cx33 and Cx31.1 genes exhibit a much more restricted pattern of expression; mRNAs are detected only in testes and skin, respectively. Chromosomal mapping studies indicate that Cx26 and Cx46 are tightly linked on chromosome 14, and Cx37 and Cx31.1 are linked on chromosome 4, while the rest of the connexin genes are dispersed.
- Published
- 1992
4. Cloning, expression, and chromosomal localization of beta-adrenergic receptor kinase 2. A new member of the receptor kinase family
- Author
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Jl, Benovic, Jj, Onorato, Jl, Arriza, Wc, Stone, Martin Lohse, Na, Jenkins, Dj, Gilbert, Ng, Copeland, Mg, Caron, and Rj, Lefkowitz
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G-Protein-Coupled Receptor Kinase 3 ,Base Sequence ,Molecular Sequence Data ,Chromosome Mapping ,DNA ,Blotting, Northern ,Cyclic AMP-Dependent Protein Kinases ,Mice ,beta-Adrenergic Receptor Kinases ,Cricetinae ,Animals ,Cattle ,Amino Acid Sequence ,RNA, Messenger ,Cloning, Molecular ,Protein Kinases - Abstract
The beta-adrenergic receptor kinase (beta ARK) specifically phosphorylates the agonist-occupied form of the beta-adrenergic and related G protein-coupled receptors. Structural features of this enzyme have been elucidated recently by the isolation of a cDNA that encodes bovine beta ARK. Utilizing a catalytic domain fragment of the beta ARK cDNA to screen a bovine brain cDNA library we have isolated a clone encoding a beta ARK-related enzyme which we have termed beta ARK2. Overall, this enzyme has 85% amino acid identity with beta ARK, with the protein kinase catalytic domain having 95% identity. The ability of beta ARK2 to phosphorylate various substrates was studied after expression in COS 7 cells. Although beta ARK2 is essentially equiactive with beta ARK in phosphorylating an acid-rich synthetic model peptide it was only approximately 50% as active when the substrate was the agonist-occupied beta 2-adrenergic receptor and only approximately 20% as active toward light-bleached rhodopsin. As with beta ARK, phosphorylation of the receptor substrates by beta ARK2 was completely stimulus dependent. RNA blot analysis with selected bovine tissues reveals an mRNA of 8 kilobases with a distribution similar to that of beta ARK. More detailed RNA analysis using a ribonuclease protection assay in various rat tissues suggests that the beta ARK2 message is present at much lower levels (typically 10-20%) than the beta ARK message. In the rat the beta ARK2 mRNA is localized predominantly in neuronal tissues although low levels are also observed in various peripheral tissues. The beta ARK2 gene has been localized to a region of mouse chromosome 5 whereas the beta ARK gene is localized on mouse chromosome 19. These data suggest the existence of a "family" of receptor kinases which may serve broadly to regulate receptor function.
- Published
- 1991
5. Matrix glycoprotein SC1/ECM2 augments B lymphopoiesis
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Oritani K, Kanakura Y, Aoyama K, Yokota T, Ng, Copeland, Dj, Gilbert, Na, Jenkins, Yoshiaki Tomiyama, Matsuzawa Y, and Pw, Kincade
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B-Lymphocytes ,Extracellular Matrix Proteins ,Recombinant Fusion Proteins ,Molecular Sequence Data ,Chromosome Mapping ,Immunoglobulins ,Cell Differentiation ,Nerve Tissue Proteins ,Hematopoiesis ,Rats ,Mice ,Activated-Leukocyte Cell Adhesion Molecule ,Animals ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Stromal Cells ,Sequence Alignment - Abstract
The extracellular matrix produced by stromal cells plays a critical role in lympho-hematopoiesis. It was recently discovered that matrix glycoprotein SC1/ECM2 is a component of that matrix and preliminary evidence suggested that it could contribute to the nurturing environment for B-lymphocyte precursors. A fusion protein prepared from the amino terminal portion of SC1/ECM2 and the constant region of human Ig preferentially bound to pre-B cells. Furthermore, the cloning efficiency of interleukin-7-dependent B-cell precursors was increased in a dose-dependent manner by addition of this fusion protein. We now report the complete cDNA sequence for murine SC1/ECM2 and its localization to the central region of chromosome 5. A fusion protein prepared from the full length of SC1/ECM2 and Ig was found to recognize pre-B cells in a divalent cation-dependent manner, and to augment mitogen-dependent proliferation of mature B cells, as well as the cloning of pre-B cells, but to have no influence on myeloid progenitor cells. Although SC1/ECM2 is normally a secreted protein, we show that it is also capable of augmenting lymphopoiesis when expressed as a transmembrane protein on fibroblasts. Although the C-terminal portion of SC1/ECM2 has sequence homology to osteonectin/SPARC, the unique N-terminal one fifth of the protein was sufficient to augment lymphocyte growth.
6. Genomic organization and chromosome location of the murine Rpl23 gene
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Natascha Kleiter, Artner I, Ng, Copeland, Dj, Gilbert, Na, Jenkins, and Kratochwil K
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Male ,Ribosomal Proteins ,Mice, Inbred BALB C ,Genetic Linkage ,Escherichia coli Proteins ,Molecular Sequence Data ,Chromosome Mapping ,Exons ,Physical Chromosome Mapping ,Chromosomes ,Introns ,Mice, Inbred C57BL ,Mice ,Animals ,Humans ,Female ,RNA, Messenger ,Cloning, Molecular ,Crosses, Genetic - Abstract
The mouse gene coding for ribosomal protein L23 (Rpl23) has been fully sequenced, including 580 bp of the 5' upstream region. The 5-kb gene comprises 5 exons and contains an unusually long (3,153 bp) third intron. The gene was mapped to the distal region of mouse chromosome 11, homologous to human chromosome 17q21--q22.
7. Foetal Alcohol Spectrum Disorder (FASD) and the Courts: How England and Wales Could Benefit From Following an Australian Model.
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Hill G, Gerry Kc F, Herlihen P, Allely CS, and Gilbert DJ
- Abstract
Foetal alcohol spectrum disorder (FASD) is a set of symptoms and signs that may follow from exposure of the unborn child to alcohol during pregnancy. Characterised by cognitive and behavioural impairments, one secondary outcome from FASD, is encounters with the criminal justice system (CJS). In some countries, for example, England and Wales, it seems likely that many cases are missed at this point and, thus, courts are at risk of making unsafe judgements. We could learn a lot from countries where services are generally more used to dealing with FASD. Australia is one such country., (© 2025 John Wiley & Sons Ltd.)
- Published
- 2025
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8. Fetal alcohol spectrum disorder (FASD) and the criminal justice system: A guide for legal professionals.
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Brown J, Lewis DS, Kivisalu T, Wartnik AP, Carter MN, Harr D, Jozan A, and Gilbert DJ
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- Humans, Female, Pregnancy, Fetal Alcohol Spectrum Disorders psychology, Criminal Law legislation & jurisprudence
- Abstract
Fetal Alcohol Spectrum Disorder (FASD) is a lifelong disorder caused by prenatal alcohol exposure (PAE) and is one of the most common causes of brain damage and developmental disability. FASD has been characterized by an array of symptoms that negatively affects cognitive, social, and adaptive functioning. Individuals living with FASD, relative to the general population, are more likely to become entangled in the legal system due to challenges associated with the disorder. Moreover, symptomology of FASD often contributes to these individuals struggling in successfully navigating various stages of the legal system, including arrest, interrogation, working with their defense counsel, and courtroom appearances. The difficulties faced by defendants living with FASD are exacerbated by systemic failure from legal professionals in recognizing and accommodating for the extent and complexities of this disorder. Consequently, defendants living with FASD often do not receive effective due process of law in comparison to their neurotypical peers. Moreover, attempts at punishment and deterrence through probation and jail terms are often ineffective for individuals living with FASD because of the effects of their disorder. This article is intended to provide valuable information and best practices for professionals in the legal system, particularly judges, prosecutors, defense attorneys, social workers/mitigation specialists, and psychologists, who are likely to encounter individuals living with FASD or suspected FASD early in the judicial process., Competing Interests: Declaration of competing interest None., (Copyright © 2024. Published by Elsevier Ltd.)
- Published
- 2024
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9. Application of Reproductive Technologies to the Critically Endangered Baw Baw Frog, Philoria frosti .
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Silla AJ, Hobbs RJ, Gilbert DJ, Goodall D, Parrott ML, Lee A, O'Brien JK, and Byrne PG
- Abstract
Reproductive technologies (RTs) can assist integrated conservation breeding programs to attain propagation targets and manage genetic diversity more effectively. While the application of RTs to enhance the conservation management of threatened amphibians has lagged behind that of other taxonomic groups, a recent surge in research is narrowing the divide. The present study reports on the first application of RTs (hormone-induced spawning, hormone-induced sperm-release, and sperm cryopreservation) to the critically endangered Baw Baw frog, Philoria frosti . To determine the effect of hormone therapy on spawning success, male-female pairs were administered either 0 μg/g gonadotropin-releasing hormone agonist (GnRHa), 0.5 μg/g GnRHa, or 0.5 μg/g GnRHa + 10 μg/g metoclopramide (MET) ( n = 6-7 pairs/treatment), and the number of pairs ovipositing, total eggs, and percent fertilisation success were quantified. To determine the effect of hormone therapy on sperm-release and to establish the peak time to collect sperm post-hormone administration, males were administered 0 IU/g ( n = 4), or 20 IU/g hCG ( n = 16). Total sperm, sperm concentration, and percent viability were quantified at 0, 2, 4, 6, 8, 10, and 12 h post-hormone administration. Overall, the percentage of pairs ovipositing was highest in the GnRHa + MET treatment, with 71% of pairs ovipositing, compared to 57% and 33% of pairs in the GnRHa and control treatments, respectively. The quantity of sperm released from males in response to hCG peaked at 4 h post-hormone administration, though it remained high up to 12 h. The percent sperm viability also peaked at 4 h post-administration (94.5%), exhibiting a steady decline thereafter, though viability remained above 77% throughout the 12 h collection period. The remaining sperm samples ( n = 22) were cryopreserved using established protocols and biobanked for long-term storage and future conservation applications. The mean post-thaw sperm viability was 59%, and the percent total motility was 17%. The results from this preliminary study will direct further applications of RTs to the critically endangered Baw Baw frog to assist with species recovery.
- Published
- 2023
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10. Foetal alcohol spectrum disorder and Investigative interviewing: A systematic review highlighting clinical and legal implications and recommendations.
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Gilbert DJ, Allely CS, Mukherjee RAS, and Cook PA
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- Crime, Female, Humans, Narration, Pregnancy, Suggestion, Fetal Alcohol Spectrum Disorders diagnosis, Fetal Alcohol Spectrum Disorders epidemiology
- Abstract
Individuals with foetal alcohol spectrum disorders (FASD) are estimated to be 19 times more likely to encounter the criminal justice system (CJS) in comparison to individuals without FASD. During encounters with the CJS, investigative interviews are employed to obtain accurate information from suspects, victims or witnesses of crime. A systematic search using PRISMA guidelines was performed to identify empirical studies published that have explored the questioning of the FASD population within the CJS and the vulnerabilities of FASD-impacted individuals during investigative interviewing. A total of 383 studies were identified from the databases searched and 7 further studies were identified from Google Scholar. After deduplication, abstract and title screening, the full text of 23 studies were assessed for inclusion and 5 were included in the narrative synthesis of results. Two papers were empirical studies focussed on the performance of FASD-impacted individuals during investigative interviewing. Whilst the first study found the FASD population susceptible to suggestions, the second (a case study), identified the ploys employed during investigative interviewing to obtain a confession. Three papers studied the wider vulnerabilities of FASD-impacted individuals and found diminished psycho-legal abilities, increased risk of recidivism and biological, psychological and social factors that render FASD-impacted individuals vulnerable to CJS encounters. Despite the greater likelihood of CJS encounters, the result of this review highlights the slim evidence base useful to establish the vulnerabilities of FASD-impacted individuals within the CJS., (© 2021 John Wiley & Sons Ltd.)
- Published
- 2022
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11. T Cell-Mediated Antitumor Immunity Cooperatively Induced By TGFβR1 Antagonism and Gemcitabine Counteracts Reformation of the Stromal Barrier in Pancreatic Cancer.
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Li D, Schaub N, Guerin TM, Bapiro TE, Richards FM, Chen V, Talsania K, Kumar P, Gilbert DJ, Schlomer JJ, Kim SJ, Sorber R, Teper Y, Bautista W, Palena C, Ock CY, Jodrell DI, Pate N, Mehta M, Zhao Y, Kozlov S, and Rudloff U
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- Animals, Antimetabolites, Antineoplastic pharmacology, Apoptosis, CD8-Positive T-Lymphocytes drug effects, CD8-Positive T-Lymphocytes metabolism, Carcinoma, Pancreatic Ductal drug therapy, Carcinoma, Pancreatic Ductal metabolism, Carcinoma, Pancreatic Ductal pathology, Cell Proliferation, Combined Modality Therapy, Deoxycytidine pharmacology, Humans, Mice, Pancreatic Neoplasms drug therapy, Pancreatic Neoplasms metabolism, Pancreatic Neoplasms pathology, Stromal Cells drug effects, Stromal Cells metabolism, Tumor Cells, Cultured, Xenograft Model Antitumor Assays, Gemcitabine, CD8-Positive T-Lymphocytes immunology, Carcinoma, Pancreatic Ductal immunology, Deoxycytidine analogs & derivatives, Pancreatic Neoplasms immunology, Receptor, Transforming Growth Factor-beta Type I antagonists & inhibitors, Stromal Cells immunology, Tumor Microenvironment
- Abstract
The desmoplastic stroma of pancreatic cancers forms a physical barrier that impedes intratumoral drug delivery. Attempts to modulate the desmoplastic stroma to increase delivery of administered chemotherapy have not shown positive clinical results thus far, and preclinical reports in which chemotherapeutic drugs were coadministered with antistromal therapies did not universally demonstrate increased genotoxicity despite increased intratumoral drug levels. In this study, we tested whether TGFβ antagonism can break the stromal barrier, enhance perfusion and tumoral drug delivery, and interrogated cellular and molecular mechanisms by which the tumor prevents synergism with coadministered gemcitabine. TGFβ inhibition in genetically engineered murine models (GEMM) of pancreas cancer enhanced tumoral perfusion and increased intratumoral gemcitabine levels. However, tumors rapidly adapted to TGFβ-dependent stromal modulation, and intratumoral perfusion returned to pre-treatment levels upon extended TGFβ inhibition. Perfusion was governed by the phenotypic identity and distribution of cancer-associated fibroblasts (CAF) with the myelofibroblastic phenotype (myCAFs), and myCAFs which harbored unique genomic signatures rapidly escaped the restricting effects of TGFβ inhibition. Despite the reformation of the stromal barrier and reversal of initially increased intratumoral exposure levels, TGFβ inhibition in cooperation with gemcitabine effectively suppressed tumor growth via cooperative reprogramming of T regulatory cells and stimulation of CD8 T cell-mediated antitumor activity. The antitumor activity was further improved by the addition of anti-PD-L1 immune checkpoint blockade to offset adaptive PD-L1 upregulation induced by TGFβ inhibition. These findings support the development of combined antistroma anticancer therapies capable of impacting the tumor beyond the disruption of the desmoplastic stroma as a physical barrier to improve drug delivery., (©2021 The Authors; Published by the American Association for Cancer Research.)
- Published
- 2021
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12. Warmer temperature and provision of natural substrate enable earlier metamorphosis in the critically endangered Baw Baw frog.
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Gilbert DJ, Magrath MJL, and Byrne PG
- Abstract
Temperature and food availability are known to independently trigger phenotypic change in ectotherms, but the interactive effects between these factors have rarely been considered. This study investigates the independent and interactive effects of water temperature and food availability on larval growth and development of the critically endangered Baw Baw frog, Philoria frosti . Larvae were reared at low (12°C) or high (17°C) water temperature in the absence or presence of substrate that controlled food availability, and body size and time to metamorphosis were quantified. Growth and development of larvae was influenced by the individual effects of temperature and food availability; time to metamorphosis was shorter in warm water treatment groups and in the presence of substrate and increased food. Unexpectedly, however, water temperature and food availability did not have an interactive effect on either time to metamorphose or body size at metamorphosis. Under all treatment groups, metamorphic onset occurred once a developmental size threshold was reached, indicating that growth rate and body size are key factors controlling the metamorphic process in Baw Baw frogs (consistent with the Wilbur-Collins model for ectotherm development). From an applied perspective, our findings have implications for amphibian conservation because they indicate that simple manipulations of temperature and food availability can be used to increase the rate of frog production in conservation breeding programs., (© The Author(s) 2020. Published by Oxford University Press and the Society for Experimental Biology.)
- Published
- 2020
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13. Decline in atmospheric sulphur deposition and changes in climate are the major drivers of long-term change in grassland plant communities in Scotland.
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Mitchell RJ, Hewison RL, Fielding DA, Fisher JM, Gilbert DJ, Hurskainen S, Pakeman RJ, Potts JM, and Riach D
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- Bryophyta, Ecosystem, Nitrogen analysis, Poaceae classification, Poaceae growth & development, Scotland, Sulfur analysis, Biodiversity, Climate Change, Grassland, Plants classification, Sulfur pharmacology
- Abstract
The predicted long lag time between a decrease in atmospheric deposition and a measured response in vegetation has generally excluded the investigation of vegetation recovery from the impacts of atmospheric deposition. However, policy-makers require such evidence to assess whether policy decisions to reduce emissions will have a positive impact on habitats. Here we have shown that 40 years after the peak of SO
x emissions, decreases in SOx are related to significant changes in species richness and cover in Scottish Calcareous, Mestrophic, Nardus and Wet grasslands. Using a survey of vegetation plots across Scotland, first carried out between 1958 and 1987 and resurveyed between 2012 and 2014, we test whether temporal changes in species richness and cover of bryophytes, Cyperaceae, forbs, Poaceae, and Juncaceae can be explained by changes in sulphur and nitrogen deposition, climate and/or grazing intensity, and whether these patterns differ between six grassland habitats: Acid, Calcareous, Lolium, Nardus, Mesotrophic and Wet grasslands. The results indicate that Calcareous, Mesotrophic, Nardus and Wet grasslands in Scotland are starting to recover from the UK peak of SOx deposition in the 1970's. A decline in the cover of grasses, an increase in cover of bryophytes and forbs and the development of a more diverse sward (a reversal of the impacts of increased SOx ) was related to decreased SOx deposition. However there was no evidence of a recovery from SOx deposition in the Acid or Lolium grasslands. Despite a decline in NOx deposition between the two surveys we found no evidence of a reversal of the impacts of increased N deposition. The climate also changed significantly between the two surveys, becoming warmer and wetter. This change in climate was related to significant changes in both the cover and species richness of bryophytes, Cyperaceae, forbs, Poaceae and Juncaceae but the changes differed between habitats., (Copyright © 2018 Elsevier Ltd. All rights reserved.)- Published
- 2018
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14. Examining Africentric Cultural Values, Ethnic Identity, and Substance Use Abstinence in Low-Income, Early Adolescent, African American Girls.
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Sanchez D, Hamilton ER, Gilbert DJ, and Vandewater EA
- Abstract
An examination of cultural protective factors that foster substance use abstinence among low-income, early adolescent, African American girls may be helpful in understanding how to promote resilience and reduce negative health outcomes. This study examined the relations between Africentric cultural values, ethnic identity, and substance use abstinence among 196 low-income African American early adolescent girls (age 11-14 years). Results of logistic regressions revealed that Africentric values were negatively linked to cigarette and alcohol abstinence. Results also showed a significant positive interaction between Africentric cultural values and ethnic identity exploration that contributed to increased cigarette and alcohol abstinence. Implications for research and practice with African American early adolescent girls are discussed., Competing Interests: Declaration of Conflicting Interests The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
- Published
- 2018
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15. Clinical symptoms, signs and tests for identification of impending and current water-loss dehydration in older people.
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Hooper L, Abdelhamid A, Attreed NJ, Campbell WW, Channell AM, Chassagne P, Culp KR, Fletcher SJ, Fortes MB, Fuller N, Gaspar PM, Gilbert DJ, Heathcote AC, Kafri MW, Kajii F, Lindner G, Mack GW, Mentes JC, Merlani P, Needham RA, Olde Rikkert MG, Perren A, Powers J, Ranson SC, Ritz P, Rowat AM, Sjöstrand F, Smith AC, Stookey JJ, Stotts NA, Thomas DR, Vivanti A, Wakefield BJ, Waldréus N, Walsh NP, Ward S, Potter JF, and Hunter P
- Subjects
- Aged, Dehydration blood, Electric Impedance, Female, Humans, Male, Mouth Diseases diagnosis, Osmolar Concentration, Sensitivity and Specificity, Skin Physiological Phenomena, Symptom Assessment methods, Urine, Dehydration diagnosis, Drinking Water administration & dosage
- Abstract
Background: There is evidence that water-loss dehydration is common in older people and associated with many causes of morbidity and mortality. However, it is unclear what clinical symptoms, signs and tests may be used to identify early dehydration in older people, so that support can be mobilised to improve hydration before health and well-being are compromised., Objectives: To determine the diagnostic accuracy of state (one time), minimally invasive clinical symptoms, signs and tests to be used as screening tests for detecting water-loss dehydration in older people by systematically reviewing studies that have measured a reference standard and at least one index test in people aged 65 years and over. Water-loss dehydration was defined primarily as including everyone with either impending or current water-loss dehydration (including all those with serum osmolality ≥ 295 mOsm/kg as being dehydrated)., Search Methods: Structured search strategies were developed for MEDLINE (OvidSP), EMBASE (OvidSP), CINAHL, LILACS, DARE and HTA databases (The Cochrane Library), and the International Clinical Trials Registry Platform (ICTRP). Reference lists of included studies and identified relevant reviews were checked. Authors of included studies were contacted for details of further studies., Selection Criteria: Titles and abstracts were scanned and all potentially relevant studies obtained in full text. Inclusion of full text studies was assessed independently in duplicate, and disagreements resolved by a third author. We wrote to authors of all studies that appeared to have collected data on at least one reference standard and at least one index test, and in at least 10 people aged ≥ 65 years, even where no comparative analysis has been published, requesting original dataset so we could create 2 x 2 tables., Data Collection and Analysis: Diagnostic accuracy of each test was assessed against the best available reference standard for water-loss dehydration (serum or plasma osmolality cut-off ≥ 295 mOsm/kg, serum osmolarity or weight change) within each study. For each index test study data were presented in forest plots of sensitivity and specificity. The primary target condition was water-loss dehydration (including either impending or current water-loss dehydration). Secondary target conditions were intended as current (> 300 mOsm/kg) and impending (295 to 300 mOsm/kg) water-loss dehydration, but restricted to current dehydration in the final review.We conducted bivariate random-effects meta-analyses (Stata/IC, StataCorp) for index tests where there were at least four studies and study datasets could be pooled to construct sensitivity and specificity summary estimates. We assigned the same approach for index tests with continuous outcome data for each of three pre-specified cut-off points investigated.Pre-set minimum sensitivity of a useful test was 60%, minimum specificity 75%. As pre-specifying three cut-offs for each continuous test may have led to missing a cut-off with useful sensitivity and specificity, we conducted post-hoc exploratory analyses to create receiver operating characteristic (ROC) curves where there appeared some possibility of a useful cut-off missed by the original three. These analyses enabled assessment of which tests may be worth assessing in further research. A further exploratory analysis assessed the value of combining the best two index tests where each had some individual predictive ability., Main Results: There were few published studies of the diagnostic accuracy of state (one time), minimally invasive clinical symptoms, signs or tests to be used as screening tests for detecting water-loss dehydration in older people. Therefore, to complete this review we sought, analysed and included raw datasets that included a reference standard and an index test in people aged ≥ 65 years.We included three studies with published diagnostic accuracy data and a further 21 studies provided datasets that we analysed. We assessed 67 tests (at three cut-offs for each continuous outcome) for diagnostic accuracy of water-loss dehydration (primary target condition) and of current dehydration (secondary target condition).Only three tests showed any ability to diagnose water-loss dehydration (including both impending and current water-loss dehydration) as stand-alone tests: expressing fatigue (sensitivity 0.71 (95% CI 0.29 to 0.96), specificity 0.75 (95% CI 0.63 to 0.85), in one study with 71 participants, but two additional studies had lower sensitivity); missing drinks between meals (sensitivity 1.00 (95% CI 0.59 to 1.00), specificity 0.77 (95% CI 0.64 to 0.86), in one study with 71 participants) and BIA resistance at 50 kHz (sensitivities 1.00 (95% CI 0.48 to 1.00) and 0.71 (95% CI 0.44 to 0.90) and specificities of 1.00 (95% CI 0.69 to 1.00) and 0.80 (95% CI 0.28 to 0.99) in 15 and 22 people respectively for two studies, but with sensitivities of 0.54 (95% CI 0.25 to 0.81) and 0.69 (95% CI 0.56 to 0.79) and specificities of 0.50 (95% CI 0.16 to 0.84) and 0.19 (95% CI 0.17 to 0.21) in 21 and 1947 people respectively in two other studies). In post-hoc ROC plots drinks intake, urine osmolality and axillial moisture also showed limited diagnostic accuracy. No test was consistently useful in more than one study.Combining two tests so that an individual both missed some drinks between meals and expressed fatigue was sensitive at 0.71 (95% CI 0.29 to 0.96) and specific at 0.92 (95% CI 0.83 to 0.97).There was sufficient evidence to suggest that several stand-alone tests often used to assess dehydration in older people (including fluid intake, urine specific gravity, urine colour, urine volume, heart rate, dry mouth, feeling thirsty and BIA assessment of intracellular water or extracellular water) are not useful, and should not be relied on individually as ways of assessing presence or absence of dehydration in older people.No tests were found consistently useful in diagnosing current water-loss dehydration., Authors' Conclusions: There is limited evidence of the diagnostic utility of any individual clinical symptom, sign or test or combination of tests to indicate water-loss dehydration in older people. Individual tests should not be used in this population to indicate dehydration; they miss a high proportion of people with dehydration, and wrongly label those who are adequately hydrated.Promising tests identified by this review need to be further assessed, as do new methods in development. Combining several tests may improve diagnostic accuracy.
- Published
- 2015
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16. Advancing the Africentric paradigm shift discourse: building toward evidence-based Africentric interventions in social work practice with African Americans.
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Gilbert DJ, Harvey AR, and Belgrave FZ
- Subjects
- Culture, HIV Infections prevention & control, Humans, Psychology, Substance-Related Disorders, Black or African American, Evidence-Based Practice, Social Work methods
- Abstract
For over a decade, a number of social work scholars have advocated for an Africentric paradigm shift in social work practice with African Americans; yet the paradigm shift has been slow in coming with respect to infusing Africentric theory and interventions into social work practice, education, and research. Interventions that infuse Africentric values (such as interdependence, collectivism, transformation, and spirituality) have been shown to create significant change across a number of areas important to social work practice with African Americans. However, a barrier to the full integration of Africentric models into social work practice is that Africentric programs lack cohesive documentation and replication and, thus, have limited potential to be established as evidence-based practices. The authors present an overview of various Africentric interventions, including their program components and methods of evaluation, with the aim of establishing guideposts or next steps in developing a discourse on Africentric interventions that are promising best practices or are emerging as evidence-based practices. The authors conclude with implications for social work practice, education, and research and a call for Africentric scholars to engage in increased discussion, dissemination of manualized treatments, and collaborative research to build the evidence-based Africentric knowledge base and foster replication of studies.
- Published
- 2009
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17. Incorporating photodynamic therapy into a medical and cosmetic dermatology practice.
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Gilbert DJ
- Subjects
- Humans, Skin Diseases drug therapy, Dermatology, Office Management, Photochemotherapy
- Abstract
Effective in the treatment of a growing variety of skin conditions, photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA) is rising rapidly in popularity among both practitioners and patients, despite the fact that many applications are not yet cleared by the Food and Drug Administration. Because of its versatility, safety, efficacy, cosmetic benefits, and potential financial advantages, ALA-PDT may soon be a mainstay in many clinical settings. This article provides an overview of this easy-to-use treatment modality and a guide to implementing ALA-PDT into practice, including pretreatment and post-treatment protocols and guidelines for managing side effects.
- Published
- 2007
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18. HIV prevention targeting African American women: theory, objectives, and outcomes from an African-Centered Behavior Change Perspective.
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Gilbert DJ and Goddard L
- Subjects
- Culture, Female, Humans, Models, Psychological, Sexual Behavior ethnology, United States ethnology, Women's Health, Black or African American, HIV Infections ethnology, HIV Infections prevention & control, Health Education methods, Health Promotion methods, Risk Reduction Behavior
- Published
- 2007
- Full Text
- View/download PDF
19. The use of photodynamic therapy in dermatology: results of a consensus conference.
- Author
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Nestor MS, Gold MH, Kauvar AN, Taub AF, Geronemus RG, Ritvo EC, Goldman MP, Gilbert DJ, Richey DF, Alster TS, Anderson RR, Bank DE, Carruthers A, Carruthers J, Goldberg DJ, Hanke CW, Lowe NJ, Pariser DM, Rigel DS, Robins P, Spencer JM, and Zelickson BD
- Subjects
- Clinical Trials as Topic, Humans, Aminolevulinic Acid therapeutic use, Carcinoma, Basal Cell drug therapy, Keratosis drug therapy, Photochemotherapy methods, Photosensitizing Agents therapeutic use, Skin Neoplasms drug therapy
- Abstract
Photodynamic therapy (PDT) has significant promise in improving outcomes of patients with a variety of cutaneous conditions. A group of experts met to review the principles, indications, and clinical benefits of PDT with 5-aminolevulinic acid (ALA). They also reviewed PDT with methyl aminolevulinate. The experts established consensus statements for pretreatment, posttreatment, ALA contact time, light sources, and numbers of sessions associated with ALA PDT for actinic keratosis and superficial basal cell carcinoma, photorejuvenation and cosmetic enhancement, acne, sebaceous skin, rosacea, and rhinophyma. They based consensus recommendations on their clinical experience and the medical literature. They also suggested future applications of ALA PDT. Experts concluded that ALA PDT is a safe and effective modality for the treatment of conditions commonly encountered in dermatology. Since downtime is minimal, the technique is suitable for patients of all ages and lifestyles. Appropriate light sources are available in many dermatology offices. The expanding clinical and financial benefits of PDT justify the purchase of an appropriate light source.
- Published
- 2006
20. Treatment of actinic keratoses with sequential combination of 5-fluorouracil and photodynamic therapy.
- Author
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Gilbert DJ
- Subjects
- Adult, Aged, Aged, 80 and over, Female, Humans, Male, Middle Aged, Aminolevulinic Acid administration & dosage, Facial Dermatoses drug therapy, Fluorouracil administration & dosage, Keratosis drug therapy, Photochemotherapy
- Abstract
Actinic keratoses (AKs) are traditionally treated with cryotherapy, curettage, and 5-fluorouracil (5-FU, Efudex, ICN Pharmaceuticals, Inc.), all of which are associated with adverse effects. Although photodynamic therapy (PDT) with topical 5-aminolevulinic acid (ALA) offers a treatment alternative, current protocols require 14 to 18 hours incubation with ALA and patients experience pain during light treatment. Fifteen patients with multiple and diffuse facial AKs applied 5-FU nightly for 5 days and underwent PDT with ALA (Levulan Kerastick, Dusa Pharmaceuticals, Inc.) on the sixth day. ALA was applied to their entire faces and remained in contact with the skin for 30 to 45 minutes under low-intensity visible light. After removing ALA, faces received a single pass of 560- to 1200-nm intense pulsed light (VascuLight or Lumenis One, Lumenis). At 1 month and at 1 year post-treatment, 90% of treated AKs had resolved in all but one patient. Erythema resolved 7 to 10 days after treatment. Patients with multiple diffuse AKs may benefit from the application of 5-FU for 5 days followed by ALA-PDT with intense pulsed light activation.
- Published
- 2005
21. The interferon-inducible p200 family of proteins: a perspective on their roles in cell cycle regulation and differentiation.
- Author
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Asefa B, Klarmann KD, Copeland NG, Gilbert DJ, Jenkins NA, and Keller JR
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- Cell Cycle Proteins genetics, Cell Cycle Proteins physiology, Cell Differentiation, Humans, Immune System Diseases etiology, Neoplasms etiology, Nuclear Proteins genetics, Nuclear Proteins physiology
- Abstract
The interferon-inducible p200 (IFI-200) family of proteins is among the numerous gene products induced by interferons (IFNs), which are important regulators of cell growth, immunomodulation and host resistance to tumors and viral infections. The members of this family of proteins are highly homologous to one another and consist of five murine proteins including p202, p203, p204 and p205 as well as three human homologues; IFI-16, myeloid cell nuclear differentiation antigen (MNDA) and absent in melanoma (AIM) 2. They possess at least one copy of a conserved 200 amino-acid motif which exists in two types; the a and b domains. Most of the IFI-200 proteins also possess a domain in apoptosis and interferon response (DAPIN)/PYRIN domain, which is a conserved motif associated with protein-protein interactions in the regulation of apoptotic and inflammatory signaling pathways. The p200 proteins have been implicated in cell cycle regulation and differentiation based on their ability to interact with and modulate the activities of multiple transcriptional factors such as Rb and p53, and there are significant findings that link mutations in their genetic loci to the incidence of cancer. Here, we describe the structure and biological activities of these proteins, and discuss recent studies that describe their relevant roles in processes regulating cell proliferation and differentiation.
- Published
- 2004
- Full Text
- View/download PDF
22. Identification and characterization of a new pair of immunoglobulin-like receptors LMIR1 and 2 derived from murine bone marrow-derived mast cells.
- Author
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Kumagai H, Oki T, Tamitsu K, Feng SZ, Ono M, Nakajima H, Bao YC, Kawakami Y, Nagayoshi K, Copeland NG, Gilbert DJ, Jenkins NA, Kawakami T, and Kitamura T
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cell Line, Chromosome Mapping, Cloning, Molecular, Hematopoietic Stem Cells physiology, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases, Phosphoric Monoester Hydrolases metabolism, Phosphorylation, Protein Structure, Tertiary, Protein Tyrosine Phosphatases metabolism, RNA, Messenger biosynthesis, Receptors, Immunologic chemistry, Tyrosine metabolism, Mast Cells immunology, Receptors, Immunologic genetics, Receptors, Immunologic metabolism
- Abstract
We have identified and characterized two mouse cDNAs in a mouse antigen-stimulated bone marrow-derived mast cell cDNA library, both of which encode type I transmembrane proteins. The genes were closely mapped in the distal region of mouse chromosome 11 and expressed not only in mast cells but also widely in leukocytes. The extracellular domains of their encoded proteins contain a single variable immunoglobulin (Ig) motif sharing about 90% identity with amino acids, showing that they comprise a pair of molecules and belong to the Ig superfamily. We named these molecules leukocyte mono-Ig-like receptor1 and 2 (LMIR1 and 2). The intracellular domain of LMIR1 contains several immunoreceptor tyrosine-based inhibition motifs (ITIMs). When cross-linked, the intracellular domain was tyrosine phosphorylated and capable of recruiting tyrosine phosphatases, SHP-1 and SHP-2 and inositol polyphosphate 5-phosphatase, SHIP. LMIR2, on the other hand, contains a short cytoplasmic tail and a characteristic transmembrane domain carrying two positively charged amino acids associated with three kinds of immunoreceptor tyrosine-based activation motif (ITAM)-bearing molecules, DAP10, DAP12, and FcRgamma. These findings suggest that a new pair of ITIM/ITAM-bearing receptors, LMIR1 and 2, regulate mast cell-mediated inflammatory responses through yet to be defined ligand(s).
- Published
- 2003
- Full Text
- View/download PDF
23. Identification of beta-carotene 15, 15'-monooxygenase as a peroxisome proliferator-activated receptor target gene.
- Author
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Boulanger A, McLemore P, Copeland NG, Gilbert DJ, Jenkins NA, Yu SS, Gentleman S, and Redmond TM
- Subjects
- Animals, Binding Sites, Caco-2 Cells, Cell Line, Cloning, Molecular, Humans, Mice, Models, Genetic, Receptors, Cytoplasmic and Nuclear agonists, Response Elements, Transcription Factors agonists, beta-Carotene 15,15'-Monooxygenase, Oxygenases genetics, Promoter Regions, Genetic, Receptors, Cytoplasmic and Nuclear metabolism, Transcription Factors metabolism, Transcriptional Activation
- Abstract
Beta-carotene 15,15'-monooxygenase (BCM) catalyzes the first step of vitamin A biosynthesis from provitamin A carotenoids. We wished to determine the factors underlying the transcriptional regulation of this gene. After cloning of the 40 kilobase pair (kbp) mouse Bcm gene and determination of its genomic organization, analysis of the 2 kb 5'-flanking region showed several putative transcription factor binding sites including TATA box, a peroxisome proliferator response element (PPRE), AP2, and bHLH. The 2 kb fragment drove specific luciferase gene expression in vitro only in cell lines that express BCM (TC7, PF11, and monkey retinal pigment epithelium). Nucleotides -41 to +163, and -60 to +163 drove basal and specific Bcm transcriptional activity, respectively. Site-directed mutagenesis and gel shift experiments demonstrate that PPRE was essential for Bcm promoter specificity and that the peroxisome proliferator activated receptor (PPAR) gamma (PPARgamma) specifically binds to this element. Furthermore, cotransfection experiments and pharmacological treatments in vitro, using the specific PPARgamma agonists LY17883 and ciglitazone, demonstrate that the PPRE element confers peroxisome proliferator responsiveness via the PPARgamma and retinoid X receptor-alpha heterodimer. Treatment of mice with the PPARalpha/gamma agonist WY14643 increases BCM protein expression in liver. Thus PPAR is a key transcription factor for the transcriptional regulation of the Bcm gene, suggesting a broader function for PPARs in the regulation of carotenoid metabolism metabolism that is consistent with their established role in neutral lipid metabolism and transport.
- Published
- 2003
- Full Text
- View/download PDF
24. Common sites of retroviral integration in mouse hematopoietic tumors identified by high-throughput, single nucleotide polymorphism-based mapping and bacterial artificial chromosome hybridization.
- Author
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Shen H, Suzuki T, Munroe DJ, Stewart C, Rasmussen L, Gilbert DJ, Jenkins NA, and Copeland NG
- Subjects
- Animals, Chromosome Mapping, Mice, Mutagenesis, Insertional, Retroviridae genetics, Chromosomes, Artificial, Bacterial, Hematologic Neoplasms virology, Polymorphism, Single Nucleotide, Retroviridae physiology, Virus Integration
- Abstract
Retroviral insertional mutagenesis in mouse hematopoietic tumors provides a powerful cancer gene discovery tool. Here, we describe a high-throughput, single nucleotide polymorphism (SNP)-based method, for mapping retroviral integration sites cloned from mouse tumors, and a bacterial artificial chromosome (BAC) hybridization method, for localizing these retroviral integration sites to common sites of retroviral integration (CISs). Several new CISs were identified, including one CIS that mapped near Notch1, a gene that has been causally associated with human T-cell tumors. This mapping method is applicable to many different species, including ones where few genetic markers and little genomic sequence information are available. It can also be used to map endogenous proviruses.
- Published
- 2003
- Full Text
- View/download PDF
25. The dual specificity JKAP specifically activates the c-Jun N-terminal kinase pathway.
- Author
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Chen AJ, Zhou G, Juan T, Colicos SM, Cannon JP, Cabriera-Hansen M, Meyer CF, Jurecic R, Copeland NG, Gilbert DJ, Jenkins NA, Fletcher F, Tan TH, and Belmont JW
- Subjects
- Amino Acid Sequence, Animals, Cell Line, Crosses, Genetic, DNA, Complementary metabolism, Dose-Response Relationship, Drug, Enzyme Activation, Hematopoietic Stem Cells metabolism, Humans, In Situ Hybridization, MAP Kinase Kinase 7, MAP Kinase Signaling System, Mice, Mice, Inbred C57BL, Mitogen-Activated Protein Kinase 1 metabolism, Mitogen-Activated Protein Kinases, Models, Genetic, Molecular Sequence Data, Mutation, Plasmids metabolism, Precipitin Tests, Protein Binding, RNA, Messenger metabolism, Sequence Homology, Amino Acid, Signal Transduction, Substrate Specificity, Tissue Distribution, Transfection, Transforming Growth Factor beta pharmacology, Ultraviolet Rays, p38 Mitogen-Activated Protein Kinases, JNK Mitogen-Activated Protein Kinases, MAP Kinase Kinase 4, Mitogen-Activated Protein Kinase Kinases metabolism, Phosphoprotein Phosphatases metabolism, Phosphoprotein Phosphatases physiology, Tumor Necrosis Factor-alpha metabolism
- Abstract
The involvement of dual specificity phosphatases (DSPs) in the mitogen-activated protein kinase (MAPK) signaling has been mostly limited to the inactivation of MAPKs by the direct dephosphorylation of the TXY motif within their activation loop. We report the cloning and characterization of a murine DSP, called JNK pathway-associated phosphatase (JKAP), which lacks the regulatory region present in most other MAP kinase phosphatases (MKPs) and is preferentially expressed in murine Lin(-)Sca-1(+) stem cells. Overexpression of JKAP in human embryonic kidney 293T cells specifically activated c-Jun N-terminal kinase (JNK) but not p38 and extracellular signal-regulated kinase 2. Overexpression of a mutant JKAP, JKAP-C88S, blocked tumor necrosis factor-alpha-induced JNK activation. Targeted gene disruption in murine embryonic stem cells abolished JNK activation by tumor necrosis factor-alpha and transforming growth factor-beta, but not by ultraviolet-C irradiation, indicating that JKAP is necessary for optimal JNK activation. JKAP associated with JNK and MKK7, but not SEK1, in vivo. However, JKAP did not interact with JNK in vitro, suggesting that JKAP exerts its effect on JNK in an indirect manner. Taken together, these studies identify a positive regulator for the JNK pathway and suggest a novel role for DSP in mitogen-activated protein kinase regulation.
- Published
- 2002
- Full Text
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26. Murine NFX.1: isolation and characterization of its messenger RNA, mapping of its chromosomal location and assessment of its developmental expression.
- Author
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Arlotta P, Miyazaki D, Copeland NG, Gilbert DJ, Jenkins NA, and Ono SJ
- Subjects
- Aging immunology, Amino Acid Sequence, Animals, Cell Culture Techniques, DNA, Complementary genetics, DNA-Binding Proteins chemistry, Female, Genes, MHC Class II immunology, Humans, Male, Mice, Inbred C57BL, Molecular Sequence Data, RNA, Messenger genetics, Repressor Proteins, Species Specificity, Transcription Factors, Chromosome Mapping, DNA-Binding Proteins genetics, Gene Expression Regulation, Developmental immunology, Mice immunology
- Abstract
We have previously isolated (by expression cloning) a human cDNA, termed NFX.1, encoding a nucleic acid-binding protein that interacts with the conserved X1 box cis-element first discovered in class II major histocompatibility complex (MHC) genes. Functional studies involving expression of NFX.1 and assessment of expression from class II reporter constructs and endogenous class II MHC genes indicated that the factor could repress transcription of class II MHC genes. Subsequent studies have extended the biological significance of the factor, indicating that it plays an important role in neuronal development. Indeed, the reiterated RING finger motifs in the central domain of the polypeptide strongly suggest that NF-XI is a probable E3 ubiquitin protein ligase, indicating that the protein may have multiple activities. Here we report the cloning of the mouse homologue of the human NfX.1 cDNA: m-Nfx.1. Comparison of the deduced primary sequence of mouse and human NFX.1 proteins shows very high homology and confirms that m-NFX.1 contains the conserved cysteine-rich DNA-binding motif first described in human NFX.1 (95% homology). Expression of MHC class II genes is substantially reduced following expression of m-NFX.1, which confirms that we have isolated the functional murine homologue of human NfX.1 cDNA. Further evidence comes from the mapping of m-Nfx.1 gene to the proximal region of mouse chromosome 4, a region syntenic to the location of human Nfx.1 (short arm of chromosome 9). Expression profiling shows that m-NFX.1 is expressed ubiquitously in both adult tissues and during development, supporting the hypothesis that it may have yet-undescribed roles in distinct biological processes.
- Published
- 2002
- Full Text
- View/download PDF
27. Comparative genetics and evolution of annexin A13 as the founder gene of vertebrate annexins.
- Author
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Iglesias JM, Morgan RO, Jenkins NA, Copeland NG, Gilbert DJ, and Fernandez MP
- Subjects
- Alternative Splicing, Amino Acid Sequence, Animals, Base Sequence, Cell Line, Crosses, Genetic, DNA, Complementary genetics, Female, Founder Effect, Genes, Regulator, Humans, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Muridae, Rats, Sequence Homology, Amino Acid, Species Specificity, Annexins genetics, Evolution, Molecular, Vertebrates genetics
- Abstract
Annexin A13 (ANXA13) is believed to be the original founder gene of the 12-member vertebrate annexin A family, and it has acquired an intestine-specific expression associated with a highly differentiated intracellular transport function. Molecular characterization of this subfamily in a range of vertebrate species was undertaken to assess coding region conservation, gene organization, chromosomal linkage, and phylogenetic relationships relevant to its progenitor role in the structure-function evolution of the annexin gene superfamily. Protein diagnostic features peculiar to this subfamily include an alternate isoform containing a KGD motif, an elevated basic amino acid content with polyhistidine expansion in the 5'-translated region, and the conservation of 15% core tetrad residues specific to annexin A13 members. The 12 coding exons comprising the 58-kb human ANXA13 gene were deduced from BAC clone sequencing, whereas internal repetitive elements and neighboring genes in chromosome 8q24.12 were identified by contig analysis of the draft sequence from the human genome project. A unique exon splicing pattern in the annexin A13 gene was corroborated by coanalysis of mouse, rat, zebrafish, and pufferfish genomic DNA and determined to be the most distinct of all vertebrate annexins. The putative promoter region was identified by phylogenetic footprinting of potential binding sites for intestine-specific transcription factors. Mouse annexin A13 cDNA was used to map the gene to an orthologous linkage group in mouse chromosome 15 (between Sdc2 and Myc by backcross analysis), and the zebrafish cDNA permitted its localization to linkage group 24. Comparative analysis of annexin A13 from nine species traced this gene's speciation history and assessed coding region variation, whereas phylogenetic analysis showed it to be the deepest-branching vertebrate annexin, and computational analysis estimated the gene age and divergence rate. The unique, conserved aspects of annexin A13 primary structure, gene organization, and genetic maps identify it as the probable common ancestor of all vertebrate annexins, beginning with the sequential duplication to annexins A7 and A11 approximately 700 MYA, before the emergence of chordates.
- Published
- 2002
- Full Text
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28. Differential expression of the seven-pass transmembrane cadherin genes Celsr1-3 and distribution of the Celsr2 protein during mouse development.
- Author
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Shima Y, Copeland NG, Gilbert DJ, Jenkins NA, Chisaka O, Takeichi M, and Uemura T
- Subjects
- Animals, Blotting, Western, Cell Line, Cells, Cultured, Central Nervous System embryology, Chromosome Mapping, Cloning, Molecular, Crosses, Genetic, Ear, Inner metabolism, Hippocampus metabolism, Humans, Immunoblotting, In Situ Hybridization, Mice, Models, Genetic, Neurons metabolism, Purkinje Cells metabolism, RNA metabolism, RNA, Messenger metabolism, Time Factors, Tissue Distribution, Transfection, Cadherins biosynthesis, Cadherins metabolism, Fetal Proteins, Receptors, Cell Surface biosynthesis, Receptors, G-Protein-Coupled
- Abstract
Drosophila Flamingo (Fmi) is an evolutionally conserved seven-pass transmembrane receptor of the cadherin superfamily. Fmi plays multiple roles in patterning neuronal processes and epithelial planar cell polarity. To explore the in vivo roles of Fmi homologs in mammals, we previously cloned one of the mouse homologs, mouse flamingo1/Celsr2. Here, we report the results of our study of its embryonic and postnatal expression patterns together with those of two other paralogs, Celsr1 and Celsr3. Celsr1-3 expression was initiated broadly in the nervous system at early developmental stages, and each paralog showed characteristic expression patterns in the developing CNS. These genes were also expressed in several other organs, including the cochlea, where hair cells develop planar polarity, the kidney, and the whisker. The Celsr2 protein was distributed at intercellular boundaries in the whisker and on processes of neuronal cells such as hippocampal pyramidal cells, Purkinje cells, and olfactory neurons. Celsr2 is mapped to a distal region of the mouse chromosome 3. We discussed possible functions of seven-pass transmembrane cadherins in mouse development.
- Published
- 2002
- Full Text
- View/download PDF
29. Significant differences between mouse and human trophinins are revealed by their expression patterns and targeted disruption of mouse trophinin gene.
- Author
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Nadano D, Sugihara K, Paria BC, Saburi S, Copeland NG, Gilbert DJ, Jenkins NA, Nakayama J, and Fukuda MN
- Subjects
- Amino Acid Sequence, Animals, Blastocyst physiology, Blotting, Southern, Blotting, Western, Cell Adhesion Molecules biosynthesis, Chromosome Mapping, Cloning, Molecular, Female, Gene Expression Regulation, Developmental, Genome, Genotype, Humans, Immunohistochemistry, Inbreeding, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Reverse Transcriptase Polymerase Chain Reaction, Species Specificity, X Chromosome genetics, Cell Adhesion Molecules genetics, Cell Adhesion Molecules physiology
- Abstract
Trophinin has been identified as a membrane protein mediating apical cell adhesion between two human cell lines: trophoblastic HT-H cells, and endometrial epithelial SNG-M cells. Expression patterns of trophinin in humans suggested its involvement in embryo implantation and early placental development. The mouse trophinin gene maps to the distal part of the X chromosome and corresponds to human chromosome Xp11.21-22, the locus where the human trophinin gene maps. Western blot analysis indicates that the molecular weight of mouse trophinin is 110 kDa, which is consistent with the calculated value of 107 kDa. Positive signals for trophinin proteins were detected in preimplantation mouse embryos at the morula and blastocyst stages. Implanting blastocysts do not show detectable levels of trophinin protein, demonstrating that trophinin is not involved in blastocyst adhesion to the uterus in the mouse. Mouse embryo strongly expressed trophinin in the epiblast 1 day after implantation. Trophinin protein was not found in the mouse uteri and placenta after 5.5 days postcoitus (dpc). Targeted disruption of the trophinin gene in the mouse showed a partial embryonic lethality in a 129/SvJ background, but the cause of this lethality remains undetermined. The present study indicates significant differences between mouse and human trophinins in their expression patterns, and it suggests that trophinin is not involved in embryo implantation and placental development in the mouse.
- Published
- 2002
- Full Text
- View/download PDF
30. Evolution of the regulators of G-protein signaling multigene family in mouse and human.
- Author
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Sierra DA, Gilbert DJ, Householder D, Grishin NV, Yu K, Ukidwe P, Barker SA, He W, Wensel TG, Otero G, Brown G, Copeland NG, Jenkins NA, and Wilkie TM
- Subjects
- Animals, Evolution, Molecular, Female, Humans, Male, Mice, Mice, Inbred C57BL, Phylogeny, Signal Transduction, Chromosome Mapping, Multigene Family, RGS Proteins genetics
- Abstract
The regulators of G-protein signaling (RGS) proteins are important regulatory and structural components of G-protein coupled receptor complexes. RGS proteins are GTPase activating proteins (GAPs) of Gi-and Gq-class Galpha proteins, and thereby accelerate signaling kinetics and termination. Here, we mapped the chromosomal positions of all 21 Rgs genes in mouse, and determined human RGS gene structures using genomic sequence from partially assembled bacterial artificial chromosomes (BACs) and Celera fragments. In mice and humans, 18 of 21 RGS genes are either tandemly duplicated or tightly linked to genes encoding other components of G-protein signaling pathways, including Galpha, Ggamma, receptors (GPCR), and receptor kinases (GPRK). A phylogenetic tree revealed seven RGS gene subfamilies in the yeast and metazoan genomes that have been sequenced. We propose that similar systematic analyses of all multigene families from human and other mammalian genomes will help complete the assembly and annotation of the human genome sequence.
- Published
- 2002
- Full Text
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31. Cloning, expression, and chromosomal localization of the mouse gene (Scgb3a1, alias Ugrp2) that encodes a member of the novel uteroglobin-related protein gene family.
- Author
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Niimi T, Copeland NG, Gilbert DJ, Jenkins NA, Srisodsai A, Zimonjic DB, Keck-Waggoner CL, Popescu NC, and Kimura S
- Subjects
- Alternative Splicing, Amino Acid Sequence, Animals, Base Sequence, Chromosome Mapping, Chromosomes, Human, Pair 5 genetics, Cloning, Molecular, DNA, Complementary genetics, Female, Gene Expression, Humans, In Situ Hybridization, Fluorescence, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Multigene Family, Muridae, RNA, Messenger genetics, RNA, Messenger metabolism, Species Specificity, Proteins genetics
- Abstract
The mouse UGRP gene family consists of two genes, Ugrp1 and Ugrp2. In this study, the genomic structure and expression patterns of Ugrp2 and its alternative spliced form were characterized. The authentic Ugrp2 gene has three exons and two introns, similar to the Ugrp1 gene, which produces a secreted protein. The Ugrp2 variant uses a sequence located between authentic exons 1 and 2, resulting in a cytoplasmic form due to a termination codon within the inserted sequence. Both mouse and human UGRP2 mRNAs are expressed in lung. In the case of human, the mRNA is expressed at the highest level in trachea, followed by salivary gland at a level similar to lung. Weak expression was also found in fetal lung and mammary gland. Ugrp2 was mapped by fluorescence in situ hybridization to mouse chromosome 11A5-B1 and human chromosome 5q35. These regions are known to be homologous. Interspecific mouse backcross mapping was also performed to obtain further detailed localization of mouse Ugrp1 and Ugrp2., (Copyright 2002 S. Karger AG, Basel)
- Published
- 2002
- Full Text
- View/download PDF
32. The murine cysteinyl leukotriene 2 (CysLT2) receptor. cDNA and genomic cloning, alternative splicing, and in vitro characterization.
- Author
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Hui Y, Yang G, Galczenski H, Figueroa DJ, Austin CP, Copeland NG, Gilbert DJ, Jenkins NA, and Funk CD
- Subjects
- 5' Untranslated Regions, Adrenal Glands metabolism, Amino Acid Sequence, Animals, Base Sequence, Binding, Competitive, Blotting, Northern, Calcium metabolism, Cell Line, Cells, Cultured, Chromosome Mapping, Cloning, Molecular, Crosses, Genetic, Dose-Response Relationship, Drug, Exons, Humans, In Situ Hybridization, Introns, Leukotriene C4 metabolism, Leukotriene D4 metabolism, Mice, Mice, Inbred C57BL, Models, Genetic, Molecular Sequence Data, Myocardium metabolism, Protein Structure, Tertiary, RNA, Messenger metabolism, Radioligand Assay, Recombinant Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Sequence Homology, Amino Acid, Spleen metabolism, Thymus Gland metabolism, Tissue Distribution, Transfection, Alternative Splicing, DNA, Complementary metabolism, Membrane Proteins, Receptors, Leukotriene chemistry, Receptors, Leukotriene genetics
- Abstract
Two classes of cysteinyl leukotriene receptor, CysLT(1) and CysLT(2), have been identified and pharmacologically characterized in human tissues. Although the CysLT(1) receptor mediates the proinflammatory effects of leukotrienes in human asthma, the physiological roles of CysLT(2) receptor are not defined, and a suitable mouse model would be useful in delineating function. We report here the molecular cloning and characterization of the mouse CysLT(2) receptor (mCysLT(2)R) from heart tissue. mCysLT(2)R cDNA encodes a protein of 309 amino acids, truncated at both ends compared with the human ortholog (hCysLT(2)R). The gene resides on the central region of mouse chromosome 14 and is composed of 6 exons with the entire coding region located in the last exon. Two 5'-untranslated region splice variants were identified with the short form lacking exon 3 as the predominant transcript. Although the overall expression of mCysLT(2)R is very low, the highest expression was detected in spleen, thymus, and adrenal gland by ribonuclease protection assay, and discrete sites of expression in heart were observed by in situ hybridization. Intracellular calcium mobilization in response to cysteinyl leukotriene administration was detected in human embryonic kidney 293T cells transfected with recombinant mCysLT(2)R with a rank order of potency leukotriene C(4)(LTC(4) ) = LTD(4)>>LTE(4). [(3)H]LTD(4) binding to membranes expressing mCysLT(2)R could be effectively competed by LTC(4) and LTD(4) and only partially inhibited by LTE(4) and BAYu9773. The identification of mCysLT(2)R will be useful for establishing CysLT(2)R-deficient mice and determining novel leukotriene functions.
- Published
- 2001
- Full Text
- View/download PDF
33. Molecular characterization of the mouse Tem1/endosialin gene regulated by cell density in vitro and expressed in normal tissues in vivo.
- Author
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Opavsky R, Haviernik P, Jurkovicova D, Garin MT, Copeland NG, Gilbert DJ, Jenkins NA, Bies J, Garfield S, Pastorekova S, Oue A, and Wolff L
- Subjects
- 3T3 Cells, Amino Acid Sequence, Animals, Antigens, CD, Antigens, Neoplasm, Base Sequence, Blotting, Northern, Blotting, Southern, Blotting, Western, Cell Division, Cell Line, Cells, Cultured, Chromosome Mapping, Chromosomes, Human, Pair 19, Corpus Luteum metabolism, Crosses, Genetic, DNA, Complementary metabolism, Endothelium, Vascular cytology, Female, Gene Library, Humans, Immunohistochemistry, In Situ Hybridization, Introns, Luciferases metabolism, Mice, Mice, Inbred C57BL, Microscopy, Confocal, Microscopy, Fluorescence, Models, Genetic, Molecular Sequence Data, Promoter Regions, Genetic, Protein Binding, Reverse Transcriptase Polymerase Chain Reaction, Tissue Distribution, Transcription, Genetic, Up-Regulation, Membrane Proteins biosynthesis, Membrane Proteins genetics, Neoplasm Proteins biosynthesis, Neoplasm Proteins genetics
- Abstract
Human tumor endothelial marker 1/endosialin (TEM1/endosialin) was recently identified as a novel tumor endothelial cell surface marker potentially involved in angiogenesis, although no specific function for this novel gene has been assigned so far. It was reported to be expressed in tumor endothelium but not in normal endothelium with the exception of perhaps the corpus luteum. Here we describe the cDNA and genomic sequences for the mouse Tem1/endosialin homolog, the identification and characterization of its promoter region, and an extensive characterization of its expression pattern in murine and human tissues and murine cell lines in vitro. The single copy gene that was mapped to chromosome 19 is intronless and encodes a 92-kDa protein that has 77.5% overall homology to the human protein. The remarkable findings are 1) this gene is ubiquitously expressed in normal human and mouse somatic tissues and during development, and 2) its expression at the mRNA level is density-dependent and up-regulated in serum-starved cells. In vitro, its expression is limited to cells of embryonic, endothelial, and preadipocyte origin, suggesting that the wide distribution of its expression in vivo is due to the presence of vascular endothelial cells in all the tissues. The ubiquitous expression in vivo is in contrast to previously reported expression limited to corpus luteum and highly angiogenic tissues such as tumors and wound tissue.
- Published
- 2001
- Full Text
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34. Retroviral integration at the Epi1 locus cooperates with Nf1 gene loss in the progression to acute myeloid leukemia.
- Author
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Blaydes SM, Kogan SC, Truong BT, Gilbert DJ, Jenkins NA, Copeland NG, Largaespada DA, and Brannan CI
- Subjects
- Acute Disease, Animals, Gene Deletion, Genes, Neurofibromatosis 1, Genes, myb, Genetic Predisposition to Disease, Leukemia, Experimental etiology, Leukemia, Experimental virology, Leukemia, Myeloid etiology, Leukemia, Myeloid virology, Mice, Retroviridae genetics, Virus Integration genetics, Leukemia, Experimental genetics, Leukemia, Myeloid genetics
- Abstract
Juvenile myelomonocytic leukemia (JMML) is a disease that occurs in young children and is associated with a high mortality rate. In most patients, JMML has a progressive course leading to death by virtue of infection, bleeding, or progression to acute myeloid leukemia (AML). As it is known that children with neurofibromatosis type 1 syndrome have a markedly increased risk of developing JMML, we have previously developed a mouse model of JMML through reconstitution of lethally irradiated mice with hematopoietic stem cells homozygous for a loss-of-function mutation in the Nf1 gene (D. L. Largaespada, C. I. Brannan, N. A. Jenkins, and N. G. Copeland, Nat. Genet. 12:137-143, 1996). In the course of these experiments, we found that all these genetically identical reconstituted mice developed a JMML-like disorder, but only a subset went on to develop more acute disease. This result strongly suggests that additional genetic lesions are responsible for disease progression to AML. Here, we describe the production of a unique tumor panel, created using the BXH-2 genetic background, for identification of these additional genetic lesions. Using this tumor panel, we have identified a locus, Epi1, which maps 30 to 40 kb downstream of the Myb gene and appears to be the most common site of somatic viral integration in BXH-2 mice. Our findings suggest that proviral integrations at Epi1 cooperate with loss of Nf1 to cause AML.
- Published
- 2001
- Full Text
- View/download PDF
35. Nuclear membrane protein LAP2beta mediates transcriptional repression alone and together with its binding partner GCL (germ-cell-less).
- Author
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Nili E, Cojocaru GS, Kalma Y, Ginsberg D, Copeland NG, Gilbert DJ, Jenkins NA, Berger R, Shaklai S, Amariglio N, Brok-Simoni F, Simon AJ, and Rechavi G
- Subjects
- Amino Acid Sequence, Animals, Carcinoma, Non-Small-Cell Lung metabolism, Carrier Proteins physiology, Chromosomes chemistry, DNA-Binding Proteins chemistry, Drosophila, E2F Transcription Factors, Humans, Insulinoma metabolism, Intercellular Signaling Peptides and Proteins, Lung Neoplasms, Macromolecular Substances, Membrane Proteins chemistry, Mice, Molecular Sequence Data, Nuclear Envelope chemistry, Nuclear Proteins genetics, Nuclear Proteins isolation & purification, Pancreas cytology, Pancreas metabolism, Protein Transport physiology, Saccharomyces cerevisiae, Sequence Homology, Transcription, Genetic physiology, Tumor Cells, Cultured metabolism, Cell Cycle Proteins, DNA-Binding Proteins metabolism, Drosophila Proteins, Membrane Proteins metabolism, Nuclear Envelope metabolism, Nuclear Proteins metabolism, Transcription Factors metabolism
- Abstract
LAP2beta is an integral membrane protein of the nuclear envelope involved in chromatin and nuclear architecture. Using the yeast two-hybrid system, we have cloned a novel LAP2beta-binding protein, mGCL, which contains a BTB/POZ domain and is the mouse homologue of the Drosophila germ-cell-less (GCL) protein. In Drosophila embryos, GCL was shown to be essential for germ cell formation and was localized to the nuclear envelope. Here, we show that, in mammalian cells, GCL is co-localized with LAP2beta to the nuclear envelope. Nuclear fractionation studies reveal that mGCL acts as a nuclear matrix component and not as an integral protein of the nuclear envelope. Recently, mGCL was found to interact with the DP3alpha component of the E2F transcription factor. This interaction reduced the transcriptional activity of the E2F-DP heterodimer, probably by anchoring the complex to the nuclear envelope. We demonstrate here that LAP2beta is also capable of reducing the transcriptional activity of the E2F-DP complex and that it is more potent than mGCL in doing so. Co-expression of both LAP2beta and mGCL with the E2F-DP complex resulted in a reduced transcriptional activity equal to that exerted by the pRb protein.
- Published
- 2001
- Full Text
- View/download PDF
36. The murine perilipin gene: the lipid droplet-associated perilipins derive from tissue-specific, mRNA splice variants and define a gene family of ancient origin.
- Author
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Lu X, Gruia-Gray J, Copeland NG, Gilbert DJ, Jenkins NA, Londos C, and Kimmel AR
- Subjects
- 5' Untranslated Regions genetics, Amino Acid Sequence, Animals, Base Sequence, Carrier Proteins, DNA Primers chemistry, DNA, Complementary isolation & purification, DNA, Complementary metabolism, Dictyostelium genetics, Drosophila genetics, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Perilipin-1, Polyadenylation, Polymerase Chain Reaction, Polymorphism, Restriction Fragment Length, Sequence Homology, Amino Acid, Alternative Splicing genetics, Phosphoproteins genetics, RNA, Messenger genetics
- Abstract
The Perilipins are a family of intracellular neutral lipid droplet storage proteins that are responsive to acute protein kinase A-mediated, hormonal stimulation. Perilipin (Peri) expression appears to be limited to adipocytes and steroidogenic cells, in which intracellular neutral lipid hydrolysis is regulated by protein kinase A. We have isolated cDNA sets and overlapping genomic fragments of the murine Peri locus and mapped chromosomal location, transcription start sites, polyadenylylation sites, and intron/exon junctions. Data confirm that the Perilipins are encoded by a single-copy gene, with alternative and tissue-specific, mRNA splicing and polyadenylylation yielding four different protein species. The Perilipin proteins have identical approximately 22-kDa amino termini with distinct carboxyl terminal sequences of varying lengths. These genomic and transcriptional maps of murine Perilipin are also essential for evaluating presumptive endogenous and targeted mutations within the locus. The N-terminal identity region of the Perilipins defines a sequence motif, which we term PAT, that is shared with the ADRP and TIP47 proteins; additionally, the PAT domain may represent a novel, conserved pattern for lipid storage droplet (LSD) proteins of vertebrates and invertebrates alike. Comparative genomics suggest the presence of related LSD genes in species as diverse as Drosophila and Dictyostelium.
- Published
- 2001
- Full Text
- View/download PDF
37. STK25 is a candidate gene for pseudopseudohypoparathyroidism.
- Author
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Davids MS, Crawford E, Weremowicz S, Morton CC, Copeland NG, Gilbert DJ, Jenkins NA, Phelan MC, Comb MJ, and Melnick MB
- Subjects
- Animals, Chromosome Deletion, Chromosome Mapping, Chromosomes, Human, Pair 2 genetics, Female, Genetic Predisposition to Disease genetics, Humans, In Situ Hybridization, Fluorescence, Intracellular Signaling Peptides and Proteins, MAP Kinase Kinase Kinases, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Muridae, Pseudopseudohypoparathyroidism enzymology, Protein Serine-Threonine Kinases genetics, Pseudopseudohypoparathyroidism genetics, Saccharomyces cerevisiae Proteins
- Abstract
We determined the chromosomal location of the mouse gene Stk25, encoding a member of the Ste20/PAK family of serine/threonine kinases, by interspecific backcross analysis. We mapped Stk25 to the central region of mouse chromosome 1 linked to Chrng (formerly Acrg) and En1. This central region of mouse chromosome 1 shares a region of homology with the long arm of human chromosome 2, suggesting that the human homologue of Stk25 would also map to 2q. We proved this prediction of syntenic homology correct by mapping human STK25 to 2q37. Deletion of the 2q37 region has been implicated in the expression of pseudopseudohypoparathyroidism (PPHP), a disease which shares features of the Albright hereditary osteodystrophy (AHO) phenotype. To investigate a pathogenetic relationship between STK25 and PPHP, we carried out fluorescence in situ hybridization (FISH) using an STK25 gene probe and chromosomes from PPHP patients characterized as having small deletions near the distal end of 2q. PPHP patient DNA showed no hybridization to STK25 genomic DNA, indicating that STK25 is contained within the deleted chromosomal region. This finding, in conjunction with previous studies demonstrating the role of Ste20/PAK kinases in heterotrimeric G protein signaling, suggests that STK25 is a positional candidate gene for PPHP.
- Published
- 2001
- Full Text
- View/download PDF
38. The paralemmin protein family: identification of paralemmin-2, an isoform differentially spliced to AKAP2/AKAP-KL, and of palmdelphin, a more distant cytosolic relative.
- Author
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Hu B, Copeland NG, Gilbert DJ, Jenkins NA, and Kilimann MW
- Subjects
- A Kinase Anchor Proteins, Alternative Splicing, Animals, Chickens, Chromosomes, Human, Pair 1 genetics, Chromosomes, Human, Pair 9 genetics, Cytosol metabolism, DNA, Complementary genetics, DNA, Complementary isolation & purification, Humans, Membrane Proteins metabolism, Mice, Molecular Sequence Data, Organ Specificity, Phosphoproteins, Physical Chromosome Mapping, Protein Biosynthesis, Protein Isoforms genetics, Protein Isoforms metabolism, Protein Structure, Tertiary, Proteins metabolism, RNA, Messenger analysis, RNA, Messenger biosynthesis, Rats, Recombinant Fusion Proteins chemistry, Recombinant Fusion Proteins genetics, Sequence Alignment, Sequence Homology, Amino Acid, Terminology as Topic, Adaptor Proteins, Signal Transducing, Carrier Proteins genetics, Membrane Proteins genetics, Multigene Family, Proteins genetics
- Abstract
Paralemmin is a protein implicated in plasma membrane dynamics. Here we describe the identification of two new paralemmin-related proteins. A partial paralemmin homolog, palmdelphin, is predominantly cytosolic, unlike paralemmin which is lipid-anchored to the plasma membrane through a C-terminal CaaX motif. We have mapped the mouse palmdelphin gene to distal chromosome 3 between Amy2 and Abcd3, in a region homologous to human chromosome 1p22-p21 where the human palmdelphin gene is located. We have also identified a second paralemmin isoform, paralemmin-2. It is expressed from a gene on human chromosome 9q31-q33 which ends only 33 kb upstream of the gene encoding the protein kinase A-binding protein,AKAP2/AKAP-KL. The closely adjacent paralemmin-2 and AKAP2 genes are functionally linked in a very unusual manner. Chimeric mRNAs are expressed, apparently by RNA readthrough and differential splicing, that encode natural fusion proteins in which either the N-terminal coiled-coil region or nearly the complete sequence of paralemmin-2 except its C-terminal CaaX motif is fused to AKAP2/AKAP-KL. The N-terminal coiled-coil region is conserved in paralemmin-1, paralemmin-2/AKAP2, palmdelphin and a fourth, uncharacterized gene, suggesting that it is a modular functional domain., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
- View/download PDF
39. Murine homolog of SALL1 is essential for ureteric bud invasion in kidney development.
- Author
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Nishinakamura R, Matsumoto Y, Nakao K, Nakamura K, Sato A, Copeland NG, Gilbert DJ, Jenkins NA, Scully S, Lacey DL, Katsuki M, Asashima M, and Yokota T
- Subjects
- Amino Acid Sequence, Animals, Cloning, Molecular, Crosses, Genetic, Down-Regulation, Genetic Markers genetics, Heterozygote, Humans, In Situ Hybridization, Mesoderm metabolism, Mice, Mice, Inbred C57BL, Models, Genetic, Molecular Sequence Data, Mutation, Phenotype, Polymerase Chain Reaction, Sequence Homology, Amino Acid, Gene Expression Regulation, Developmental, Kidney embryology, Transcription Factors genetics, Transcription Factors physiology, Ureter embryology
- Abstract
SALL1 is a mammalian homolog of the Drosophila region-specific homeotic gene spalt (sal); heterozygous mutations in SALL1 in humans lead to Townes-Brocks syndrome. We have isolated a mouse homolog of SALL1 (Sall1) and found that mice deficient in Sall1 die in the perinatal period and that kidney agenesis or severe dysgenesis are present. Sall1 is expressed in the metanephric mesenchyme surrounding ureteric bud; homozygous deletion of Sall1 results in an incomplete ureteric bud outgrowth, a failure of tubule formation in the mesenchyme and an apoptosis of the mesenchyme. This phenotype is likely to be primarily caused by the absence of the inductive signal from the ureter, as the Sall1-deficient mesenchyme is competent with respect to epithelial differentiation. Sall1 is therefore essential for ureteric bud invasion, the initial key step for metanephros development.
- Published
- 2001
- Full Text
- View/download PDF
40. The genes encoding E-selectin (SELE) and lymphotactin (SCYC1) lie on separate chicken chromosomes although they are closely linked in human and mouse.
- Author
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Morroll S, Goodchild M, Salmon N, Copeland NG, Gilbert DJ, Jenkins NA, Bumstead N, and Boyd Y
- Subjects
- Animals, Chromosome Mapping, Conserved Sequence, Evolution, Molecular, Expressed Sequence Tags, Genes, Genes, Homeobox, Genetic Linkage, Humans, In Situ Hybridization, Fluorescence, Mice, Chemokines, C, Chickens genetics, Chromosomes, E-Selectin genetics, Lymphokines genetics, Sialoglycoproteins genetics
- Abstract
Three differentially expressed selectin genes (SELE, SELP, and SELL), important in the initial stages of leukocyte extravasation, have been reported in mammals. All three genes map close to the chemokine SCYC1 (small inducible cytokine subfamily C, member 1) in a large conserved chromosomal segment that extends from RXRG (retinoic acid receptor, gamma) to TNNT2 (troponin T2) on Chromosome (Chr) 1 in both human and mouse. In the mouse, we demonstrate that Sele is flanked by Prrx1 (paired-related homeobox gene 1) and Scyc1 and define the order of, and distances between, loci as centromere-Prrx1-(0.7+/-0.7 cM)-Sele-(1.2+/-0.9 cM)-Scyc1-telomere. In the chicken, we isolated BAC clones containing PRRX1, SELE, and SCYC1 and positioned them by fluorescent in situ hybridization. SELE and PRRX1 mapped to the short arm of chicken Chr 8 and SCYC1 mapped to the region equivalent to 1q11-1q13 on the long arm of chicken Chr 1. The location of SELE on chicken Chr 8 was independently established by linkage analysis of COM0185, an (AT)16 microsatellite locus identified in a BAC clone that contained SELE. COM0185 was linked to several loci that mapped to one end of chicken Chr 8, with the order of loci, and genetic distances (in cM) between them defined as MSU0435, MSU0325-(7.8+/-3.7)-COM0185-(5.8+/-3.2)-ROS0338-(9.6+/-4.0)-ABR0322-(3.8+/-2.6)-GLUL. We have therefore positioned an evolutionary breakpoint in mammals and chickens between SELE and SCYC1. Furthermore, comparative mapping analysis of the RXRG-TNNT2 chromosomal segment that is conserved on human and mouse Chr 1 indicates that it is divided into four segments in the chicken, each of which maps to a different chromosome.
- Published
- 2001
- Full Text
- View/download PDF
41. The OMP-lacZ transgene mimics the unusual expression pattern of OR-Z6, a new odorant receptor gene on mouse chromosome 6: implication for locus-dependent gene expression.
- Author
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Pyrski M, Xu Z, Walters E, Gilbert DJ, Jenkins NA, Copeland NG, and Margolis FL
- Subjects
- Animals, Axons metabolism, Chromosome Mapping, Chromosomes, Human, Pair 7 genetics, Genomic Library, Humans, In Situ Hybridization, Mice, Mice, Inbred Strains, Mice, Transgenic, Molecular Sequence Data, Multigene Family, Nerve Tissue Proteins genetics, Olfactory Bulb cytology, Olfactory Marker Protein, Olfactory Mucosa cytology, Olfactory Mucosa innervation, Olfactory Mucosa metabolism, Olfactory Receptor Neurons cytology, Olfactory Receptor Neurons metabolism, Organ Specificity, Promoter Regions, Genetic, Sequence Homology, Nucleic Acid, beta-Galactosidase biosynthesis, beta-Galactosidase genetics, Chromosomes genetics, Gene Expression Regulation physiology, Genes, Reporter, Receptors, Odorant biosynthesis, Receptors, Odorant genetics, Transgenes
- Abstract
Reporter gene expression in the olfactory epithelium of H-lacZ6 transgenic mice mimics the cell-selective expression pattern known for some odorant receptor genes. The transgene construct in these mice consists of the lacZ coding region, driven by the proximal olfactory marker protein (OMP) gene promoter, and shows expression in a zonally confined subpopulation of olfactory neurons. To address mechanisms underlying the odorant receptor-like expression pattern of the lacZ construct, we analyzed the transgene-flanking region and identified OR-Z6, the first cloned odorant receptor gene that maps to mouse chromosome 6. OR-Z6 bears the highest sequence similarity (85%) to a human odorant receptor gene at the syntenic location on human chromosome 7. We analyzed the expression pattern of OR-Z6 in olfactory tissues of H-lacZ6 mice and show that it bears strong similarities to that mapped for beta-galactosidase. Expression of both genes in olfactory neurons is primarily restricted to the same medial subregion of the olfactory epithelium. Axons from both neuronal subpopulations project to the same ventromedial aspect of the anterior olfactory bulbs. Furthermore, colocalization analyses in H-lacZ6 mice demonstrate that OR-Z6-reactive glomeruli receive axonal input from lacZ-positive neurons as well. These results suggest that the expression of both genes is coordinated and that transgene expression in H-lacZ6 mice is regulated by locus-dependent mechanisms.
- Published
- 2001
42. Genomic organization of the Shc-related phosphotyrosine adapters and characterization of the full-length Sck/ShcB: specific association of p68-Sck/ShcB with pp135.
- Author
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Kojima T, Yoshikawa Y, Takada S, Sato M, Nakamura T, Takahashi N, Copeland NG, Gilbert DJ, Jenkins NA, and Mori N
- Subjects
- Amino Acid Sequence, Animals, Base Sequence, Cell Line, Cloning, Molecular, Crosses, Genetic, Exons, Female, Humans, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Multigene Family, Muridae, Nerve Tissue Proteins chemistry, Nerve Tissue Proteins metabolism, Phosphotyrosine metabolism, Proteins chemistry, Proteins metabolism, Rats, Recombinant Proteins metabolism, Reverse Transcriptase Polymerase Chain Reaction, Sequence Alignment, Sequence Deletion, Sequence Homology, Amino Acid, Shc Signaling Adaptor Proteins, Src Homology 2 Domain-Containing, Transforming Protein 1, Src Homology 2 Domain-Containing, Transforming Protein 2, Src Homology 2 Domain-Containing, Transforming Protein 3, Transfection, src Homology Domains, Adaptor Proteins, Signal Transducing, Chromosome Mapping, Nerve Tissue Proteins genetics, Neuropeptides, Proteins genetics
- Abstract
The Shc gene family is an emerging family, containing at least three members designated Shc/ShcA, Sck/Sli/ShcB, N-Shc/Rai/ShcC in mammals. In this study, we determined the genomic organization of the mouse Shc family. Coding regions of ShcA, B, and C each comprised 12 exons, spanned approximately 6, 20, and 65 kb, and located on chromosome 3, 10, and 13, respectively. Based on this genome analysis, we determined the full-length structure of mouse Sck/ShcB as a 68-kD protein. We found that the 68-kD full-length Sck/ShcB was more efficiently phosphorylated upon EGF treatment than the previously-analyzed CH2-deleted form. We also found that Sck specifically interacted with a 135-kD phosphoprotein (pp135) through its SH2 domain following membrane depolarization. The Sck-pp135 interaction was reduced by Src kinase inhibitors. These results suggest that Sck, but not N-Shc nor Shc, transmit signals in conjunction with pp135 following Src activation and/or calcium entry in the cell., (Copyright 2001 Academic Press.)
- Published
- 2001
- Full Text
- View/download PDF
43. Sequence, chromosomal location and expression analysis of the murine homologue of human RAD51L2/RAD51C.
- Author
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Leasure CS, Chandler J, Gilbert DJ, Householder DB, Stephens R, Copeland NG, Jenkins NA, and Sharan SK
- Subjects
- Animals, Base Sequence, Blotting, Northern, Chromosome Mapping, Cloning, Molecular, DNA, Complementary chemistry, DNA, Complementary genetics, Female, Gene Expression, Gene Expression Regulation, Developmental, Humans, Male, Mice, Mice, Inbred C57BL, Mice, Inbred Strains, Molecular Sequence Data, Muridae, RNA, Messenger genetics, RNA, Messenger metabolism, Rad51 Recombinase, Sequence Alignment, Sequence Analysis, DNA, Sequence Homology, Amino Acid, Testis metabolism, Tissue Distribution, DNA-Binding Proteins genetics
- Abstract
The Rad51 protein has been shown to play a vital role in the DNA repair process. In humans, its interaction with proteins like BRCA1 and BRCA2 has provided an insight into the mechanism of how these molecules function as tumor suppressors. Several members of the Rad51-like family have been recently identified, including RAD51L2. This gene has been found to be amplified in breast tumors suggesting its role in tumor progression. Here, we describe the cloning of the murine homologue of the human RAD51L2/RAD51C gene. Sequence analysis has revealed that the murine Rad51l2 protein is 86% identical and 93% similar to its human homologue. In spite of such high sequence conservation, the murine protein lacks the first nine amino acids present in the human protein. We have cloned and confirmed the sequence of the 5' end of the murine Rad51l2 cDNA using 5' RACE technique as well as by sequencing the genomic region flanking the first exon of the murine Rad51l2 gene. Northern analysis shows that Rad51l2 is expressed in several adult tissues as well as in embryos at various developmental stages. The murine Rad51l2 gene maps to chromosome 11 and is located in the syntenic region of human chromosome 17q22-23, where the human RAD51L2 is present.
- Published
- 2001
- Full Text
- View/download PDF
44. Promoter function of the angiogenic inducer Cyr61gene in transgenic mice: tissue specificity, inducibility during wound healing, and role of the serum response element.
- Author
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Latinkic BV, Mo FE, Greenspan JA, Copeland NG, Gilbert DJ, Jenkins NA, Ross SR, and Lau LF
- Subjects
- Animals, Chromosome Mapping, Crosses, Genetic, Cysteine-Rich Protein 61, Fibroblasts physiology, Gene Expression, Growth Substances physiology, Immediate-Early Proteins physiology, Immunohistochemistry, In Situ Hybridization, Mice, Mice, Inbred C57BL, Mice, Transgenic, Neovascularization, Physiologic, beta-Galactosidase genetics, Growth Substances genetics, Immediate-Early Proteins genetics, Intercellular Signaling Peptides and Proteins, Organ Specificity, Promoter Regions, Genetic, Response Elements, Wound Healing
- Abstract
The cysteine-rich angiogenic protein 61 (Cyr61) is an extracellular matrix-associated, heparin-binding protein that mediates cell adhesion, stimulates cell migration, and enhances growth factor-induced cell proliferation. Cyr61 also promotes chondrogenic differentiation and induces neovascularization. In this study, we show that a 2-kb fragment of the Cyr61 promoter, which confers growth factor-inducible expression in cultured fibroblasts, is able to drive accurate expression of the reporter gene lacZ in transgenic mice. Thus, transgene expression was observed in the developing placenta and embryonic cardiovascular, skeletal, and central and peripheral nervous systems. The sites of transgene expression are consistent with those observed of the endogenous Cyr61 gene as determined by in situ hybridization and immunohistochemistry. The transgene expression in the cardiovascular system does not require the serum response element, a promoter sequence essential for transcriptional activation of Cyr61 by serum growth factors in cultured fibroblasts. Because the serum response element contains the CArG box, a sequence element implicated in cardiovascular-specific gene expression, the nonessential nature of this sequence for cardiovascular expression of Cyr61 is unexpected. Furthermore, the Cyr61 promoter-driven lacZ expression is inducible in granulation tissue during wound healing, as is synthesis of the endogenous Cyr61 protein, suggesting a role for Cyr61 in wound healing. Consistent with this finding, purified Cyr61 protein promotes the healing of a wounded fibroblast monolayer in culture. In addition, we mapped the mouse Cyr61 gene to the distal region of chromosome 3. Together, these results define the functional Cyr61 promoter in vivo, and suggest a role of Cyr61 in wound healing through its demonstrated angiogenic activities upon endothelial cells and its chemotactic and growth promoting activities upon fibroblasts.
- Published
- 2001
- Full Text
- View/download PDF
45. The gene for a variant form of the polyadenylation protein CstF-64 is on chromosome 19 and is expressed in pachytene spermatocytes in mice.
- Author
-
Dass B, McMahon KW, Jenkins NA, Gilbert DJ, Copeland NG, and MacDonald CC
- Subjects
- Amino Acid Sequence, Animals, Antibodies, Monoclonal immunology, DNA, Complementary isolation & purification, Humans, Male, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Peptide Mapping, RNA metabolism, RNA-Binding Proteins immunology, mRNA Cleavage and Polyadenylation Factors, Chromosome Mapping, RNA-Binding Proteins genetics, Spermatocytes metabolism
- Abstract
Many mRNAs in male germ cells lack the canonical AAUAAA but are normally polyadenylated (Wallace, A. M., Dass, B., Ravnik, S. E., Tonk, V., Jenkins, N. A., Gilbert, D. J., Copeland, N. G., and MacDonald, C. C. (1999) Proc. Natl. Acad Sci. U. S. A. 96, 6763-6768). Previously, we demonstrated the presence of two distinct forms of the M(r) 64,000 protein of the cleavage stimulation factor (CstF-64) in mouse male germ cells and in brain, a somatic M(r) 64,000 form and a variant M(r) 70,000 form. The variant form was specific to meiotic and postmeiotic germ cells. We localized the gene for the somatic CstF-64 to the X chromosome, which would be inactivated during male meiosis. This suggested that the variant CstF-64 was an autosomal homolog activated during that time. We have named the variant form "tau CstF-64," and we describe here the cloning and characterization of the mouse tauCstF-64 cDNA, which maps to chromosome 19. The mouse tauCstF-64 protein fits the criteria of the variant CstF-64, including antibody reactivity, size, germ cell expression, and a common proteolytic digest pattern with tauCstF-64 from testis. Features of mtauCstF-64 that might allow it to promote the germ cell pattern of polyadenylation include a Pro --> Ser substitution in the RNA-binding domain and significant changes in the region that interacts with CstF-77.
- Published
- 2001
- Full Text
- View/download PDF
46. Comparison of pretransplant transfusion with or without cyclosporin inhibition on post transplant outcome.
- Author
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Pietra B, Ivy DD, Sondheimer HM, Shaffer EM, Mashburn C, Ripe DW, Gilbert DJ, Kyle TE, Mitchell MB, Campbell DN, and Boucek MM
- Published
- 2001
- Full Text
- View/download PDF
47. Transplant coronary artery disease in pediatrics: favorable outcome with medical therapy.
- Author
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Boucek MM, Sondheimer HM, Ivy DD, Shaffer EM, Mashburn C, Ripe DW, Gilbert DJ, Kyle TE, Campbell DN, and Pietra B
- Published
- 2001
- Full Text
- View/download PDF
48. A novel transgenic marker for migrating limb muscle precursors and for vascular smooth muscle cells.
- Author
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Tidhar A, Reichenstein M, Cohen D, Faerman A, Copeland NG, Gilbert DJ, Jenkins NA, and Shani M
- Subjects
- Animals, Arteries injuries, Blotting, Southern, Cell Division, Cell Movement, Chromosomes, Down-Regulation, Ectoderm metabolism, Embryo, Mammalian metabolism, Expressed Sequence Tags, Extremities embryology, Genes, Reporter, Immunohistochemistry, Lac Operon, Male, Mice, Mice, Inbred C57BL, Mice, Transgenic, Models, Genetic, Muscle, Skeletal embryology, Muscle, Smooth embryology, Polymerase Chain Reaction, Reverse Transcriptase Polymerase Chain Reaction, Time Factors, Tissue Distribution, Wound Healing, beta-Galactosidase metabolism, Muscle, Smooth, Vascular embryology, Muscles embryology, Transgenes
- Abstract
A unique pattern of LacZ expression was found in a transgenic mouse line, likely due to regulatory elements at the site of integration. Two new genes flanking the transgene were identified. At early stages of development, the transgene is transiently expressed in ventro-lateral demomyotomal cells migrating from the somites into the limb buds. At late developmental stages and in the adult, lacZ staining marks vascular smooth muscle cells throughout the vascular bed, with the exception of the major elastic arteries, and in pericytes. No expression was detected in skeletal and smooth muscles. Different patterns of expression in vascular smooth muscles was observed at distinct levels of the vascular tree, in arteries as well as in veins. Vessel injury, resulting in stimulation of smooth muscle cells proliferation and migration, is associated with transgene down-regulation. After the formation of neointima thickening, it is reactivated. This transgenic insertion may therefore be used as a useful marker to identify novel physiological cues or genetic elements involved in the regulation of the vascular smooth muscle phenotype(s). It may also provide an experimental tool for studying vasculature and the involvement of pericytes in regulating microvascular homeostasis., (Copyright 2001 Wiley-Liss, Inc.)
- Published
- 2001
- Full Text
- View/download PDF
49. Structure and chromosome location of the mouse P2X(1) purinoceptor gene (P2rx1).
- Author
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Liang SX, Jenkins NA, Gilbert DJ, Copeland NG, and Phillips WD
- Subjects
- Animals, Base Sequence, Cloning, Molecular, Conserved Sequence genetics, Crosses, Genetic, Exons genetics, Female, Introns genetics, Lod Score, Male, Mice, Mice, Inbred C57BL, RNA Splice Sites genetics, RNA, Messenger analysis, RNA, Messenger genetics, Receptors, Purinergic P2X, Chromosome Mapping, Receptors, Purinergic P2 genetics
- Abstract
P2X(1) receptors are ATP-gated cation channels that mediate the fast, purinergic component of sympathetic nerve-smooth muscle neurotransmission in the mouse vas deferens and may serve comparable functions in the urinary bladder and the arteries. The gene for mouse P2X(1) (P2rx1) was cloned and its genomic structure defined by sequencing. The gene spans about 10 kb and consists of 12 exons. All splice sites conformed to the GT-AG motif and the exon-intron boundaries were largely conserved with other members of the P2X gene family so far cloned. A single transcription-starting site was identified by 5' RACE analysis, 233 bp upstream of the translation start site. The P2X(1) gene maps to the central region of mouse chromosome 11., (Copyright 2001 S. Karger AG, Basel.)
- Published
- 2001
- Full Text
- View/download PDF
50. Cloning, expression analysis, and chromosomal localization of murine and human homologues of a Xenopus mix gene.
- Author
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Robb L, Hartley L, Begley CG, Brodnicki TC, Copeland NG, Gilbert DJ, Jenkins NA, and Elefanty AG
- Subjects
- Amino Acid Sequence, Animals, Blotting, Northern, Chromosome Mapping, Chromosomes, Human, Pair 1 genetics, Cloning, Molecular, Embryo, Mammalian anatomy & histology, Gene Expression, Genes, Homeobox genetics, Homeodomain Proteins analysis, Homeodomain Proteins chemistry, Humans, In Situ Hybridization, Mice, Mice, Inbred C57BL, Molecular Sequence Data, RNA, Messenger analysis, Reverse Transcriptase Polymerase Chain Reaction, Sequence Alignment, Trans-Activators analysis, Trans-Activators chemistry, Embryo, Mammalian physiology, Homeodomain Proteins genetics, Trans-Activators genetics, Xenopus Proteins
- Abstract
We report the cloning and chromosomal localization of murine and human Mix genes, members of a subclass of paired-like homeobox genes of which the Xenopus laevis Mix.1 gene is the founding member. The murine Mix gene was mapped to the distal region of chromosome 1 and the human region to the syntenic region 1q41-42. Northern analysis and RT-PCR of murine adult and embryonic tissues demonstrated that Mix expression was restricted to the early embryo. Whole-mount in situ hybridization revealed patchy but symmetrical Mix expression in visceral endoderm of embryonic day (E)5.5 embryos. In slightly older embryos, the expression was skewed to one side of the embryo and by E6.5, at the onset of gastrulation, expression was seen in the epiblast, visceral endoderm, nascent mesoderm, and the primitive streak. This expression pattern was maintained in mid- and late-streak embryos. In early bud-stage embryos, expression was strongest in the proximal two thirds of the streak, extending to the base of the allantois. By the headfold-stage, expression was confined to the remnant of the primitive streak in the caudal region of the embryo and, after E8.0, in the caudal notochord and tail bud mesoderm. Mix transcripts were no longer detectable after embryonic day 9.5., (Copyright 2000 Wiley-Liss, Inc.)
- Published
- 2000
- Full Text
- View/download PDF
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