163 results on '"Follitropin"'
Search Results
2. Heterologous Production and Glycosylation of Japanese Eel Follitropin Using Silkworm
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Kwan Sik Min, Jae Man Lee, Sun Jung Jo, Dae-Jung Kim, Takahiro Kusakabe, Ji Hyun Choi, and Sun Mee Hong
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0106 biological sciences ,endocrine system ,Glycosylation ,medicine.drug_class ,Biomedical Engineering ,Heterologous ,Bioengineering ,Follitropin ,Biology ,01 natural sciences ,Applied Microbiology and Biotechnology ,03 medical and health sciences ,chemistry.chemical_compound ,010608 biotechnology ,medicine ,Japanese eel ,Equine chorionic gonadotropin ,030304 developmental biology ,0303 health sciences ,Hormone activity ,fungi ,biology.organism_classification ,chemistry ,Biochemistry ,Vitellogenesis ,Gonadotropin ,Biotechnology - Abstract
Follitropin, an important gonadotropin hormone, participates in vitellogenesis and spermatogenesis. Equine chorionic gonadotropin (eCG) can induce gonadotropin hormone activity in non-equid species and exhibits a long biological half-life. Here, we report the production, using silkworm larval and pupal systems, of biologically active recombinant hybrid-type follitropins based on the coding sequence of the eCG C-terminal peptide (CTP) between the mature β- and α-chains of eel. The three constructs, rJeFSH, rJeFSH·eCG, and rJeFSH·2xeCG were produced and verified to be N- or O-glycosylated and secreted mature peptides. Although rJeFSH·eCG contains more elaborate O-linked carbohydrate chains than rJeFSH, it elicited no significant in vitro oocyte maturation, which may be a result of insufficient terminal sialylation of its N-and O-linked carbohydrate chains. Then, a hybrid of rJeFSH·2xeCG extended with two eCG CTP. Furthermore, the receptor binding assay revealed potency of rJeFSH and rJeFSH·2xeCG to be a few folds greater than that of rJeFSH·eCG. The findings of this study will be useful for the development of more efficient GTHs in teleosts, including eels, when various modifications with two or more extended eCG CTP produced by silkworm are included.
- Published
- 2019
3. New Human Follitropin Preparations: How Glycan Structural Differences May Affect Biochemical and Biological Function and Clinical Effect
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Alfredo Ulloa-Aguirre and James A. Dias
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0301 basic medicine ,glycoprotein ,Male ,Glycan ,endocrine system ,Glycosylation ,Acetylgalactosamine ,Endocrinology, Diabetes and Metabolism ,pharmocodynamics ,Follitropin ,Context (language use) ,Review ,Asialoglycoprotein Receptor ,CHO Cells ,lcsh:Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,0302 clinical medicine ,Endocrinology ,Cricetulus ,Polysaccharides ,therapeutics ,Animals ,Humans ,Protein Isoforms ,Glycoproteins ,chemistry.chemical_classification ,lcsh:RC648-665 ,030219 obstetrics & reproductive medicine ,biology ,N-Acetylneuraminic Acid ,Recombinant Proteins ,Sialic acid ,Rats ,030104 developmental biology ,chemistry ,Biochemistry ,Liver ,Galactosamine ,biology.protein ,Asialoglycoprotein receptor ,Female ,Follicle Stimulating Hormone, Human ,Follicle Stimulating Hormone ,Glycoprotein ,follitropin ,pharmacokinetics - Abstract
It is well accepted that pituitary follitropin is secreted into the circulation as a mixture of variants, which differ not in primary structure but rather at the level of glycosylation. These glycosidic forms vary in the number of glycosylation sites filled, complexity of glycosidic chains, and sialylation and sulfation. It is generally agreed that high sialylation, 2,3 sialic acid capping of terminal N-acetyl galactosamine or galactose leads to longer circulating half-life, by blocking binding of asialoglycoprotein receptor (ASGPR) in the liver. In contrast, 2,6 sialic acid found in humans does not prevent recognition of galactose and N-acetyl galactosamine by ASGPR. Few studies on clinical outcomes comparing differences in sialylation of follitropin found in commercially available preparations are available. Thus, there is a clear need for a consortium of open data to address this unmet need. Recently, FSH glycosylation, primarily on the β-subunit, which varies as women age, has emerged as a key modifier of follitropin action, with profound biological effects in vivo in animal models. To date, limited information of recombinant follitropin hormone preparations is available. Thus, most of the studies with FSH that is well characterized biochemically have been done in vitro, with engineered non gonadal host cells bearing recombinant receptors or in animal models. Since limited studies in human granulosa cells are available, a question is whether structural differences in glycosylation in commercially available follitropin affects biological function and clinical effect in humans. The presence of fucose, for example, has not been studied greatly even though, in the case of antibody therapy it has been shown to have a large effect on antibody targeting. This review on glycosidic variability of follitropin from the biochemical/structural point of view reflects on this question and presents an assessment in the context of available published data. If clinical differences are to be expected or not, the readers will have a better understanding of the evidence for and limitations of such expectations.
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- 2021
4. Gonadotropin Hormones and Their Receptors
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James A. Dias, Prema Narayan, and Alfredo Ulloa-Aguirre
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endocrine system ,medicine.medical_specialty ,urogenital system ,medicine.drug_class ,Follitropin ,Ovary ,Biology ,Human chorionic gonadotropin ,Syncytiotrophoblast ,medicine.anatomical_structure ,Endocrinology ,Internal medicine ,medicine ,Gonadotropin ,Receptor ,Luteinizing hormone ,reproductive and urinary physiology ,hormones, hormone substitutes, and hormone antagonists ,Hormone - Abstract
The gonads (ovary, testis) produce gametes (oocytes and spermatozoa). The pituitary glycoprotein hormones luteinizing hormone (LH, lutropin) and follicle-stimulating hormone (FSH, follitropin) are gonadotropins because their targets are the gonads. Another hormone produced by the syncytiotrophoblast of the human placenta is human chorionic gonadotropin (hCG, choriogonadotropin), and its activity is like LH. This review describes the protein structure, gene structure, and regulation of biosynthesis of both the gonadotropins and their membrane bound receptors. The functions of both hormones and receptors are covered extensively, as well as naturally occurring mutations that have been identified in disease states and therapies derived from their study.
- Published
- 2019
5. DOSE ADJUSTMENT OF FOLLICLE-STIMULATING HORMONE (FSH) DURING OVARIAN STIMULATION AS PART OF MEDICALLY-ASSISTED REPRODUCTION IN CLINICAL STUDIES: A SYSTEMATIC REVIEW COVERING 10 YEARS (2007–2017)
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Georg Griesinger, Xiaoyan Yin, Wilma Bilger, Salvatore Longobardi, M.C. Mahony, Deborah Denis, Thomas D'Hooghe, Antonio La Marca, and Human M. Fatemi
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QH471-489 ,POOR RESPONDERS ,LIVE BIRTH-RATES ,medicine.medical_treatment ,Medically-assisted reproduction ,HYPERSTIMULATION SYNDROME ,Stimulation ,Review ,WOMEN STARTING IVF/ICSI ,Follicle-stimulating hormone ,Endocrinology ,Controlled ovarian stimulation (COS) ,media_common ,Clinical Trials as Topic ,Reproductive Biology ,biology ,Reproduction ,Incidence (epidemiology) ,Obstetrics and Gynecology ,Anti-Müllerian hormone ,Embryo transfer ,Recombinant Proteins ,ALPHA PEN INJECTOR ,Female ,Gonadotropin ,Life Sciences & Biomedicine ,medicine.medical_specialty ,Reproductive Techniques, Assisted ,Follicle-stimulating hormone (FSH) ,medicine.drug_class ,ANTI-MULLERIAN HORMONE ,media_common.quotation_subject ,EMBRYO-TRANSFER ,Ovarian Hyperstimulation Syndrome ,Endocrinology & Metabolism ,Ovulation Induction ,Internal medicine ,medicine ,Humans ,FOLLITROPIN ALPHA ,Assisted reproductive technology (ART) treatment ,Follitropin ,FSH dose adjustment ,In vitro fertilization (IVF) ,Recombinant-human FSH ,Systematic review ,In vitro fertilisation ,Science & Technology ,Assisted reproductive technology ,Dose-Response Relationship, Drug ,Drug Tapering ,business.industry ,Gynecology and obstetrics ,Confidence interval ,Clinical trial ,Reproductive Medicine ,biology.protein ,RG1-991 ,GNRH ANTAGONIST ,Follicle Stimulating Hormone ,IN-VITRO FERTILIZATION ,business ,Developmental Biology - Abstract
Background Individualization of the follicle-stimulating hormone (FSH) starting dose is considered standard clinical practice during controlled ovarian stimulation (COS) in patients undergoing assisted reproductive technology (ART) treatment. Furthermore, the gonadotropin dose is regularly adjusted during COS to avoid hyper- or hypo-ovarian response, but limited data are currently available to characterize such adjustments. This review describes the frequency and direction (increase/decrease) of recombinant-human FSH (r-hFSH) dose adjustment reported in clinical trials. Methods We evaluated the proportion of patients undergoing ART treatment who received ≥ 1 r-hFSH dose adjustments. The inclusion criteria included studies (published Sept 2007 to Sept 2017) in women receiving ART treatment that allowed dose adjustment within the study protocol and that reported ≥ 1 dose adjustments of r-hFSH; studies not allowing/reporting dose adjustment were excluded. Data on study design, dose adjustment and patient characteristics were extracted. Point-incidence estimates were calculated per study and overall based on pooled number of cycles with dose adjustment across studies. The Clopper–Pearson method was used to calculate 95% confidence intervals (CI) for incidence where adjustment occurred in Results Initially, 1409 publications were identified, of which 318 were excluded during initial screening and 1073 were excluded after full text review for not meeting the inclusion criteria. Eighteen studies (6630 cycles) reported dose adjustment: 5/18 studies (1359 cycles) reported data for an unspecified dose adjustment (direction not defined), in 10/18 studies (3952 cycles) dose increases were reported, and in 11/18 studies (5123 cycles) dose decreases were reported. The studies were performed in women with poor, normal and high response, with one study reporting in oocyte donors and one in obese women. The median day that dose adjustment was permitted was Day 6 after the start of treatment. The point estimates for incidence (95% CI) for unspecified dose adjustment, dose increases, and dose decreases were 45.3% (42.7, 48.0), 19.2% (18.0, 20.5), and 9.5% (8.7, 10.3), respectively. Conclusions This systematic review highlights that, in studies in which dose adjustment was allowed and reported, the estimated incidence of r-hFSH dose adjustments during ovarian stimulation was up to 45%.
- Published
- 2020
6. Follicle Stimulating Hormone Receptor Signaling is Regulated by Lipid Raft Residency
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Brian D. Cohen and Megan M. Wells
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medicine.medical_specialty ,Follitropin ,Biology ,Biochemistry ,Follicle-stimulating hormone ,Endocrinology ,Internal medicine ,Genetics ,medicine ,Follicle-stimulating hormone receptor ,Hormone signaling ,Molecular Biology ,Lipid raft ,Biotechnology - Published
- 2018
7. Occurrence of ovarian follicular dominance during stimulation for IVM impacts usable blastocyst yield
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Ricardo Pella, Patricia Orihuela, Francisco Escudero, Sergio Romero, Ygor Pérez, and Mario García
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0301 basic medicine ,blastocysts ,Pregnancy Rate ,retrospective study ,0302 clinical medicine ,Human fertilization ,Oogenesis ,Ovarian Follicle ,Pregnancy ,Follicular phase ,clinical article ,030219 obstetrics & reproductive medicine ,adult ,Blastocyst Transfer ,follicular dominance ,Embryo ,Polycystic ovary ,embryo culture ,female ,medicine.anatomical_structure ,IVM ,embryonic structures ,luteinizing hormone ,PCO ,Female ,Original Article ,follitropin ,Infertility, Female ,Polycystic Ovary Syndrome ,Adult ,endocrine system ,Fertilization in Vitro ,Biology ,Article ,Andrology ,oocyte retrieval ,03 medical and health sciences ,Ovulation Induction ,estradiol ,ovary polycystic disease ,medicine ,Humans ,controlled study ,human ,Blastocyst ,gonadotropin ,embryo transfer ,Retrospective Studies ,pregnancy outcome ,blastocyst ,urogenital system ,embryo development ,in vitro oocyte maturation ,medicine.disease ,Oocyte ,Embryo Transfer ,In Vitro Oocyte Maturation Techniques ,030104 developmental biology ,fertilization ,Oocytes - Abstract
Objective To evaluate the influence of ovarian follicular dominance on the outcome of oocyte in-vitro maturation. Methods This retrospective cohort study included 21 patients with polycystic ovaries or polycystic ovary syndrome (Rotterdam criteria, 2004) subjected to 24 invitro maturation (IVM) cycles between October 2015 and January 2017. Patients undergoing IVM received minimal gonadotropin stimulation starting on day 2 or 3 of the cycle; ovum pick-up typically occurred on days 6 to 8. No hCG-trigger shot was given. Following 30h of IVM, mature oocytes were inseminated by ICSI and the resulting embryos cultured up to the blastocyst stage. Results Ovarian follicular dominance was observed in nine of the 24 IVM cycles. Oocyte IVM yielded an overall maturation rate of 69.3±23.8%, and no difference was observed when the groups with or without a dominant follicle were assessed independently. The rates of fertilization and usable blastocysts per fertilized oocyte, mature oocyte (Metaphase II) or cumulus-oocyte-complex were nearly three times higher (28.7±22.5%) in the group without ovarian follicular dominance. No differences were found in the clinical pregnancy rates attained by the individuals with or without a dominant follicle after 21 vitrified-warmed blastocyst transfer cycles. Conclusion Occurrence of ovarian follicular dominance during hormonal stimulation for in-vitro maturation negatively impacted embryological outcomes. Strategies devised to limit the appearance of ovarian follicular dominance must be further explored.
- Published
- 2018
8. The Integrated Hypothalamic Tachykinin-Kisspeptin System as a Central Coordinator for Reproduction
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Stephanie B. Seminara, Oline K. Rønnekleiv, Silvia Leon, Martha A. Bosch, Rona S. Carroll, Cadence True, Manuel Tena-Sempere, Leonor Pinilla, Ursula B. Kaiser, Víctor M. Navarro, and Serap Simavli
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Male ,neurokinin 1 receptor ,substance P ,Neurokinin A ,Substance P ,animal cell ,Gonadotropin-releasing hormone ,luteinizing hormone release ,thalamus ventral nucleus ,Receptors, G-Protein-Coupled ,Gonadotropin-Releasing Hormone ,chemistry.chemical_compound ,Endocrinology ,Kisspeptin ,arcuate nucleus ,animal ,hypothalamus ,Mice, Knockout ,Kisspeptins ,Estradiol ,messenger RNA ,C57BL mouse ,adult ,Reproduction ,neurokinin 3 receptor ,respiratory system ,neurokinin 2 receptor ,female ,priority journal ,Hypothalamus ,luteinizing hormone ,Neurokinin B ,secretion (process) ,follitropin ,hormones, hormone substitutes, and hormone antagonists ,gonadorelin release ,hormonal regulation ,Agonist ,medicine.medical_specialty ,tachykinin receptor ,gonadotropin release ,medicine.drug_class ,animal experiment ,G protein coupled receptor ,Kiss1 protein, mouse ,Biology ,Article ,animal tissue ,kisspeptin ,Arcuate nucleus ,Internal medicine ,medicine ,Animals ,controlled study ,gonadotropin ,protein expression ,mouse ,Receptors, Tachykinin ,nonhuman ,Kiss1r protein, mouse ,animal model ,Neuroendocrinology ,Mice, Inbred C57BL ,chemistry ,gonadorelin ,agonists ,Follicle Stimulating Hormone ,knockout mouse ,metabolism ,Receptors, Kisspeptin-1 - Abstract
Tachykinins are comprised of the family of related peptides, substance P (SP), neurokinin A (NKA), and neurokinin B (NKB). NKB has emerged as regulator of kisspeptin release in the arcuate nucleus (ARC), whereas the roles of SP and NKA in reproduction remain unknown. This work explores the roles of SP and NKA in the central regulation of GnRH release. First, central infusion of specific agonists for the receptors of SP (neurokinin receptor 1, NK1R), NKA (NK2R) and NKB (NK3R) each induced gonadotropin release in adult male and ovariectomized, estradiol-replaced female mice, which was absent in Kiss1r-/- mice, indicating a kisspeptin-dependent action. The NK2R agonist, however, decreasedLHrelease in ovariectomized-sham replaced females, as documented forNK3R agonists but in contrast to the NK1R agonist, which further increased LH release. Second, Tac1 (encoding SP and NKA) expression in the ARC and ventromedial nucleus was inhibited by circulating estradiol but did not colocalize with Kiss1 mRNA. Third, about half of isolated ARC Kiss1 neurons expressed Tacr1 (NK1R) and 100% Tacr3 (NK3R); for anteroventral-periventricular Kiss1 neurons andGnRHneurons, approximately one-fourth expressed Tacr1 and one-tenth Tacr3; Tacr2 (NK2R) expression was absent in all cases. Overall, these results identify a potent regulation of gonadotropin release by the SP/NK1R and NKA/NK2R systems in the presence of kisspeptin-Kiss1r signaling, indicating that they may, along with NKB/NK3R, control GnRH release, at least in part through actions on Kiss1 neurons. Copyright © 2015 by the Endocrine Society.
- Published
- 2014
9. New mutations in non-syndromic primary ovarian insufficiency patients identified via whole-exome sequencing
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Manuel A. Patarroyo, Jacques Young, July Constanza Buitrago, Liliana C Patiño, Brigitte Delemer, Nadine Binart, Isabelle Beau, Carlos F. Suárez, Paul Laissue, Ronald Gonzalez, and Carolina Carlosama
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Luteinizing hormone ,Signal peptide ,single nucleotide ,Granulosa cell ,Genome-wide association study ,Gene mutation ,Primary Ovarian Insufficiency ,Gene ,0302 clinical medicine ,Models ,Protein stability ,Cell differentiation ,Ovary insufficiency ,Referral and Consultation ,Exome sequencing ,Cell proliferation ,Expert system ,030219 obstetrics & reproductive medicine ,Estradiol ,Oocitos ,Molecular etiology ,Polygenic disease ,Bmpr1b gene ,Patient referral ,Bone morphogenetic protein receptors ,Multicenter study ,Clinical trial ,Retrospective study ,Intercellular Signaling Peptides and Proteins ,Cohort analysis ,Human ,Bioinformatics ,Single-nucleotide polymorphism ,Expert Systems ,Major clinical study ,Primary ovarian insufficiency ,Molecular dynamics ,Article ,03 medical and health sciences ,Molecular dynamics simulation ,Exome Sequencing ,Genetic predisposition ,Frameshift mutation ,Genetics ,Humans ,Polymorphism ,Biology ,Premature ovarian failure ,Gene mapping ,Bone Morphogenetic Protein Receptors, Type I ,Retrospective Studies ,Granulosa cell differentiation ,Sustitución de Aminoácidos ,Whole exome sequencing ,Computational Biology ,Gene frequency ,Retrospective studies ,030104 developmental biology ,Granullosa Cells ,Reproductive Medicine ,Células de la Granulosa ,Mutation ,Estudios de Cohortes ,0301 basic medicine ,Models, Molecular ,Candidate gene ,Gene locus ,Computational biology ,Cohort Studies ,Expert systems ,Ovarian Follicle ,Ovary follicle development ,Missense mutation ,Female infertility ,Grem1 protein ,Sanger sequencing ,Nonsense mutation ,Protein Stability ,Folículo Ovárico ,Rehabilitation ,Obstetrics and Gynecology ,Referral and consultation ,Bone morphogenetic protein receptor 1 ,Meiosis ,Chemistry ,Whole-exome sequencing ,symbols ,Female ,Intercellular signaling peptides and proteins ,France ,Bmpr1b protein ,Ovulation ,Adult ,Adolescent ,Receptores de Proteínas Morfogenéticas Óseas de Tipo 1 ,Genetic predisposition to disease ,Molecular Dynamics Simulation ,Polymorphism, Single Nucleotide ,Molecular model ,symbols.namesake ,Young Adult ,Genetic Predisposition to Disease ,molecular ,Grem1 gene ,type i ,Follitropin ,Single nucleotide polymorphism ,Young adult ,Metabolism ,Amino Acid Substitution ,Oocytes ,Genetic variability ,Genome-Wide Association Study - Abstract
STUDY QUESTION Is it possible to identify new mutations potentially associated with non-syndromic primary ovarian insufficiency (POI) via whole-exome sequencing (WES)? SUMMARY ANSWER WES is an efficient tool to study genetic causes of POI as we have identified new mutations, some of which lead to protein destablization potentially contributing to the disease etiology. WHAT IS KNOWN ALREADY POI is a frequently occurring complex pathology leading to infertility. Mutations in only few candidate genes, mainly identified by Sanger sequencing, have been definitively related to the pathogenesis of the disease. STUDY DESIGN, SIZE, DURATION This is a retrospective cohort study performed on 69 women affected by POI. PARTICIPANTS/MATERIALS, SETTING, METHODS WES and an innovative bioinformatics analysis were used on non-synonymous sequence variants in a subset of 420 selected POI candidate genes. Mutations in BMPR1B and GREM1 were modeled by using fragment molecular orbital analysis. MAIN RESULTS AND THE ROLE OF CHANCE Fifty-five coding variants in 49 genes potentially related to POI were identified in 33 out of 69 patients (48%). These genes participate in key biological processes in the ovary, such as meiosis, follicular development, granulosa cell differentiation/proliferation and ovulation. The presence of at least two mutations in distinct genes in 42% of the patients argued in favor of a polygenic nature of POI. LIMITATIONS, REASONS FOR CAUTION It is possible that regulatory regions, not analyzed in the present study, carry further variants related to POI. WIDER IMPLICATIONS OF THE FINDINGS WES and the in silico analyses presented here represent an efficient approach for mapping variants associated with POI etiology. Sequence variants presented here represents potential future genetic biomarkers. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the Universidad del Rosario and Colciencias (Grants CS/CIGGUR-ABN062-2016 and 672-2014). Colciencias supported Liliana Catherine Patiñós work (Fellowship: 617, 2013). The authors declare no conflict of interest. © The Author 2017. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved.
- Published
- 2017
10. A homozygous donor splice-site mutation in the meiotic gene MSH4 causes primary ovarian insufficiency
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Carolina Carlosama, Reiner A. Veitia, Maëva Elzaiat, Heidi Mateus, Paul Laissue, and Liliana C Patiño
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0301 basic medicine ,Luteinizing hormone ,Sanger sequencing ,Unclassified drug ,Karyotype 46 ,Uterus myoma ,Menopause, Premature ,Thyrotropin ,Cell Cycle Proteins ,Gene mutation ,Gene sequence ,Primary Ovarian Insufficiency ,XX ,Gene ,Thyrotropin blood level ,MSH4 protein ,Whole Exome Sequencing ,Cohort Studies ,Exon ,0302 clinical medicine ,Genetics (clinical) ,Exome sequencing ,Priority journal ,Genetics ,030219 obstetrics & reproductive medicine ,Heterozygosity ,Inheritance ,Messenger RNA ,Homozygote ,General Medicine ,Exons ,Complementary DNA ,Premature ovarian failure ,Pedigree ,Early menopause ,Chemistry ,Hela cell line ,Reverse transcription polymerase chain reaction ,Female ,Cohort analysis ,Menopause ,Luteinizing hormone blood level ,Exon skipping ,Human ,Adult ,MSH4 gene ,Heterozygote ,Biology ,Premature ovarian insufficiency ,Secondary amenorrhea ,Article ,03 medical and health sciences ,Exon trapping ,Exome Sequencing ,Case report ,medicine ,Humans ,Genetic variation ,Segregation analysis ,Molecular Biology ,Premature ,Transvaginal echography ,RNA splice site ,Follitropin blood level ,Menarche ,DNA fragment ,Protein ,Follitropin ,medicine.disease ,Gene frequency ,030104 developmental biology ,MSH4 ,Menstrual irregularity ,Metabolism ,Human cell ,Mutation ,RNA Splice Sites ,Controlled study ,Cell cycle protein - Abstract
Premature ovarian insufficiency (POI) is a frequent pathology that affects women under 40 years of age, characterized by an early cessation of menses and high FSH levels. Despite recent progresses in molecular diagnosis, the etiology of POI remains idiopathic in most cases. Whole-exome sequencing of members of a Colombian family affected by POI allowed us to identify a novel homozygous donor splice-site mutation in the meiotic gene MSH4 (MutS Homolog 4). The variant followed a strict mendelian segregation within the family and was absent in a cohort of 135 women over 50 years of age without history of infertility, from the same geographical region as the affected family. Exon trapping experiments showed that the splice-site mutation induced skipping of exon 17. At the protein level, the mutation p.Ile743_Lys785del is predicted to lead to the ablation of the highly conserved Walker B motif of the ATP-binding domain, thus inactivating MSH4. Our study describes the first MSH4 mutation associated with POI and increases the number of meiotic/DNA repair genes formally implicated as being responsible for this condition. © The Author 2017. Published by Oxford University Press. All rights reserved.
- Published
- 2017
11. Congenital Leptin Deficiency and Leptin Gene Missense Mutation Found in Two Colombian Sisters with Severe Obesity
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Monica Alvarez-Jaramillo, Luis G. Celis-Regalado, Carlos Martín Restrepo, Alexandra Arias-Serrano, Angelica M. Garcia-Ordoñez, Shelley A. Cole, Mauricio Arcos-Burgos, Jack W. Kent, Claudio A. Mastronardi, Edna J. Nava-González, Maria E. Yupanqui-Velazco, Carolina Torres-Forero, Julio Licinio, Ernesto Rodríguez-Ayala, Aida P. Giraldo-Peña, Raul A. Bastarrachea, Esteban Medina-Méndez, and Hernan Yupanqui-Lozno
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Luteinizing hormone ,Leptin ,Hirsutism ,Thyrotropin ,Nail hypoplasia ,Dna sequence ,Gene ,Morbid obesity ,Consanguinity ,0302 clinical medicine ,Insulin ,Sleeve gastrectomy ,Child ,High density lipoprotein cholesterol ,Mutation ,Leptin Deficiency ,Estradiol ,digestive, oral, and skin physiology ,Genetic disorder ,LEP gene ,Metformin ,Obesity, Morbid ,Pedigree ,Colombian ,Human ,Pcsk1 gene ,medicine.medical_specialty ,Novel mutation ,Clinical article ,Mc4r gene ,Colombia ,Triacylglycerol ,Congenital leptin deficiency ,Article ,03 medical and health sciences ,consanguinity ,Pomc gene ,Case report ,Genetics ,Humans ,Gene deletion ,Pparg gene ,medicine.disease ,Knee pain ,Extreme obesity ,Obesity ,Prolactin ,Strabismus ,Glucose ,030104 developmental biology ,Endocrinology ,Acne ,Leptin deficiency ,School child ,0301 basic medicine ,Enzyme linked immunosorbent assay ,Walking ,medicine.disease_cause ,Homozygosity ,congenital leptin deficiency ,Exon ,Hemoglobin a1c ,Lep gene ,Missense mutation ,Protein blood level ,Childhood obesity ,extreme obesity ,Amenorrhea ,Genetics (clinical) ,Hypertriglyceridemia ,Point mutation ,Fat mass ,Body weight gain ,Exons ,Body mass ,Telangiectasia ,Female ,hormones, hormone substitutes, and hormone antagonists ,Adult ,Lepr gene ,Adolescent ,lcsh:QH426-470 ,Colombian sisters ,Mutation, Missense ,030209 endocrinology & metabolism ,Biology ,Physical examination ,Clinodactyly ,Gene insertion ,Next generation sequencing ,Internal medicine ,medicine ,Disease severity ,Dietary intake ,Siblings ,Genomic dna ,Insulin resistance ,Follitropin ,lcsh:Genetics ,Young adult ,Clinical feature ,Preschool child ,Gemfibrozil ,novel mutation - Abstract
Background: Congenital leptin deficiency is a recessive genetic disorder associated with severe early-onset obesity. It is caused by mutations in the leptin (LEP) gene, which encodes the protein product leptin. These mutations may cause nonsense-mediated mRNA decay, defective secretion or the phenomenon of biologically inactive leptin, but typically lead to an absence of circulating leptin, resulting in a rare type of monogenic extreme obesity with intense hyperphagia, and serious metabolic abnormalities. Methods: We present two severely obese sisters from Colombia, members of the same lineal consanguinity. Their serum leptin was measured by MicroELISA. DNA sequencing was performed on MiSeq equipment (Illumina) of a next-generation sequencing (NGS) panel involving genes related to severe obesity, including LEP. Results: Direct sequencing of the coding region of LEP gene in the sisters revealed a novel homozygous missense mutation in exon 3 [NM_002303.3], C350G>, T [p.C117F]. Detailed information and clinical measurements of these sisters were also collected. Their serum leptin levels were undetectable despite their markedly elevated fat mass. Conclusions: The mutation of LEP, absence of detectable leptin, and the severe obesity found in these sisters provide the first evidence of monogenic leptin deficiency reported in the continents of North and South America.
- Published
- 2019
12. Patents on Enhancing the Potency and Longevity of Highly Valuable Protein Hormones
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Avri Havron, Fuad Fares, and Eyal Fima
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Protein Denaturation ,endocrine system ,Peptide Hormones ,Recombinant Fusion Proteins ,Endocrinology, Diabetes and Metabolism ,Peptide ,Follitropin ,Pharmacology ,Biology ,Peptide hormone ,Human chorionic gonadotropin ,Patents as Topic ,Follicle-stimulating hormone ,In vivo ,Drug Discovery ,medicine ,Animals ,Humans ,Immunology and Allergy ,Erythropoietin ,chemistry.chemical_classification ,Human Growth Hormone ,Protein Stability ,Biochemistry (medical) ,General Medicine ,Clinical trial ,chemistry ,Drug Design ,Follicle Stimulating Hormone, Human ,Half-Life ,medicine.drug - Abstract
One major issue regarding the clinical use of many peptides is their short half-life span in the body, due to the rapid clearance from the circulation. Thus, at the clinical level, there is a need for a regime of frequent injections of the peptides into the patients to overcome this low stability factor. The major strategies for overcoming this problem by pharmaceutical companies are based on chemical techniques and using specific peptidase inhibitors or cocktails. For this purpose, the cassette gene contains the sequence of the carboxyl-terminal peptide (CTP) of human chorionic gonadotropin β subunit which was ligated to the coding sequence of follitropin (FSH), erythropoietin (EPO), growth hormone (GH) and thus to increase the longevity and bioactivity of these proteins in vivo. Interestingly, FSH-CTP and GH-CTP were found to be not immunogenic in humans. FSH-CTP was approved by The European Commission. In addition, GH-CTP is not toxic and it passed successfully clinical trials Phase II in adults. Thus, using this technology seems to be promising in designing long acting peptides. Development of long acting peptides will diminish the cost of these drugs and perhaps reduce the number of injections in the clinical protocols. The article also summarizes some relevant patents.
- Published
- 2013
13. Role of Cysteine Residues in the Carboxyl-Terminus of the Follicle-Stimulating Hormone Receptor in Intracellular Traffic and Postendocytic Processing
- Author
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Teresa Zariñán, Alfredo Ulloa-Aguirre, Brenda Melo-Nava, Marco Allán Pérez-Solis, Eduardo Jardón-Valadez, Jean A. Castillo-Badillo, Arturo Aguilar-Rojas, Nathalie Gallay, Patricia Casas-González, Eric Reiter, José L. Maravillas-Montero, Research Unit in Reproductive Medicine, Instituto Mexicano del Seguro Social [Mexico City, Mexico] (IMSS), Research Support Network, Universidad National Autonoma de Mexico-Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán - National Institute of Medical Science and Nutrition Salvador Zubiran [Mexico], Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán - National Institute of Medical Science and Nutrition Salvador Zubiran [Mexico], Department of Earth Resources [Lerma), Universidad Autónoma Metropolitana, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Université Francois Rabelais [Tours], Biosynthesis Group (Institute of Molecular Biosciences), Instituto Mexicano del Seguro Social, Universidad National Autonoma de Mexico-Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán - National Institute of Medical Science and Nutrition Salvador Zubiran, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán - National Institute of Medical Science and Nutrition Salvador Zubiran, Department of Earth Resources, Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT] ,endocrine system ,follicle-stimulating hormone ,follicle-stimulating hormone receptor ,follitropin ,internalization ,palmitoylation ,recycling ,Endosome ,media_common.quotation_subject ,récepteur hormonal ,030209 endocrinology & metabolism ,Biology ,03 medical and health sciences ,symbols.namesake ,0302 clinical medicine ,Endocrinology ,Palmitoylation ,Lysosome ,medicine ,Internalization ,Receptor ,lcsh:QH301-705.5 ,hormonal receptor ,media_common ,Original Research ,Endoplasmic reticulum ,résidu cystéine ,Cell Biology ,Golgi apparatus ,030104 developmental biology ,medicine.anatomical_structure ,Biochemistry ,lcsh:Biology (General) ,symbols ,Follicle-stimulating hormone receptor ,follicule ,Developmental Biology ,Autre (Sciences du Vivant) - Abstract
Posttranslational modifications occurring during the biosynthesis of G protein-coupled receptors include glycosylation and palmitoylation at conserved cysteine residues located in the carboxyl-terminus (Ctail) of the receptor. In a number of these receptors, these modifications play an important role in receptor function and particularly, in intracellular trafficking. In the present study, the three cysteine residues present in the carboxyl-terminus of the human FSHR were replaced with glycine by site-directed mutagenesis. Wild-type and mutant (Cys627/629/655Gly) FSHRs were then transiently expressed in HEK-293 cells and analyzed for cell-surface plasma membrane expression, agonist-stimulated signaling and internalization, and postendocytic processing in the abscence and presence of lysosome and/or proteasome inhibitors. Compared with the wild-type FSHR, the triple mutant FSHR exhibited ~70% reduction in plasma membrane expression as well as a profound attenuation in agonist-stimulated cAMP production and ERK1/2 phosphorylation. Incubation of HEK-293 cells expressing the wild-type FSHR with 2-bromopalmitate (palmitoylation inhibitor) for 6 h, decreased plasma membrane expression of the receptor by ~30%. The internalization kinetics and β-arrestin 1 and 2 recruitment were similar between the wild-type and triple mutant FSHR as disclosed by assays performed in non-equilibrium binding conditions and by confocal microscopy. Cells expressing the mutant FSHR recycled the internalized FSHR back to the plasma membrane less efficiently than those expressing the wild-type FSHR, an effect that was counteracted by proteasome but not by lysosome inhibition. These results indicate that replacement of the cysteine residues present in the carboxyl-terminus of the FSHR, impairs receptor trafficking from the endoplasmic reticulum/Golgi apparatus to the plasma membrane and its recycling from endosomes back to the cell surface following agonist-induced internalization. Since in the FSHR these cysteine residues are S-palmitoylated, the data presented emphasize on this posttranslational modification as an important factor for both upward and downward trafficking of this receptor.
- Published
- 2016
14. Change in the ovarian environment after hysterectomy with bilateral salpingectomy: is it the technique or surgery itself?
- Author
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Kemal Özerkan, B. Cetinkaya Demir, Mehmet Aral Atalay, Uludağ Üniversitesi/Tıp Fakültesi/Kadın Hastalıkları ve Doğum Anabilim Dalı., Atalay, Mehmet Aral, Demir, Bilge Çetinkaya, Özerkan, Kemal, AAH-9834-2021, and AAH-9791-2021
- Subjects
Inhibin ,Anti-Mullerian Hormone ,endocrine system diseases ,Physiology ,medicine.medical_treatment ,Laparoscopic surgery ,Sexual intercourse ,Gray scale echography ,Bilateral Salpingectomy ,Uterus weight ,Follicle-stimulating hormone ,0302 clinical medicine ,Salpingectomy ,Fsh ,Obstetrics & gynecology ,Postoperative Period ,Reserve ,Priority journal ,Uterine Diseases ,030219 obstetrics & reproductive medicine ,biology ,Estradiol ,Bilateral salpingectomy ,Obstetrics and Gynecology ,Anti-Müllerian hormone ,Organ Size ,Middle Aged ,Preservation ,female genital diseases and pregnancy complications ,Estradiol blood level ,Blood ,medicine.anatomical_structure ,Ovarian Neoplasms ,Fallopian Tubes ,Carcinoma In Situ ,030220 oncology & carcinogenesis ,Level ,Diagnostic imaging ,Inhibin B ,Uterus disease ,Female ,Menopause ,Longitudinal study ,Luteinizing hormone blood level ,Luteinizing hormone ,Human ,Laparoscopic hysterectomy ,Adult ,endocrine system ,medicine.medical_specialty ,Ovary function ,Reproductive biology ,Ovary ,Major clinical study ,Hysterectomy ,Article ,03 medical and health sciences ,Total abdominal hysterectomy ,medicine ,Humans ,Women ,Inhibins ,Prospective study ,Follitropin blood level ,Gynecology ,Abdominal hysterectomy ,business.industry ,Follitropin ,Luteinizing Hormone ,Ovarian function ,Surgery ,Outcome assessment ,Reproductive Medicine ,biology.protein ,Benign uterus tumor ,Follicle Stimulating Hormone ,business ,Muellerian inhibiting factor ,Hormone - Abstract
Objective: To compare the effects of total laparoscopic hysterectomy with bilateral salpingectomy (TLH-BS) and total abdominal hysterectomy with bilateral salpingectomy (TAH-BS) on ovarian function among women of reproductive age. Study design: One hundred and three patients with a diagnosis of benign uterine disorder were divided into two groups in this prospective longitudinal study. Patients who had never had sexual intercourse and patients with uterovaginal disproportion underwent TAH-BS (n = 57), and the remaining patients (n = 46) underwent TLH-BS. Ovarian function was assessed before and 6 months after surgery; ovarian volume was assessed by gray-scale ultrasonography, and levels of serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), oestradiol (E2), anti-Mullerian hormone (AMH) and inhibin B were measured. Results: Postoperative serum FSH, LH and inhibin B decreased significantly in both groups. Postoperative serum E2 did not change significantly. Postoperative serum AMH and ovarian volume decreased significantly in the TAH-BS group (p = 0.016 and p < 0.001, respectively), but not in the TLH-BS group. Significant differences were observed between the TLH-BS and TAH-BS groups with respect to change in FSH (p = 0.012) and ovarian volume (p = 0.001); between-group differences were not significant for changes in AMH and inhibin B. Conclusions: Although serum AMH did not change significantly in patients who underwent TLH-BS, ovarian aging commenced following both surgical procedures.
- Published
- 2016
15. SHBG levels correlate with insulin resistance in postmenopausal women
- Author
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Mehmet Bastemir, Fulya Akin, Bunyamin Kaptanoglu, and Esma Alkis
- Subjects
Overweight ,Sex hormone-binding globulin ,Risk Factors ,insulin resistance ,Sex Hormone-Binding Globulin ,Hyperinsulinemia ,Sex hormones ,anthropometry ,resistance ,biology ,Metabolic Syndrome X ,article ,Middle Aged ,sex hormone binding globulin ,waist circumference ,Metabolic syndrome ,Postmenopause ,Postprandial ,hyperinsulinemia ,luteinizing hormone ,Biological Markers ,Female ,medicine.symptom ,Adult ,Aged ,Biomarkers/*blood ,Glucose Intolerance/blood/epidemiology ,Humans ,Hyperinsulinism/*blood/epidemiology ,Insulin Resistance ,Metabolic Syndrome/blood/epidemiology ,Obesity/blood/epidemiology ,Postmenopause/*blood ,Prediabetic State/blood/epidemiology ,Predictive Value of Tests ,Premenopause/blood ,Sensitivity and Specificity ,Sex Hormone-Binding Globulin/*metabolism ,follitropin ,hormones, hormone substitutes, and hormone antagonists ,medicine.medical_specialty ,premenopause ,hormone blood level ,sex hormone ,Prediabetic State ,Insulin resistance ,Hyperinsulinism ,Internal medicine ,Glucose Intolerance ,Internal Medicine ,medicine ,controlled study ,Obesity ,human ,business.industry ,medicine.disease ,thyroid hormone ,major clinical study ,Endocrinology ,Sex hormone binding globulin ,Insulin ,biology.protein ,business ,Biomarkers - Abstract
Background: Overweight or central obesity is generally associated with increases in fasting insulin levels, insulin resistance, and glucose intolerance and has been identified as a target for new therapeutic strategies, including early change in lifestyle. Early biochemical markers for identifying at-risk patients will be useful for prevention studies. The aim of this study is to investigate whether or not SHBG level is a useful index of hyperinsulinemia and/or insulin resistance in pre- and postmenopausal obese women. At the same time, the relationship between SHBG concentrations and features of the metabolic syndrome were evaluated. Methods: 229 women were eligible for this study. MetS was defined by using a modification of the ATP III guidelines. All patients were euthyroid, obese and overweight, 25 to 69 years of age. Subjects were divided into groups of premenopausal women (n = 125) and postmenopausal women (n = 104). Various fatness and fat distribution parameters, SHBG, sex hormones, FSH, LH, thyroid hormones, serum levels of fasting and postprandial glucose, lipid profile, uric acid and serum insulin, and blood pressure were measured. Results: No significant difference was found in mean SHBG levels between pre- and postmenopausal obese women in this study (p = 0.866). In premenopausal obese women, SHBG correlated negatively with BMI, waist circumference, fasting glucose, uric acid levels and FAI. In postmenopausal obese women, SHBG correlated negatively with fasting glucose, postprandial plasma glucose, fasting insulin, HOMA-IR and FAI and positively with HDL. SHBG had a significant inverse association with MetS parameters only in postmenopausal women, also after adjusting for BMI, age and estradiol. Conclusions: Obesity may influence the levels of endogenous sex steroid, especially after menopause. SHBG concentrations are correlated with features of the metabolic syndrome, particularly in postmenopausal obese women. These results suggest that SHBG may be an index of insulin resistance in postmenopausal obese women. © 2008 European Federation of Internal Medicine.
- Published
- 2009
16. STUDIES ON PITUITARY FOLLITROPIN
- Author
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M.R. Sairam and Choh Hao Lp
- Subjects
endocrine system ,biology ,Chemistry ,Protein subunit ,Alpha (ethology) ,Follitropin ,Biological activity ,Biochemistry ,Molecular biology ,Aspartic acid ,biology.protein ,Protein quaternary structure ,ATP synthase alpha/beta subunits ,G alpha subunit - Abstract
The alpha and beta subunits of human follitropin were isolated in a high state of purity. The tryptophan fluorescence of the native hormone and the isolated beta subunit are different. The N-terminus of the alpha and beta subunits was identified as valine and aspartic acid respectively. While recombination of the isolated alpha and beta subunits restores the electrophoretic mobility of the intact hormone, its receptor binding activity cannot be fully regenerated. Substitution of the human follitropin alpha by an ovine lutropin alpha subunit, to form a recombinant with the follitropin beta subunit, generates a complex with 2-3 receptor binding activity of the native human follitropin and the same activity as ovine follitropin. Acylation of the intact hormone does not disrupt the quaternary structure but leads to complete inactivation. Acylation studies with the subunits suggests the crucial role of the epsilon-amino groups of the alpha subunit in determining biological activity.
- Published
- 2009
17. Follicular dynamics and plasma FSH and progesterone concentrations during follicular deviation in the first post-ovulatory wave in Nelore (Bos indicus) heifers
- Author
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Andrea Renesto, Guilherme de Paula Nogueira, Leonardo F.C. Brito, Caliê Castilho, and Joaquim Mansano Garcia
- Subjects
Ovulation ,medicine.medical_specialty ,media_common.quotation_subject ,Follitropin ,Biology ,Andrology ,Follicle-stimulating hormone ,Follicle ,Endocrinology ,Ovarian Follicle ,Food Animals ,Internal medicine ,Follicular phase ,medicine ,Animals ,Ovarian follicle ,Progesterone ,media_common ,Estrous cycle ,General Medicine ,medicine.anatomical_structure ,Cattle ,Female ,Animal Science and Zoology ,Follicle Stimulating Hormone ,Hormone - Abstract
The objective of the present study was to characterize ovarian follicular dynamics and hormone concentrations during follicular deviation in the first wave after ovulation in Nelore (Bos indicus) heifers. Ultrasonographic exams were performed and blood samples were collected every 12h from the day of estrus until 120-144 h after ovulation in seven females. Deviation was defined as the point at which the growth rate of the dominant follicle became greater than the growth rate of the largest subordinate follicle. Deviation occurred approximately 65 h after ovulation. Growth rate of the dominant follicle increased (P0.05) after deviation, while growth rate of the subordinate follicle decreased (P0.05). Diameter of the dominant follicle did not differ from the subordinate follicle at deviation (approximately 5.4mm). The dominant follicle (7.6mm) was larger (P0.05) than the subordinate follicle (5.3mm) 96 h after ovulation or 24h after deviation. Plasma FSH concentrations did not change significantly during the post-ovulatory period. The first significant increase in mean plasma progesterone concentration occurred on the day of follicular deviation. In conclusion, the interval from ovulation to follicular deviation (2.7 days) was similar to that previously reported in B. taurus females, but follicles were smaller. Diameters of the dominant follicle and subordinate follicle did not differ before deviation and deviation was characterized by an increase in dominant follicle and decrease in subordinate follicle growth rate. Variations in FSH concentrations within 12-h intervals were not involved in follicular deviation in Nelore heifers.
- Published
- 2007
18. Follitropin receptors contain cryptic ligand binding sites
- Author
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John E. Kerrigan, Rebecca V. Myers, Win Lin, Michael P. Bernard, Donghui Cao, and William R. Moyle
- Subjects
endocrine system ,medicine.medical_specialty ,COS cells ,Chinese hamster ovary cell ,Follitropin ,Biology ,biology.organism_classification ,Biochemistry ,Cell biology ,Transmembrane domain ,Endocrinology ,Internal medicine ,medicine ,Cricetulus ,Binding site ,Receptor ,Molecular Biology ,Peptide sequence ,hormones, hormone substitutes, and hormone antagonists - Abstract
Human choriogonadotropin (hCG) and follitropin (hFSH) have been shown to contact different regions of the extracellular domains of G-protein coupled lutropin (LHR) and follitropin (FSHR) receptors. We report here that hCG and hFSH analogs interact with different regions of an FSHR/LHR chimera having only two unique LHR residues and that binds both hormones with high affinity. hCG and hFSH analogs dock with this receptor chimera in a manner similar to that in which they bind LHR and FSHR, respectively. This shows that although the FSHR does not normally bind hCG, it contains a cryptic lutropin binding site that has the potential to recognize hCG in a manner similar to the LHR. The presence of this cryptic site may explain why equine lutropins bind many mammalian FSHR and why mutations in the transmembrane domain distant from the extracellular domain enable the FSHR to bind hCG. The leucine-rich repeat domain (LRD) of the FSHR also appears to contain a cryptic FSH binding site that is obscured by other parts of the extracellular domain. This will explain why contacts seen in crystals of hFSH complexed with an LRD fragment of the human FSHR are hard to reconcile with the abilities of FSH analogs to interact with membrane G-protein coupled FSHR. We speculate that cryptic lutropin binding sites in the FSHR, which are also likely to be present in thyrotropin receptors (TSHR), permit the physiological regulation of ligand binding specificity. Cryptic FSH binding sites in the LRD may enable alternate spliced forms of the FSHR to interact with FSH.
- Published
- 2007
19. Single-Molecule Analyses of Fully Functional Fluorescent Protein-Tagged Follitropin Receptor Reveal Homodimerization and Specific Heterodimerization with Lutropin Receptor1
- Author
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James A. Dias, Joseph E. Mazurkiewicz, Alfredo Ulloa-Aguirre, Katharine Herrick-Davis, Barbara Lindau-Shepard, Margarida Barroso, and Richard M. Thomas
- Subjects
Granulosa cell differentiation ,endocrine system ,medicine.medical_specialty ,HEK 293 cells ,Fluorescence correlation spectroscopy ,Follitropin ,Cell Biology ,General Medicine ,Biology ,Fusion protein ,Cell biology ,Cell membrane ,Endocrinology ,Förster resonance energy transfer ,medicine.anatomical_structure ,Reproductive Medicine ,Internal medicine ,medicine ,Receptor - Abstract
We have previously shown that the carboxyl terminus (cT) of human follicle-stimulating hormone (FSH, follitropin) receptor (FSHR) is clipped before insertion into the plasma membrane. Surprisingly, several different constructs of FSHR fluorescent fusion proteins (FSHR-FPs) failed to traffic to the plasma membrane. Subsequently, we discovered that substituting the extreme cT of luteinizing hormone (LH) receptor (LHR) to create an FSHR-LHRcT chimera has no effect on FSHR functionality. Therefore, we used this approach to create an FSHR-LHRcT-FP fusion. We found this chimeric FSHR-LHRcT-FP was expressed in HEK293 cells at levels similar to reported values for FSHR in human granulosa cells, bound FSH with high affinity, and transduced FSH binding to produce cAMP. Quantitative fluorescence resonance energy transfer (FRET) analysis of FSHR-LHRcT-YFP/FSHR-LHRcT-mCherry pairs revealed an average FRET efficiency of 12.9 ± 5.7. Advanced methods in single-molecule analyses were applied in order to ascertain the oligomerization state of the FSHR-LHRcT. Fluorescence correlation spectroscopy coupled with photon-counting histogram analyses demonstrated that the FSHR-LHRcT-FP fusion protein exists as a freely diffusing homodimer in the plasma membrane. A central question is whether LHR could oligomerize with FSHR, because both receptors are coexpressed in differentiated granulosa cells. Indeed, FRET analysis revealed an average FRET efficiency of 14.4 ± 7.5 when the FSHR-LHR cT-mCherry was coexpressed with LHR-YFP. In contrast, coexpression of a 5-HT2cVSV-YFP with FSHR-LHR cT-mCherry showed only 5.6 ± 3.2 average FRET efficiency, a value indistinguishable from the detection limit using intensity-based FRET methods. These data demonstrate that coexpression of FSHR and LHR can lead to heterodimerization, and we hypothesize that it is possible for this to occur during granulosa cell differentiation.
- Published
- 2015
20. Controversial role of inhibin α-subunit gene in the aetiology of premature ovarian failure
- Author
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Eduardo H. Charreau, Victoria Sundblad, Liliana Dain, Stella Campo, Luz Andreone, and Violeta A. Chiauzzi
- Subjects
Candidate gene ,endocrine system diseases ,genotype ,inhibin A ,Primary Ovarian Insufficiency ,inhibin B ,Cohort Studies ,Loss of heterozygosity ,Gene Frequency ,Polymorphism (computer science) ,Genotype ,cytosine ,adult ,Rehabilitation ,allele ,article ,risk assessment ,Obstetrics and Gynecology ,alpha chain ,Inhibin α-subunit gene ,female genital diseases and pregnancy complications ,Premature ovarian failure ,female ,follicular phase ,nucleic acid base substitution ,Female ,Amenorrhea ,medicine.symptom ,follitropin ,Risk ,Heterozygote ,endocrine system ,medicine.medical_specialty ,Argentina ,premature ovarian failure ,Biology ,amenorrhea ,statistical analysis ,Internal medicine ,thymine ,medicine ,Humans ,inhibin ,heterozygosity ,Inhibins ,controlled study ,human ,Allele ,Polymorphism, Genetic ,Inhibin levels ,negative feedback ,Odds ratio ,medicine.disease ,major clinical study ,Endocrinology ,confidence interval ,Reproductive Medicine ,DNA polymorphism - Abstract
Background: Premature ovarian failure (POF) is characterized by hypergonadotropic amenorrhoea before the age of 40. Inhibin α-subunit (INHα) gene is proposed as a candidate gene due to its role in negative feedback control of FSH. Methods: Polymorphism -16 C>T of INHα gene was studied in 61 POF patients and 82 controls above 40 years old (C > 40). Substitution 769G>A was studied in 59 POF patients, 76 C > 40 and 73 controls below 40 years old (C < 40). Results: No significant difference in risk of POF development for -16T allele was found when comparing idiopathic POF (I-POF) with C > 40 (Odds ratio = 1.46; 95% confidence interval = 0.63-3.19). Implication of -16C>T polymorphism in serum inhibin levels was analysed in 46 controls, and no significant differences (P > 0.05) were found between CC and CT + TT genotype groups when comparing either mid-follicular phase Pro-αC and inhibin B values or mid-luteal phase Pro-αC and inhibin A values. Heterozygosity for substitution 769G>A was found in 1 of 59 POF woman, 2 of 76 C > 40 and 6 of 73 C < 40. Presence of this su bstitution in a relevant number of control subjects is herein described for the first time. Conclusion: Our results indicate that -16C>T and 769G>A variants in INHα gene may not be associated to POF disease. © 2006 Oxford University Press. Fil:Sundblad, V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Charreau, E.H. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.
- Published
- 2006
21. Serum HLA-G levels in women with polycystic ovary syndrome
- Author
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Ozer Oztekin, Yasar Enli, Semin Fenkci, Ümit Çabuş, and Veysel Fenkci
- Subjects
leukocyte count ,endocrine system diseases ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,cholesterol blood level ,insulin blood level ,Body Mass Index ,hyperandrogenism ,Follicle-stimulating hormone ,Endocrinology ,Sex hormone-binding globulin ,androgen blood level ,gonadotropin blood level ,high density lipoprotein cholesterol ,follitropin blood level ,insulin resistance ,Sex Hormone-Binding Globulin ,PCOS ,Medicine ,oxidative stress ,Testosterone ,glucose ,glutathione ,clinical article ,biology ,adult ,malonaldehyde ,Obstetrics and Gynecology ,sex hormone binding globulin ,testosterone blood level ,Polycystic ovary ,Lipids ,female ,HLA G antigen ,priority journal ,luteinizing hormone ,young adult ,Female ,Insulin resistance ,ovary ,Luteinizing hormone ,follitropin ,Polycystic Ovary Syndrome ,Adult ,medicine.medical_specialty ,insulin ,Dyslipidemias/blood/complications ,Follicle Stimulating Hormone/blood ,HLA-G Antigens/*blood ,Humans ,Hyperandrogenism/blood/complications ,Insulin Resistance/*physiology ,Lipids/blood ,Luteinizing Hormone/blood ,Oxidative Stress/physiology ,Polycystic Ovary Syndrome/*blood/complications ,Sex Hormone-Binding Globulin/metabolism ,Testosterone/blood ,Young Adult ,ovarian hyperandrogenism ,complication ,androgen ,Article ,lipid ,blood ,Internal medicine ,ovary polycystic disease ,controlled study ,human ,gonadotropin ,Dyslipidemias ,HLA-G Antigens ,business.industry ,Free androgen index ,Insulin ,Hyperandrogenism ,Ovary ,disease association ,dyslipidemia ,Luteinizing Hormone ,medicine.disease ,body mass ,disease assessment ,Oxidative Stress ,glucose blood level ,homeostasis model assessment of insulin resistance ,protein blood level ,physiology ,biology.protein ,luteinizing hormone blood level ,Follicle Stimulating Hormone ,Insulin Resistance ,business ,metabolism - Abstract
This study was designed to determine serum human leukocyte antigen-G (HLA-G) levels and establish whether serum HLA-G level is related with insulin resistance, oxidative stress, dyslipidemia and ovarian hyperandrogenism in women with polycystic ovary syndrome (PCOS). Twenty-five patients with PCOS and 23 healthy control women were evaluated in this study. Serum HLA-G, lipid fractions, glucose, insulin, malondialdehyde (MDA), glutathione (GSH), white blood cell (WBC), sex hormone-binding globulin (SHBG) and other hormone (gonadotropins and androgens) levels were measured. The estimate of insulin resistance was calculated by homeostasis model assessment (HOMA-IR). Serum luteinizing hormone (LH), total testosterone, fasting insulin, WBC levels and LH/follicle-stimulating hormone (FSH) ratio, free androgen index (FAI) and HOMA-IR values were significantly higher in patients with PCOS compared with healthy women. However, the women with PCOS had considerably lower serum FSH, SHBG, MDA, GSH and HLA-G levels than healthy subjects. HLA-G was inversely related with HOMA-IR, FAI, LH/FSH ratio and WBC, but positively with high-density lipoprotein cholesterol. Decreased serum HLA-G level may be related with insulin resistance, ovarian hyperandrogenism and oxidative stress in women with PCOS. Nevertheless, the exact role of HLA-G in the pathogenesis of the disease remains to be elucidated. © 2014 Informa UK Ltd.
- Published
- 2014
22. Effect of season and gonadotropins on the superovulatory response in camel (Camelus dromedarius)
- Author
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Manzoor A. Nowshari and Syed Azmal Ali
- Subjects
medicine.medical_specialty ,Camelus ,media_common.quotation_subject ,Superovulation ,Follitropin ,Ovary ,Breeding ,Biology ,Chorionic Gonadotropin ,Gonadotropin preparations ,Follicle ,Animal science ,Ovarian Follicle ,Food Animals ,Internal medicine ,Follicular phase ,medicine ,Animals ,Small Animals ,Ovulation ,Progesterone ,Ultrasonography ,media_common ,Equine ,Embryo ,Embryo Transfer ,Endocrinology ,medicine.anatomical_structure ,Tissue and Organ Harvesting ,Female ,Animal Science and Zoology ,Seasons ,Follicle Stimulating Hormone ,Estrus Synchronization ,Vetrepharm - Abstract
The purpose of the present investigation was to study the extent to which season and the gonadotropin preparation interferes with the superovulatory response in the dromedary. Adult camels were treated for superovulation during the breeding (November to April) and non-breeding season (May to October). Animals were synchronized by daily i.m. injections of progesterone (125 mg/animal/day, Jurox, UK) for 10 to 14 days. Superovulation was induced by 400mg pFSH alone (Follitropin V, Vetrepharm, Canada) administered in eight descending doses at 12h intervals or a combination of PMSG (2000IU, Folligon, Intervet, The Netherlands), injected with last injection of progesterone and 400mg pFSH in eight descending doses. The follicular development was daily assessed by ultrasonography of the ovaries. The donors were classified as per their response to the superovulatory treatment into very good (10 follicles), good (5-10 follicle), poor (2-4 follicles) or no response (1 or no follicle) on each ovary. Ovulation was induced by injecting 3000 IU hCG (Chorulon, Intervet) at the time of first mating. The donors were mated twice at an interval of 12h when all or most of the follicles reached to a size of about 1.0-1.7 cm. Camels were flushed non-surgically on Day 6 or 7 after the ovulation. The proportion of camels showing very good response during the breeding as well as non-breeding season was higher (P0.05) when a combination of pFSH and eCG was used compared with pFSH only. There was no difference (P0.05) in the proportion of donors flushed successfully (embryos recovered) when treated either with a combination of pFSH and eCG or pFSH alone during the breeding and non-breeding season. The rate of recovery of ova/embryos and proportion of transferable embryos was higher (P0.05) when donors were treated with pFSH+eCG compared with pFSH only during the breeding as well as non-breeding season. The results may indicate that ova/embryo recovery rate of the dromedary is influenced by the gonadotropin preparation but is not appreciably affected by the season.
- Published
- 2005
23. Role of follitropin receptor signaling in nuclear protein transitions and chromatin condensation during spermatogenesis
- Author
-
M. Ram Sairam, Weirong Xing, and Hanumanthappa Krishnamurthy
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,Chromosomal Proteins, Non-Histone ,Biophysics ,Follitropin ,Biology ,Biochemistry ,Chromatin remodeling ,Mice ,Prophase ,Internal medicine ,Testis ,medicine ,Animals ,Spermatogenesis ,Molecular Biology ,Cells, Cultured ,Nuclear Proteins ,Cell Biology ,Chromatin Assembly and Disassembly ,Sertoli cell ,Spermatozoa ,Sperm ,Chromatin ,Cell biology ,Endocrinology ,medicine.anatomical_structure ,Receptors, FSH ,Follicle Stimulating Hormone ,Follicle-stimulating hormone receptor ,Signal Transduction - Abstract
Follitropin receptor (FSHR) in testicular Sertoli cells mediates signaling by pituitary follitropin (FSH) promoting intercellular communication with germ cells for normal spermatogenesis. Using receptor knockout mice we examined changes in sperm nucleoproteins and chromatin architecture. The expressions of transition proteins 1/2 (TP1/2) and protamine-2 (PRM-2) were greatly diminished at 21 days, but returned to normal at 35 days and 3 months after birth. However, protein components in chromatin were quite different. Western blots detected a reduction in PRM1/2 and prolonged retention of mono-ubiquitinated histone 2A (uH2A) in the epididymal sperm from adult mutants. Two forms of mono- and poly-uH2A were present in sonication-resistant testicular spermatids in normal mice, whereas only an elevated mono-uH2A was detectable in mutants. Decrease in PRM1/2 and retention of mono-uH2A was coincident with reduction in TP1/2 in premature spermatids. Thus lack of FSHR signaling impairs expression of TP1/2 and PRM-2 at an early stage of post-natal development causing delayed spermatogenesis. In the adult, absence of FSHR signaling prolongs retention of mono-uH2A, leading to impair transition of basic nucleoproteins and chromatin remodeling during mouse spermatogenesis.
- Published
- 2003
24. Follitropin-α versus human menopausal gonadotropin in an in vitro fertilization program
- Author
-
James M. Goldfarb and Nina Desai
- Subjects
Gynecology ,endocrine system ,medicine.medical_specialty ,Pregnancy ,In vitro fertilisation ,medicine.drug_class ,medicine.medical_treatment ,Obstetrics and Gynecology ,Follitropin ,Biology ,medicine.disease ,Embryo transfer ,Andrology ,Follicle-stimulating hormone ,Pregnancy rate ,Reproductive Medicine ,medicine ,Gestation ,Gonadotropin ,hormones, hormone substitutes, and hormone antagonists - Abstract
Objective To compare the efficacy of recombinant FSH and urinary-derived hMG for ovarian stimulation during IVF. Design Retrospective analysis of data from IVF cycles conducted over 15 months. Setting University hospital IVF unit. Patient(s) Three hundred twenty-four women undergoing their first to sixth IVF cycle. Intervention(s) After pituitary down-regulation, patients received recombinant FSH or hMG, according to personal choice. After hCG administration, patients underwent oocyte retrieval, oocyte fertilization, and embryo transfer. Main outcome measure(s) Implantation rate and clinical ongoing pregnancy rate per oocyte retrieval. Result(s) Patients who chose recombinant FSH were slightly younger than those who chose hMG (34.1 vs. 35.1 years, respectively). Although more embryos were transferred in the hMG group (3.6 vs. 3.2), the ongoing pregnancy and implantation rates were significantly higher in the recombinant FSH group (ongoing pregnancy rate, 50.0% vs. 36.2%). Conclusion(s) Recombinant FSH is more effective than hMG for ovarian stimulation in IVF cycles. This increased efficacy, which is achieved with fewer ampoules, is likely to offset the higher acquisition costs of recombinant FSH.
- Published
- 2003
25. Structural Biology of Human Follitropin and Its Receptor
- Author
-
Patrick Van Roey and James A. Dias
- Subjects
Models, Molecular ,endocrine system ,Conformational change ,Glycosylation ,Protein Conformation ,Amino Acid Motifs ,Molecular Sequence Data ,Thyrotropin ,Follitropin ,Protein Sorting Signals ,Biology ,Chorionic Gonadotropin ,Protein Structure, Secondary ,Structure-Activity Relationship ,Humans ,Amino Acid Sequence ,Receptor ,Protein secondary structure ,chemistry.chemical_classification ,Sequence Homology, Amino Acid ,General Medicine ,Luteinizing Hormone ,Protein Structure, Tertiary ,Amino acid ,Biochemistry ,Structural biology ,chemistry ,Hormone receptor ,Mutagenesis, Site-Directed ,Receptors, FSH ,Follicle Stimulating Hormone ,Glycoprotein ,Dimerization ,Protein Binding - Abstract
In this review, the current understanding of structure-activity relationships of human follitropin and of the extracellular domain of its receptor is described. Comprehensive mutagenesis of human follitropin combined with the three-dimensional structure of human follitropin has ushered in a new era of understanding of how this complex hormone binds to and activates its receptor. Comparison of human choriogonadotropin and follitropin structures has proved invaluable in understanding how these human glycoprotein hormones have conserved primary sequence that enables high affinity binding while diverging in amino acids that provide specificity. Moreover, by comparison of the structures of deglycosylated and glycosylated human choriogonadotropin and glycosylated human follitropin, there appears to be no influence of oligosaccharides upon backbone conformation of human glycoprotein hormones. Extensive structure-activity relationships of human follitropin receptor have been studied, and new insights gained here as well. These studies indicate that follitropin binds to the central module of the extracellular domain of the follitropin receptor. Biophysical analyses of purified follitropin receptor extracellular domain further revealed conformational changes affected by hormone binding and by the solvent environment. Further, secondary structure analysis of the purified extracellular domain of follitropin receptor favors the leucine-rich repeat motif model of the glycoprotein hormone receptors. Together, the studies indicate that there are only a few residues that contribute to the overall energy of binding. Formation of a weak collisional complex between follitropin and its receptor likely involves complementation of compatible surfaces and steric hindrance by oligosaccharides, followed by conformational change and formation of active site residue salt bridges. In this regard and in light of these new data, current models of the glycoprotein hormone receptors may need to be re-evaluated.
- Published
- 2001
26. [Untitled]
- Author
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Jean-Noël Hugues, Isabelle Cedrin-Durnerin, Helene Bry-Gauillard, Bettina Bständig, and Michèle Uzan
- Subjects
Agonist ,endocrine system ,medicine.medical_specialty ,In vitro fertilisation ,medicine.drug_class ,medicine.medical_treatment ,Obstetrics and Gynecology ,Follitropin ,General Medicine ,Biology ,Triptorelin ,Embryo transfer ,Andrology ,Follicle-stimulating hormone ,Endocrinology ,Reproductive Medicine ,Internal medicine ,Gonadotropin-releasing hormone agonist ,Genetics ,medicine ,Gonadotropin ,hormones, hormone substitutes, and hormone antagonists ,Genetics (clinical) ,Developmental Biology ,medicine.drug - Abstract
Purpose: To compare the efficiency and efficacy of two recombinant human FSH (r-FSH) and urinary (u-FSH) preparations in patients undergoing superovulation for IVF-ET using a short-term gonadotropin releasing hormone agonist (GnRH-a) (Triptorelin) protocol. Methods: A total of 88 women undergoing IVF-ET were included in this prospective study. They were randomized to receive u-FSH (150 IU/d), follitropin-α (100 IU/d), or follitropin-β (100 IU/d) for 2 days, and dosages were subsequently adjusted according to the ovarian response. Results: The FSH dose required for the overall stimulation was significantly lower in patients treated with r-FSH than in those treated with u-FSH while serum FSH values were higher in the latter group. There were no statistically significant differences in ovarian response and IVF outcome between r-FSH preparations. Conclusions: Recombinant FSH preparations have a higher efficiency than urinary ones in patients undergoing IVF-ET using a short-term GnRH-a protocol. In this situation, the two recombinant follitropins have comparable effectiveness.
- Published
- 2001
27. Bifunctional hCG analogs adopt different conformations in LH and FSH receptor complexes
- Author
-
Michael P. Bernard, Yanhong Wang, and William R. Moyle
- Subjects
endocrine system ,Protein Conformation ,Recombinant Fusion Proteins ,Protein subunit ,Molecular Sequence Data ,Follitropin ,Chorionic Gonadotropin ,Biochemistry ,chemistry.chemical_compound ,Endocrinology ,Lutropin receptor ,Humans ,Amino Acid Sequence ,Bifunctional ,Receptor ,Molecular Biology ,Immunoassay ,Binding Sites ,biology ,Antibodies, Monoclonal ,Receptors, LH ,HCG - Human chorionic gonadotropin ,chemistry ,biology.protein ,Receptors, FSH ,Follicle Stimulating Hormone ,Antibody ,Follicle-stimulating hormone receptor ,Sequence Alignment ,Epitope Mapping ,hormones, hormone substitutes, and hormone antagonists ,Protein Binding - Abstract
Human reproduction requires specific interactions between follitropin (hFSH) and its receptor (FSHR) and between lutropin (hLH) or choriogonadotropin (hCG) and the lutropin receptor (LHR). Substitution of hFSH residues between hCG β-subunit cysteines 11–12 creates a bifunctional analog that binds both receptors. To understand the basis of this observation, we used antibody probes to compare the conformations of bifunctional analogs before and after they were complexed with each receptor. Introduction of hFSH residues between cysteines 11–12 changed a distant conformation-sensitive region created by the juxtaposition of the subunit aminotermini. This region, found not to contact either receptor, was altered further when bifunctional ligands bound FSHR. All other surfaces, detected in LHR complexes, were also recognized in FSHR complexes, an indication that bifunctional ligands bind both receptors in similar orientations. These observations suggest that unlike hCG or hFSH, bifunctional gonadotropins can acquire ‘lutropin’ and ‘follitropin’ conformations, a phenomenon accentuated by receptor contacts.
- Published
- 2000
28. Multiple Distant Amino Acid Residues Present in the Serpentine Region of the Follitropin Receptor Modulate the Rate of Agonist-induced Internalization
- Author
-
Mario Ascoli and Hiroshi Kishi
- Subjects
Agonist ,DNA, Complementary ,Arrestins ,medicine.drug_class ,Recombinant Fusion Proteins ,media_common.quotation_subject ,Molecular Sequence Data ,Follitropin ,Follitropin Receptor ,Biology ,Transfection ,Biochemistry ,Protein Structure, Secondary ,Cell Line ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Amino Acids ,Cloning, Molecular ,Receptor ,Internalization ,Molecular Biology ,media_common ,chemistry.chemical_classification ,Sequence Homology, Amino Acid ,Cell Membrane ,Cell Biology ,Molecular biology ,Recombinant Proteins ,Protein Structure, Tertiary ,Rats ,Amino acid ,Kinetics ,Transmembrane domain ,chemistry ,Receptors, FSH ,Intracellular ,Plasmids ,Protein Binding - Abstract
The amino acid sequences of the human (h) and rat (r) follitropin receptors (FSHR) are approximately 89% identical, but the half-time of internalization of agonist mediated by the rFSHR is approximately 3 times faster than that of the hFSHR. Chimeras of the hFSHR and the rFSHR showed that this difference in rate is dictated mostly by the serpentine domain. Further analysis identified six residues, two non-contiguous residues in the transmembrane helix 4 (Leu/Thr in the rFSHR and Met/Ile in the hFSHR), three non-contiguous residues in the third intracellular loop (Thr/Thr/Lys in the rFSHR and Ile/Asn/Arg in the hFSHR), and one in transmembrane helix 7 (Tyr in the rFSHR and His in the hFSHR) that are fully responsible for the difference in the rates of internalization of the hFSHR and the rFSHR.
- Published
- 2000
29. Bioassays of Gonadotropins
- Author
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Philippe Bouchard, Bart C.J.M. Fauser, Sophie Christin-Maitre, Claudine Vasseur, and Obstetrics & Gynecology
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,DNA, Complementary ,Granulosa cell ,Follitropin ,CHO Cells ,Biology ,Transfection ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Rats, Sprague-Dawley ,Radioligand Assay ,In vivo ,Cricetinae ,Internal medicine ,medicine ,Radioligand ,Animals ,Humans ,Bioassay ,Inhibins ,Testosterone ,Cloning, Molecular ,Luciferases ,Receptor ,Molecular Biology ,Immunoassay ,Granulosa Cells ,Sertoli Cells ,Leydig Cells ,Receptors, LH ,Sertoli cell ,Rats ,medicine.anatomical_structure ,Endocrinology ,Receptors, FSH ,Biological Assay ,Female ,Follicle-stimulating hormone receptor ,Gonadotropins ,hormones, hormone substitutes, and hormone antagonists - Abstract
Bioassays constitute a unique approach to determine the functional aspects of gonadotropins. Indeed, these highly complex glycoprotein hormones, including pituitary lutropin (LH) and follitropin (FSH), are heterogeneous in terms of both peptidic and carbohydrate moieties, and, as a consequence, the bioactivity of the different molecular forms often does not match their immunoreactivity. In this article, we review the different types of LH and FSH bioassays. Conventional methods for measuring FSH bioactivity are first described and include the in vivo Steelman and Pohley bioassay, the radioligand receptor assays (RRAs), the in vitro Sertoli cell bioassay, the in vitro granulosa cell bioassay, and the inhibin immunoassay. Recent methods based on cell lines transfected with cloned receptors, particularly the human FSH receptor, are then described. Methods for developing these assays are presented, and the advantages and disadvantages of the different bioassays are discussed.
- Published
- 2000
30. Synthesis of anα-(2,3)-Sialylated, Complex-Type Undecasaccharide
- Author
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Yukishige Ito, Joachim Seifert, and Matthias Lergenmüller
- Subjects
chemistry.chemical_classification ,Glycan ,Glycosylation ,biology ,Stereochemistry ,Regioselectivity ,Follitropin ,General Chemistry ,Complex type ,Catalysis ,chemistry.chemical_compound ,chemistry ,Biochemistry ,biology.protein ,Glycoprotein - Abstract
Onlytwobuildingblockswerenecessary to provide efficient access to the complex undecasaccharide 1 by means of two stereo- and regioselective glycosylation reactions. Stereocontrolled p-methoxybenzyl-assisted β-mannosylation afforded the critical β-mannoside linkage in 1, which is a constituent of the human glycoproteins follitropin and gonadotropin.
- Published
- 2000
31. Chicken lutropin acts like follitropin in rat ovarian follitropin receptor: An isoelectric focusing study
- Author
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Michiharu Kamiyoshi, Masa-aki Hattori, Yoshiko Fukuhara, Katsumi Wakabayashi, Atsushi Iwasawa, and Mitsuo Kawashima
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,animal structures ,Immunology ,Radioimmunoassay ,Follitropin ,Biology ,Iodine Radioisotopes ,Species Specificity ,Anterior pituitary ,Pituitary Gland, Anterior ,Internal medicine ,medicine ,Radioligand ,Animals ,Rats, Wistar ,Receptor ,Pharmacology ,Chromatography ,Isoelectric focusing ,Sepharose ,Ovary ,Luteinizing Hormone ,Rats ,Endocrinology ,medicine.anatomical_structure ,Isoelectric point ,Receptors, FSH ,Female ,Follicle Stimulating Hormone ,Isoelectric Focusing ,Follicle-stimulating hormone receptor ,Chickens ,hormones, hormone substitutes, and hormone antagonists - Abstract
This study investigates whether chicken lutropin (LH) specifically binds to rat ovarian follitropin (FSH) receptor and exerts FSH-like bioactivity. Glycoprotein fraction, prepared from the chicken anterior pituitary gland, was fractionated using isoelectric focusing within a pH range of 3.5-11. Analysis of the focused fractions, by a radioreceptor assay (RRA) specific for FSH in rats using rat ovarian homogenate as receptor source, and 125I-labeled rat FSH as radioligand, detected a large component having an isoelectric point of 10.25. This focusing profile obtained by RRA was quite similar to that obtained by a specific radioimmunoassay (RIA) for chicken LH, but clearly different from that obtained by a specific RIA for chicken FSH, indicating this RRA specifically recognizes chicken LH. Chicken LH fraction prepared from the electrofocused material was used for further studies. The chicken LH preparation was three times more potent than rat FSH in the RRA in displacing the radioligand bound to rat ovarian receptor, while chicken LH facilitated an 8-fold less production of estradiol in dispersed rat granulosa cells than rat FSH. These results suggest that chicken LH acts like rat FSH in rat ovarian FSH receptor, but receptor-binding activity is much higher than biological activity.
- Published
- 1998
32. Biotechnological Drugs for Reproductive Disorders
- Author
-
T. Strowitzki
- Subjects
Pharmacology ,endocrine system ,medicine.medical_specialty ,Urinary system ,Endogeny ,Follitropin ,Stimulation ,General Medicine ,Urofollitropin ,Biology ,Endocrinology ,Internal medicine ,Follicular phase ,medicine ,Pharmacology (medical) ,Menotropins ,hormones, hormone substitutes, and hormone antagonists ,Biotechnology ,medicine.drug ,Hormone - Abstract
Recombinant gonadotrophins, in particular recombinant follitropin (follicle-stimulating hormone; FSH), are now available for ovarian hyperstimulation for assisted reproduction. In contrast with urinary FSH (urofollitropin), follitropin is available in virtually unlimited quantities. Follitropin is as potent as urofollitropin in all protocols of ovarian stimulation. Furthermore, it shows an improved purity without contamination by urinary proteins not related to FSH, and can be injected subcutaneously by the patients themselves. In patients with complete luteinising hormone (LH) deficiency, follitropin stimulates follicular development, although serum levels of estradiol remain low. For this group of patients the addition of LH is necessary. Ongoing phase III studies on the use of recombinant LH in this indication will provide an answer to how much LH is needed in order to guarantee sufficient follicular growth and hormonal response. Gonadorelin (gonadotrophin releasing hormone; GnRH) analogues are used to avoid the surge of endogenous LH in ovarian stimulation protocols. Gonadorelin antagonists are now in clinical testing for the same indication. Gonadorelin antagonists allow sufficient suppression of endogenous LH levels. In contrast with gonadorelin analogues, they avoid any flare-up effect, i.e. an initial release of gonadotrophins from pituitary reservoirs. Recombinant gonadotrophins and gonadorelin antagonists are new tools towards a more individual approach to ovarian stimulation.
- Published
- 1997
33. Transforming growth factor-α stimulates insulin-like growth factor binding protein-4 (IGFBP-4) expression and blocks follicle-stimulating hormone regulation of IGFBP-4 production in rat granulosa cells
- Author
-
Francesc Piferrer, Shunichi Shimasaki, Danmei Li, and Gregory F. Erickson
- Subjects
TGF-α ,Granulosa cells ,medicine.medical_specialty ,TGF alpha ,medicine.medical_treatment ,Biology ,Biochemistry ,Insulin-like growth factor-binding protein ,Rats, Sprague-Dawley ,Follicle-stimulating hormone ,Endocrinology ,Internal medicine ,medicine ,Animals ,Pregnancy-Associated Plasma Protein-A ,IGFBP ,RNA, Messenger ,Northern blot ,Molecular Biology ,Cells, Cultured ,Granulosa Cells ,Protease ,Growth factor ,IGFBP-4 Protease ,Metalloendopeptidases ,Transforming Growth Factor alpha ,Transforming growth factor alpha ,Follitropin ,Somatomedin binding protein 4 ,IGFBP-4 ,TGF ,Rats ,Hormone inhibition ,Insulin-Like Growth Factor Binding Protein 4 ,biology.protein ,Protein expression ,Female ,Steady state (chemistry) ,Follicle Stimulating Hormone ,hormones, hormone substitutes, and hormone antagonists ,Transforming growth factor - Abstract
9 pages, 7 figures The ability of TGF-α to regulate insulin-like growth factor binding protein-4 (IGFBP-4), was investigated. Primary cultures of rat granulosa cells (GC) were grown in serum-free medium with rat (r) TGF-α and/or rFSH, and secreted IGFBP-4 protein and its steady state mRNA levels were measured by Western immunoblotting and Northern blotting, respectively. Control (untreated) cells secreted IGFBP-4 spontaneously, and the levels were increased by rTGF-α in a dose- and time-dependent manner. rTGF-α abolished FSH-induced IGFBP-4 protease activity and suppressed FSH-dependent effects on IGFBP-4 production. IGFBP-4 mRNA levels were decreased and increased by FSH and TGF-α, respectively, and TGF-α blocked the FSH effects. These results demonstrate that TGF-α is a potent stimulator of IGFBP-4 expression in rat GC and can overcome the regulatory effects of FSH on IGFBP-4 production. The consequence of these TGF-α effects is a marked, sustained increase in the levels of IGFBP-4 in the microenvironment This work was supported by NICHD Grants HD-29008 and HD-30308. F.P. was supported by a NATO postdoctoral fellowship
- Published
- 1997
34. Serum fetuin-A levels, insulin resistance and oxidative stress in women with polycystic ovary syndrome
- Author
-
Ozer Oztekin, Semin Fenkci, Yasar Enli, and Veysel Fenkci
- Subjects
Blood Glucose ,endocrine system diseases ,alpha-2-HS-Glycoprotein ,Endocrinology, Diabetes and Metabolism ,medicine.medical_treatment ,insulin blood level ,low density lipoprotein cholesterol ,hyperandrogenism ,Follicle-stimulating hormone ,Endocrinology ,Sex hormone-binding globulin ,androgen blood level ,gonadotropin blood level ,high density lipoprotein cholesterol ,follitropin blood level ,insulin resistance ,Malondialdehyde ,Sex Hormone-Binding Globulin ,PCOS ,oxidative stress ,Insulin ,glucose ,glutathione ,fetuin A ,prasterone sulfate ,clinical article ,biology ,adult ,malonaldehyde ,article ,Obstetrics and Gynecology ,sex hormone binding globulin ,testosterone blood level ,waist circumference ,Polycystic ovary ,superoxide dismutase ,myeloperoxidase ,female ,priority journal ,luteinizing hormone ,Androgens ,Luteinizing hormone ,follitropin ,Polycystic Ovary Syndrome ,medicine.medical_specialty ,Adolescent ,ovarian hyperandrogenism ,androgen ,dependent variable ,Young Adult ,Insulin resistance ,Internal medicine ,ovary polycystic disease ,medicine ,Humans ,controlled study ,human ,gonadotropin ,Dyslipidemias ,Peroxidase ,business.industry ,Free androgen index ,Hyperandrogenism ,disease association ,dyslipidemia ,nutritional and metabolic diseases ,medicine.disease ,independent variable ,Fetuin-A ,glucose blood level ,protein blood level ,testosterone ,biology.protein ,luteinizing hormone blood level ,Follicle Stimulating Hormone ,business - Abstract
This study was designed to determine serum Fetuin-A levels and establish whether serum Fetuin-A level is related with insulin resistance, oxidative stress, ovarian hyperandrogenism and dyslipidemia in women with polycystic ovary syndrome (PCOS). Twenty-two patients with PCOS and twenty-one healthy control women were evaluated in this controlled clinical study. Serum Fetuin-A, lipid fractions, glucose, insulin, malondialdehyde (MDA), myeloperoxidase (MPO), glutathione (GSH), superoxide dismutase (SOD) and other hormone (gonadotropins, androgens) levels were measured. The estimate of insulin resistance was calculated by homeostasis model assessment (HOMA-R). The women with PCOS had significantly higher serum fasting glucose, insulin, luteinizing hormone (LH), MDA, Fetuin-A levels, and LH/follicle-stimulating hormone (FSH) ratio, free androgen index (FAI), HOMA-IR than healthy women. However, sex hormone-binding globulin (SHBG) and GSH levels were significantly lower in patients with PCOS compared with controls. Fetuin-A was positively correlated with insulin, HOMA-IR and FAI. Multiple regression analysis revealed that FAI was strong predictor of serum Fetuin-A level. Serum Fetuin-A level was related with insulin resistance and ovarian hyperandrogenism in women with PCOS. These results suggest that Fetuin-A may have a role in triggering the processes leading to insulin resistance and androgen excess in PCOS. © 2013 Informa UK Ltd.
- Published
- 2013
35. Trafficking of the follitropin receptor
- Author
-
James A. Dias, Ilpo Huhtaniemi, Alfredo Ulloa-Aguirre, Eric Reiter, George R. Bousfield, Instituto Nacional de Salud Publica [Mexique] (INSP), Studium Consortium for Research and Training in Reproductive Sciences (sCORTS), Department of Health and Department of Biomedical Sciences, State University of New York (SUNY)-Wadsworth Center, Department of Biological Sciences, Wichita State University, Institute of Reproductive and Developmental Biology (IRDB), Imperial College London, Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), CONACyT (Mexico), The National Institutes of Health (Bethesda, MD), The Wellcome Trust and Academy of Finland, Région Centre, INRA, ANR, Absent, National Institute of Public Health = Instituto Nacional de Salud Pública [Cuernavaca, Mexique] (INSP), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), P. Michael Conn, and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
endocrine system ,[SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT] ,Arrestins ,intracellular trafficking ,media_common.quotation_subject ,Blotting, Western ,030209 endocrinology & metabolism ,Follitropin ,Biology ,recycling ,Article ,follitropin receptor ,03 medical and health sciences ,0302 clinical medicine ,expression ,Fluorescence Resonance Energy Transfer ,Animals ,Humans ,Immunoprecipitation ,Phosphorylation ,Internalization ,Receptor ,beta-Arrestins ,030304 developmental biology ,G protein-coupled receptor ,media_common ,follicle stimulating hormone ,0303 health sciences ,dimerization ,Beta-Arrestins ,Endoplasmic reticulum ,Ligand (biochemistry) ,internalization ,Protein Transport ,Biochemistry ,Hormone receptor ,Mutation ,Receptors, FSH ,Electrophoresis, Polyacrylamide Gel ,Protein Multimerization ,follitropin - Abstract
Chapitre 2; The follitropin or follicle-stimulating hormone receptor (FSHR) belongs to a highly conserved subfamily of the G protein-coupled receptor (GPCR) superfamily and is mainly expressed in specific cells in the gonads. As any other GPCR, the newly synthesized FSHR has to be correctly folded and processed in order to traffic to the cell surface plasma membrane and interact with its cognate ligand. In this chapter, we describe in detail the conditions and procedures used to study outward trafficking of the FSHR from the endoplasmic reticulum to the plasma membrane. We also describe some methods to analyze phosphorylation, beta-arrestin recruitment, internalization, and recycling of this particular receptor, which have proved useful in our hands for dissecting its downward trafficking and fate following agonist stimulation.
- Published
- 2013
36. Daily Rhythmic and Non-Rhythmic Variations of Follitropin, Lutropin, Testosterone, and Sex-Hormone-Binding Globulin in Men
- Author
-
Xavier Fuentes-Arderiu and José Valero-Politi
- Subjects
Adult ,Male ,medicine.medical_specialty ,Globulin ,medicine.drug_class ,education ,Clinical Biochemistry ,Follitropin ,Sex hormone-binding globulin ,Reference Values ,Sex Hormone-Binding Globulin ,Internal medicine ,medicine ,Humans ,Testosterone ,Circadian rhythm ,Saliva ,biology ,Biochemistry (medical) ,General Medicine ,Venous blood ,Luteinizing Hormone ,Androgen ,Circadian Rhythm ,Endocrinology ,biology.protein ,Follicle Stimulating Hormone ,Gonadotropin - Abstract
The circadian rhythmic variations of the serum concentrations of follitropin, lutropin, sex-hormone-binding globulin and testosterone, the ratio between the serum concentrations of testosterone and sex-hormone-binding globulin, and the salivary concentration of testosterone were investigated in a group of 13 apparently healthy men. Venous blood and salivary specimens were collected at 4-h intervals over a 24-h period. The circadian rhythms were studied by using a periodic function resulting from the sum of two cosine functions with periods of 24 and 12 h. The serum concentrations of follitropin and lutropin showed no significant rhythmic variations. For the salivary concentration of testosterone and for the ratio between the serum concentrations of testosterone and sex-hormone-binding globulin, only the cosine function with a period of 24 h was significant. Serum concentrations of sex-hormone binding globulin and testosterone were significantly affected by 24- and 12-h rhythmic components. Of the quantities studied, the salivary concentration of testosterone showed the greatest daily rhythmic variation (28.8% of the mean estimated over rhythm).
- Published
- 1996
37. Assisted conception. I--General principles
- Author
-
Paula Rowell and Peter Braude
- Subjects
Male ,Clinical Review ,endocrine system ,Reproductive Techniques, Assisted ,medicine.medical_treatment ,media_common.quotation_subject ,Follitropin ,Fertilization in Vitro ,Biology ,Insemination ,Intracytoplasmic sperm injection ,Andrology ,Pregnancy ,medicine ,Humans ,Ovulation ,Infertility, Male ,Insemination, Artificial ,Fertilisation ,General Environmental Science ,media_common ,In vitro fertilisation ,Artificial insemination ,General Engineering ,General Medicine ,Sperm ,Gamete Intrafallopian Transfer ,General Earth and Planetary Sciences ,Female ,Infertility, Female - Abstract
Although many assisted conception technologies exist—and have a bewildering array of acronyms—their principal aim is similar. They all aim to bring sperm and the egg close to each other to promote the chances of fertilisation and, ultimately, achieve a pregnancy. View this table: Acronyms used in assisted conception The three main types of assisted conception are intrauterine insemination, in vitro fertilisation, and intracytoplasmic sperm injection. Intrauterine insemination —Prepared sperm are deposited in the uterus at a time when ovulation is likely or assisted In vitro fertilisation —Fertilisation is aided by mixing eggs and sperm in the laboratory Intracytoplasmic sperm injection —A single sperm is injected directly into the egg cytoplasm to achieve fertilisation. Each of these assisted conception techniques requires three procedures: pharmacological stimulation of the ovary to promote the production of more than one egg (superovulation); laboratory preparation of the semen sample to yield a highly motile, morphologically normal population of sperm for insemination or injection (sperm preparation); and techniques to aid the union of sperm and egg (assisted fertilisation). Multifollicular development can be achieved by using oral antioestrogens, such as clomifene citrate or tamoxifen. However, more often multifollicular development requires injected preparations containing the pituitary hormone, follicle stimulating hormone (FSH). FSH used to be obtained from extracts of urine collected from postmenopausal women, which were then purified to various degrees to remove contaminating proteins and luteinising hormone. The extracts provided a preparation of human menopausal gonadotrophins marketed as human menotropins—for example, Merional, Menogon, Menopur. Variation within batches of gonadotrophins, and the increasing unacceptability of injecting biologically derived substances, has led to the more widespread use of recombinant products. These include Gonal-F (follitropin α) and Puregon (follitropin β). Although chemically pure, and …
- Published
- 2003
38. Comparison of aneuploidy frequencies between in vitro matured and unstimulated cycles oocytes by metaphase comparative genomic hybridization (mCGH)
- Author
-
Tahsin Yakut, Mutlu Karkucak, L. Keskintepe, Geoffrey Sher, Uludağ Üniversitesi/Tıp Fakültesi/Tıbbi Genetik Anabilim Dalı., Yakut, Tahsin, Karkucak, Mutlu, GIS-1493-2022, and FDJ-7858-2022
- Subjects
Oocyte ,Biochemistry & molecular biology ,Aneuploidy ,In vitro oocyte maturation techniques ,Embryo development ,Polar body ,Chromosome 11 ,Ploidy ,Maturation ,Cell differentiation ,Gonadotropin ,General Medicine ,Chromosome 15 ,Vivo ,Chromosome 13 ,Chorionic gonadotropin ,Chromosome 21 ,Chromosome 22 ,medicine.anatomical_structure ,Ivf ,Metaphase chromosome ,embryonic structures ,Female ,Chromosome aberration ,Embryo quality ,Human ,Adult ,In Vitro Studies ,Human Oocytes ,Intracytoplasmic Sperm Injections ,Polar body 1 ,Clinical article ,Cell cycle parameters ,In vivo oocyte maturation ,Biology ,Article ,Time ,Age ,In vivo ,Genetics ,medicine ,Humans ,Chromosome aberrations ,Molecular Biology ,Metaphase ,Comparative genomic hybridization ,urogenital system ,Embryos ,Methodology ,Follitropin ,medicine.disease ,Molecular biology ,Standardization ,In vitro maturation ,In vitro oocyte maturation ,Euploid ,Human cell ,Fertilization ,Stimulation ,Polar bodies ,Oocytes ,Comparative study ,Cell maturation ,Cytology ,Controlled study ,Unstimulated cell cycle - Abstract
A common observation after in vitro matured oocyte is that they yield poorer embryo quality compared to their in vivo counterparts. This study was designed to assess chromosomal status with metaphase comparative genomic hybridization after in vitro maturation (IVM) in unstimulated cycles and compare the results with those obtained after in vivo maturation. Patients without any obstetrical or gynecological pathology were admitted into the study. IVM oocytes were collected 36 h post hCG and matured in vitro at 37A degrees C in 5% O-2, 6% CO2, and 89% air for 36 h. All matured (metaphase II) oocytes were subject to polar body 1 (PB-1) biopsy and vitrified individually. PB-1 samples were transferred into 0.25 cc PCR tubes containing 2.5 mu l of PBS. PB-1 samples from 12 IVM patients were studied. Twenty-six out of 63 PB-1 samples (41%) were determined as euploid and 37 samples (59%) were aneuploid, whereas these values were 42% euploid and 58% aneuploid in the control group (in vivo matured oocytes). No statistical differences were found between the IVM and the control groups for euploid-aneuploid samples (P = 0.900). More aneuploidy was observed on chromosomes 11, 13, 15, 21, and 22 after IVM. Results show a non-significant rate of abnormal PB-1 formation after IVM compared to in vivo maturation. More aneuploidy was observed in chromosomes 11, 13, 15, 21, and 22 in the IVM group.
- Published
- 2012
39. Glucose-induced increase in circulating progenitor cells is blunted in polycystic amenorrhoeic subjects
- Author
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Soumi Bairagi, Subash Babu, Abel Arul Nathan, N. Pavan Kumar, Jayashree Gopal, and Madhulika Dixit
- Subjects
medicine.medical_treatment ,correlation analysis ,CD34 ,hyperandrogenism ,tertiary health care ,insulin resistance ,estrogen ,AC133 Antigen ,CD34 antigen ,endothelium cell ,CD133 antigen ,glucose ,Amenorrhea ,cell count ,Glucose tolerance test ,medicine.diagnostic_test ,adult ,Stem Cells ,Rehabilitation ,Obstetrics and Gynecology ,clinical controlled study ,Fasting ,Flow Cytometry ,Polycystic ovary ,medicine.anatomical_structure ,female ,luteinizing hormone ,follitropin ,diet restriction ,Polycystic Ovary Syndrome ,medicine.medical_specialty ,bone marrow ,Endothelium ,cell kinetics ,India ,glucose tolerance test ,Biology ,Insulin resistance ,fibronectin ,nitric oxide ,blood ,Antigens, CD ,Internal medicine ,ovary polycystic disease ,medicine ,glucose homeostasis ,Humans ,human ,Progenitor cell ,immunofluorescence ,cell marker ,protein expression ,Glycoproteins ,endothelial nitric oxide synthase ,cell culture ,Insulin ,human cell ,Estrogens ,case control study ,medicine.disease ,Frizzled Receptors ,stem cell ,cell differentiation ,Endocrinology ,Reproductive Medicine ,Case-Control Studies ,Peptides ,C peptide ,Homeostasis - Abstract
BACKGROUND Glucose-induced kinetics of bone marrow-derived stem cells in healthy females is presently unknown. The objectives of this study were to determine whether circulating levels of CD133(+), CD34(+) and CD133(+)CD34(+) cells increase in response to glucose load in healthy females and whether the kinetics is altered in amenorrhoeic women. The other objective of the work was to compare the endothelial differentiation potential of peripheral blood-derived endothelial progenitor cells (EPCs) from healthy versus amenorrhoeic women. METHODS In this case-control study, 44 amenorrhoeic subjects and 36 age-matched females with no menstrual disturbance were recruited at Apollo Hospitals, a Tertiary health care center in Chennai, India. Circulating bone marrow-derived stem cells were measured by two color direct flow cytometry. Cultured progenitor cells were characterized at Day 7 and 14 for expression of endothelial markers and production of nitric oxide (NO) via immunofluoroscence. RESULTS The amenorrhoeic subjects were insulin resistant with homeostatic model of assessment of insulin resistance values of 3.33 ± 0.3 versus 1.75 ± 0.148 observed for controls (P< 0.0001). Among the amenorrhoeic subjects, 38 subjects had polycystic ovaries with no signs of hyperandrogenism. Fasting levels of CD133(+), CD34(+) and CD133(+)CD34(+) cells were reduced in amenorrhoeic subjects (P< 0.001). There was a 1.5 to 2-fold increase in the circulating levels of these cells in response to 75 g oral glucose challenge at 1 and 2 h post-load conditions in controls, which was significantly blunted for CD133(+) (P< 0.001) and CD133(+)CD34(+) (P< 0.001) cells in amenorrhoeic subjects. A positive correlation was observed between estrogen and fasting CD133(+) (r= 0.205, P= 0.070), CD34(+) (r= 0.249, P= 0.027) and CD133(+)CD34(+) (r= 0.217, P= 0.055) cell counts. Additionally, fasting counts for CD34(+) and CD133(+)CD34(+) cells positively correlated with FSH and inversely correlated with LH and C-peptide in the polycystic group. Cultured cells from polycystic subjects exhibited reduced adherence to fibronectin and expressed lower levels of endothelial nitric-oxide synthase and NO. CONCLUSIONS Oral glucose-induced increase in circulating numbers of CD133(+) and CD133(+)CD34(+) cells and endothelial differentiation potential of peripheral blood-derived EPCs is attenuated in insulin resistant amenorrhoeic subjects.
- Published
- 2012
40. Receptor binding and functional properties of chimeric human follitropin prepared by an exchange between a small hydrophilic intercysteine loop of human follitropin and human lutropin
- Author
-
Xunxian Liu, James A. Dias, and Yiqiu Zhang
- Subjects
endocrine system ,medicine.medical_specialty ,Protein Conformation ,medicine.drug_class ,Recombinant Fusion Proteins ,Molecular Sequence Data ,Enzyme-Linked Immunosorbent Assay ,Follitropin ,Biology ,Binding, Competitive ,Biochemistry ,Protein Structure, Secondary ,Cell Line ,Radioligand Assay ,Structure-Activity Relationship ,Internal medicine ,medicine ,Humans ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,Progesterone ,Binding selectivity ,chemistry.chemical_classification ,luteinizing hormone/choriogonadotropin receptor ,Cell Biology ,Luteinizing Hormone ,Receptors, LH ,Peptide Fragments ,Amino acid ,Endocrinology ,chemistry ,Mutagenesis, Site-Directed ,Receptors, FSH ,Follicle Stimulating Hormone ,Gonadotropin ,Follicle-stimulating hormone receptor ,Glycoprotein ,Luteinizing hormone ,Baculoviridae ,hormones, hormone substitutes, and hormone antagonists ,Protein Binding - Abstract
The family of pituitary/placental glycoprotein hormones includes follicle-stimulating hormone (follitropin, FSH), luteinizing hormone (lutropin, LH), chorionic gonadotropin (choriogonadotropin, CG), and thyroid-stimulating hormone (thyrotropin, TSH). These glycoproteins are heterodimeric, with an alpha-subunit of identical primary structure and a hormone-specific beta-subunit which is believed to confer receptor specificity. Previous studies demonstrated that substitution of hFSH beta residues 88-108 in place of the carboxyl terminus of hCG beta, residues 94-145, conferred to human (h) CG an FSH receptor binding specificity. To more finely map the LH/FSH receptor binding specificity determinant, hFSH beta residues 88DSDS91, within the small hydrophilic intercysteine loop, and residues 95TVRGLG100 COOH-terminal to the loop were switched to hLH beta residues 94RRST97 and residues 101GGPKDH106, respectively. Substitution of hLH beta residues 94RRST97 in place of hFSH beta residues 88DSDS91 did not affect FSH receptor binding or activation but, remarkably, conferred LH receptor binding activity to the chimera. This chimera retained the ability to stimulate progesterone production in a Yl cell line which expresses human FSH receptor. Replacing hFSH beta residues 95TVRGLG100 with hLH beta residues 101GGPKDH106 diminished FSH receptor binding activity but did not confer LH receptor binding to the chimera. This study suggests that amino acids within hFSH beta residues 95TVRGLG100 are important for FSH binding specificity and that hLH beta residues 94RRST97 are involved in LH receptor binding specificity. Thus, although the hFSH beta and hLH beta subunits may fold similarly, the loci of receptor binding specificity are not entirely homologous.
- Published
- 1994
41. Influence of temperature regime on endocrine parameters and vitellogenesis during experimental maturation of European eel (Anguilla anguilla) females
- Author
-
Sylvie Baloche, Juan F. Asturiano, G. van den Thillart, Luz Pérez, Arjan P. Palstra, Sylvie Dufour, and David S. Peñaranda
- Subjects
Pituitary gland ,Follitropin beta subunit ,Ovary development ,PRODUCCION ANIMAL ,Gene ,Animal tissue ,Vitellogenins ,Endocrinology ,E2 ,Vitellogenin blood level ,Maturation ,MRNA ,Protein blood level ,Japanese eel ,Vitellogenin 2 gene ,Fshß ,Priority journal ,0303 health sciences ,Luteinizing hormone beta subunit ,biology ,Estradiol ,Anguilla anguilla ,induced sexual-maturation ,Esr1 ,Vitellogenesis ,Eel ,Temperature ,04 agricultural and veterinary sciences ,Wageningen Marine Research ,Estradiol blood level ,Lh beta gene ,medicine.anatomical_structure ,BIOLOGIA ANIMAL ,Liver ,Pituitary Gland ,Follicle Stimulating Hormone, beta Subunit ,Female ,Estrogen receptor 1 gene ,Luteinizing hormone ,Lhß ,glycoprotein hormone alpha ,Ovulation ,medicine.medical_specialty ,Carps ,Endocrine System ,luteinizing-hormone ,Vitellogenin ,Heat treatment ,Article ,03 medical and health sciences ,Internal medicine ,medicine ,artificial maturation ,Endocrine system ,Animals ,14. Life underwater ,Animal experiment ,Carp ,Fsh beta gene ,ii-beta subunits ,030304 developmental biology ,conger conger-myriaster ,messenger-rna levels ,goldfish carassius-auratus ,Tissue Extracts ,Developmental stage ,Vtg2 ,Follitropin ,biology.organism_classification ,Nonhuman ,Anguilla ,Hormone ,japanese eel ,040102 fisheries ,biology.protein ,0401 agriculture, forestry, and fisheries ,Animal Science and Zoology ,Hypophysis extract ,Controlled study ,Nucleotide sequence ,trout oncorhynchus-mykiss - Abstract
[EN] We examined the effect of temperature in European silver eels during their maturation induced by injections of carp pituitary extract on endocrine parameters: pituitary fshß and lhß expression, plasma 17ß-estradiol (E2) and vitellogenin, estrogen receptor 1 (esr1), and vitellogenin 2 (vtg2) expression in liver. A variable thermal regime (T10) that increased from 10° to 14° and 17 °C was compared with a constant 20 °C regime (T20) during 12. weeks. T10 caused a faster development until week 8, higher fshß, lhß, esr1 expression, and higher E2 levels. The results strongly suggest that T10 is inducing a higher endogenous FSH level which increases the E2 circulating level during vitellogenesis. A variable thermal regime induced an fshß expression and E2 profile in vitellogenic hormonally matured eel females that were more similar to the profile observed in other naturally maturing fish. © 2011 Elsevier Inc., This study was funded from the European Community’s 7th Framework Programme under the Theme 2 ‘‘Food, Agriculture and Fisheries, and Biotechnology’’, Grant Agreement No. 245257 (Pro-Eel) and Generalitat Valenciana through the GV/2007/202 project and ACOMP/2011/229. D.S. Peñaranda has a postdoc grant from the Universitat Politècnica de València (CE-01-10). The authors would like to thank M. Nieveen for technical assistance.
- Published
- 2011
42. Development of third-generation immunochemiluminometric assays of follitropin and lutropin and clinical application in determining pediatric reference ranges
- Author
-
E Carlton, W D Odell, Murugan R. Pandian, and D A Fisher
- Subjects
endocrine system ,medicine.medical_specialty ,Immunoradiometric assay ,medicine.drug_class ,Biochemistry (medical) ,Clinical Biochemistry ,Radioimmunoassay ,Follitropin ,Biology ,medicine.disease ,Lower limit ,Third generation ,Biological fluid ,Endocrinology ,Internal medicine ,medicine ,Precocious puberty ,Gonadotropin ,hormones, hormone substitutes, and hormone antagonists - Abstract
We developed dioxatane-based immunochemiluminometric assays (ICMAs) for lutropin (LH) and follitropin (FSH), using monoclonal antibodies. These ICMAs have a minimal detectable dose (analytical sensitivity) of 0.01 IU/L, extending the lower limit of sensitivity 10-fold (from 0.10 IU/L) when compared with immunoradiometric assays (IRMA) (second generation), and thus provide a true third-generation assay. Daytime FSH and LH concentrations were measured in 236 boys and 195 girls. Unlike the previous assays, all the samples had detectable concentrations of LH and FSH. In agreement with results from earlier methods, the present results indicate that for both sexes mean FSH and LH concentrations are relatively high during the early months of life, fall to baseline prepubertal concentrations by 12-18 months, and remain low until the onset of puberty. During puberty, the mean concentrations of FSH and LH increase significantly in both girls and boys with each stage of puberty, but there is considerable overlap between stages. These third-generation FSH and LH ICMAs reliably separate daytime plasma FSH and LH concentrations of prepubertal children from those of sexually mature children, and therefore can more reliably distinguish between the major causes of precocious puberty (e.g., gonadotropin dependent and independent). Our LH assay is also useful in monitoring the gonadotropin-releasing hormone therapy of patients with gonadotropin-dependent precocious puberty.
- Published
- 1993
43. The flanking amino acids of the human follitropin beta-subunit 33-53 region are involved in assembly of the follitropin heterodimer
- Author
-
Cheng Liu, K. E. Roth, J. A. Dias, J. B. Shaffer, and B. A. L. Shepard
- Subjects
Protein Folding ,medicine.medical_specialty ,Immunoprecipitation ,Protein subunit ,Molecular Sequence Data ,Mutant ,Follitropin ,Biology ,Cell Line ,Endocrinology ,Internal medicine ,Complementary DNA ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Polyacrylamide gel electrophoresis ,Peptide sequence ,chemistry.chemical_classification ,Base Sequence ,Molecular biology ,Recombinant Proteins ,Amino acid ,chemistry ,Mutation ,Mutagenesis, Site-Directed ,Follicle Stimulating Hormone ,Oligonucleotide Probes - Abstract
Previous analyses of the topology of human follitropin (hFSH) with monoclonal antibodies and antipeptide antibodies have led to a current operating hypothesis that some amino acids within the hFSH beta 33-53 region are surface oriented, and others participate in subunit contact. Protein structural analysis predicts beta-turns within this region, and the immunochemical studies indicate that the ends may be involved in subunit contact. In this study, hFSH beta was mutagenized to change 34TRDL37 to 34AAAA37 or 48QKTCT52 to 48AAACA52, allowing us to study the ends of the hFSH beta 33-53 sequence contiguous with the hFSH beta sequence. Wild-type and mutant cDNAs were coexpressed with alpha-subunit cDNA in CHOPro-5 cells. Wild-type hFSH was secreted from cells cotransfected with wild-type hFSH alpha and hFSH beta cDNAs, as expected. However, heterodimeric hFSH was minimally detected in the medium from cells transfected with the 34TRDL37 mutant and was not detected in the case of the 48QKTCT52 mutant. Analysis of cell lysates (intracellular FSH) by immunoprecipitation and polyacrylamide gel electrophoresis showed that wild-type and mutant beta-subunits were indistinguishable and recoverable intact from each cell line. Additionally, analysis of lysates with a conformation-specific monoclonal antibody 3G3 revealed that similar levels of properly folded beta-subunit were produced in cells expressing wild-type or either mutated beta-subunit. These data indicate that the flanking amino acids of the hFSH beta 33-53 region, in particular 48QKTCT52, are critical for assembly of hFSH heterodimer.
- Published
- 1993
44. Specificity and Detection Limit of Ten Pregnancy Tests
- Author
-
Ulf-Håkan Stenman, Henrik Alfthan, Aila Tiitinen, and Ulla-Maj Björses
- Subjects
Pregnancy test ,Detection limit ,endocrine system ,medicine.medical_specialty ,Pregnancy ,biology ,urogenital system ,Clinical Biochemistry ,Follitropin ,Urine ,General Medicine ,medicine.disease ,Human chorionic gonadotropin ,Endocrinology ,Internal medicine ,medicine ,biology.protein ,Beta (finance) ,reproductive and urinary physiology ,hormones, hormone substitutes, and hormone antagonists ,ATP synthase alpha/beta subunits - Abstract
Pregnancy tests have sufficiently low detection limits (25-200 IU/L of human chorionic gonadotropin, hCG) and good specificity as judged by lack of cross-reaction with lutropin (LH) and follitropin (FSH). However, little information is available on the specificity of the tests for subunits and fragments of hCG. The dominating form of hCG immunoreactivity in pregnancy urine is actually a 10 kD fragment of the beta chain of hCG called the core fragment. We have evaluated ten commonly used pregnancy test with respect to specificity for various forms of hCG and detection limit. Our results show that the detection limits of the tests are close to that claimed by the manufacturers and that the main form of hCG immunoreactivity detected is intact hCG. Four of the tests also measure the free beta subunit of hCG and one method also the hCG beta core fragment, but the detection limits were 7-70 fold those for hCG.
- Published
- 1993
45. Sexual dysfunction in obese and overweight women
- Author
-
S Tekekoglu, Fulya Akin, and Guzin Fidan Yaylali
- Subjects
Questionnaires ,obesity ,Female sexual dysfunction ,Personal Satisfaction ,Overweight ,Body Mass Index ,thyrotropin ,thyroxine blood level ,Sex hormone-binding globulin ,Waist–hip ratio ,follitropin blood level ,Sex Hormone-Binding Globulin ,Surveys and Questionnaires ,estrogen ,orgasm ,Insulin ,Testosterone ,Sexual Dysfunctions, Psychological ,pathophysiology ,prasterone sulfate ,Adiposity ,clinical article ,anthropometry ,biology ,adult ,article ,satisfaction ,sex hormone binding globulin ,testosterone blood level ,medical history ,body fat ,prolactin blood level ,female ,waist hip ratio ,priority journal ,sexual dysfunction ,luteinizing hormone ,Female ,medicine.symptom ,follitropin ,Adult ,prolactin ,medicine.medical_specialty ,Urology ,hormone ,complication ,blood ,estradiol ,Internal medicine ,medicine ,Humans ,controlled study ,hydrocortisone ,human ,Obesity ,Orgasm ,thyroxine ,Estrogens/blood ,Hormones/blood ,Insulin/blood ,Obesity/*complications/physiopathology ,Overweight/*complications/physiopathology ,Sex Hormone-Binding Globulin/analysis ,Sexual Dysfunctions, Psychological/*epidemiology/etiology ,Testosterone/blood ,Waist-Hip Ratio ,business.industry ,questionnaire ,disease association ,Testosterone (patch) ,Estrogens ,case control study ,medicine.disease ,body mass ,Hormones ,thyrotropin blood level ,Endocrinology ,Sexual dysfunction ,female sexual dysfunction ,estradiol blood level ,biology.protein ,luteinizing hormone blood level ,business ,Sexual function ,Body mass index - Abstract
Both overweight and obesity have been identified as risk factors for sexual dysfunction in men, but the relationship between sexual function and amount of body fat in females is still obscure. There are few reported studies in women assessing the relationship between female sexual function index (FSFI) and body weight. The aim of this study was to identify the frequency of female sexual dysfunction (FSD) among obese and overweight women. A total of 45 obese and overweight and 30 age-matched voluntary healthy women serving as a control group were evaluated by a detailed medical and sexual history, including the FSFI questionnaire. Serum prolactin, cortisol, luteinizing hormone (LH), follicle-stimulating hormone (FSH), dehydroepiandrosterone-SO(4) (DHEA-S), testosterone, estradiol and sex hormone-binding globulin (SHBG) levels were measured. No significant difference was observed between controls and patients in terms of the FSH, LH, estradiol, free thyroxine and thyrotropin (TSH), testosterone and DHEA-S levels. The comparison of total FSFI scores between patients and controls showed no significant difference (P = 0.74). As the FSFI score of
- Published
- 2010
46. Design of a long-acting follitropin agonist by fusing the C-terminal sequence of the chorionic gonadotropin beta subunit to the follitropin beta subunit
- Author
-
P. S. Lapolt, Irving Boime, K. Nishimori, Aaron J. W. Hsueh, Fuad Fares, and N. Suganuma
- Subjects
Male ,endocrine system ,medicine.medical_specialty ,Metabolic Clearance Rate ,medicine.drug_class ,Recombinant Fusion Proteins ,Protein subunit ,Restriction Mapping ,Peptide ,Follitropin ,CHO Cells ,Protein Engineering ,Transfection ,Chorionic Gonadotropin ,Aromatase ,Cricetinae ,Internal medicine ,Testis ,medicine ,Animals ,Chorionic Gonadotropin, beta Subunit, Human ,Peptide sequence ,Cells, Cultured ,G alpha subunit ,chemistry.chemical_classification ,Granulosa Cells ,Multidisciplinary ,biology ,Chimera ,Organ Size ,Peptide Fragments ,Rats ,Endocrinology ,chemistry ,Drug Design ,Chorionic gonadotropin beta ,biology.protein ,Receptors, FSH ,Biological Assay ,Female ,Follicle Stimulating Hormone ,Gonadotropin ,hormones, hormone substitutes, and hormone antagonists ,ATP synthase alpha/beta subunits ,Half-Life ,Research Article - Abstract
Follitropin (FSH) is a pituitary glycoprotein hormone that is essential for the development of ovarian follicles and testicular seminiferous tubules. FSH is used clinically to stimulate follicular maturation for in vitro fertilization and treatment of anovulatory women. One issue regarding the clinical use of FSH is its short half-life in the circulation. To address this point, we constructed chimeric genes containing the sequence encoding the C-terminal peptide of the chorionic gonadotropin beta subunit (CG beta) fused to the translated sequence of the human FSH beta subunit (FSH beta). This region of CG beta is important for maintaining the prolonged plasma half-life of human CG dimer. The presence of the C-terminal peptide sequence did not significantly affect assembly of FSH beta with the alpha subunit or secretion of the dimer. In vitro receptor binding and steroidogenic activity of dimer bearing the FSH beta-C-terminal peptide chimera were the same as wild-type FSH. However, both the in vivo potency and half-life in circulation of the dimer bearing either one or two C-terminal peptide units were enhanced. Dimers containing FSH beta-CG beta chimeras could serve as potent FSH agonists for clinical use, and the present strategy may have wide applications for enhancing the in vivo half-life of diverse proteins.
- Published
- 1992
47. Amphibian lutropin and follitropin from the bullfrog Rana catesbeiana. Complete amino acid sequence of the alpha subunit
- Author
-
Hiroaki Hayashi, Tomoko Hayashi, and Yoichi Hanaoka
- Subjects
Ovulation ,endocrine system ,Arginine ,Xenopus ,Molecular Sequence Data ,Follitropin ,Biology ,Biochemistry ,Bullfrog ,Sequence Homology, Nucleic Acid ,Animals ,Trypsin ,Amino Acid Sequence ,Cyanogen Bromide ,Peptide sequence ,Chromatography, High Pressure Liquid ,G alpha subunit ,chemistry.chemical_classification ,Rana catesbeiana ,Base Sequence ,Ovary ,Nucleic acid sequence ,Luteinizing Hormone ,Amino acid ,chemistry ,Female ,Follicle Stimulating Hormone ,hormones, hormone substitutes, and hormone antagonists ,Cysteine - Abstract
The amino acid sequence of the alpha-subunit of the gonadotropins, lutropin and follitropin from bullfrog, Rana catesbeiana, has been determined. The alpha-subunit was identified in both hormones by the amino acid composition and ovulation activity of lutropin in the Xenopus ovary, by means of reconstituted hormones in various combinations. The amino acid sequences of two identical alpha-subunits from lutropin and follitropin were determined or deduced by different strategies. The alpha-subunit of those gonadotropins have 97 amino acid residues, the longest among the known alpha-subunits of gonadotropins, and one arginine insertion at position 29. Ten cysteine residues and two sugar-chain binding sites at Asn57 and Asn83 are completely conserved among the species. The molecular mass of this subunit is 11,026 Da not including the sugar chains. The bullfrog alpha-subunit has approximately 70% sequence identity with mammalian alpha-subunits.
- Published
- 1992
48. Differential appearance of the subunits of glycoprotein hormones (LH, FSH, and TSH) in the pituitary of bullfrog (Rana catesbeiana) larvae during metamorphosis
- Author
-
Makoto Sakai, Min Kyun Park, Shigeyasu Tanaka, and Kazumasa Kurosumi
- Subjects
endocrine system ,medicine.medical_specialty ,Pituitary gland ,media_common.quotation_subject ,Radioimmunoassay ,Thyrotropin ,Follitropin ,Gonadotropic cell ,Immunoenzyme Techniques ,Endocrinology ,Bullfrog ,Thyrotropic cell ,Internal medicine ,medicine ,Animals ,Metamorphosis ,media_common ,Rana catesbeiana ,biology ,Metamorphosis, Biological ,Luteinizing Hormone ,medicine.anatomical_structure ,Polyclonal antibodies ,Larva ,Pituitary Gland ,biology.protein ,Animal Science and Zoology ,Follicle Stimulating Hormone ,hormones, hormone substitutes, and hormone antagonists ,Hormone - Abstract
The ontogeny of gonadotrophs and thyrotrophs in the bullfrog pituitary was examined immunohistochemically using monoclonal antibodies against bullfrog lutropin beta-subunit (LH beta), follitropin beta-subunit (FSH beta) and its alpha-subunit, and polyclonal anti-human thyrotropin beta-subunit (TSH beta) serum. Immunoreactive alpha-subunit- and TSH beta-, but not FSH beta- and LH beta-containing cells were observed at embryonic stage 24 (Shumway's classification). Immunoreactive FSH beta cells first appeared at Taylor-Korllos stage V, and immunoreactive LH beta cells at stage X. Throughout metamorphosis, several gonadotrophs containing both FSH and LH were found in the ventrocaudal region, but most gonadotrophs contained only FSH. Immunoreactive alpha-subunit cells were always more frequent than the sum of immunoreactive beta-subunit cells, which was confirmed by quantitative studies using immunohistochemical and RIA techniques.
- Published
- 1991
49. Expression of biologically active heterodimeric bovine follicle-stimulating hormone in milk of transgenic mice
- Author
-
Jeffrey M. Rosen, Jerry J. Reeves, Joseph W. Anderson, Keiji Nishimori, Darrell N. Ward, David D. Deavila, Norman M. Greenberg, and Aaron J. W. Hsueh
- Subjects
Genetically modified mouse ,endocrine system ,Transgene ,Protein subunit ,Genetic Vectors ,Molecular Sequence Data ,Oligonucleotides ,Gene Expression ,Mice, Transgenic ,Follitropin ,Biology ,Polymerase Chain Reaction ,Mice ,Follicle-stimulating hormone ,Mammary Glands, Animal ,Gene expression ,Animals ,Lactation ,Secretion ,RNA, Messenger ,Cloning, Molecular ,Multidisciplinary ,Base Sequence ,Caseins ,DNA ,Blotting, Northern ,Molecular biology ,Blotting, Southern ,Milk ,Cattle ,Follicle Stimulating Hormone ,Research Article ,Hormone - Abstract
Follicle-stimulating hormone (FSH; follitropin) is a pituitary glycoprotein composed of two post-translationally modified subunits, which must properly assemble to be biologically active. FSH has been difficult to purify and to obtain in quantities sufficient for detailed biochemical studies. We have targeted FSH expression to the mammary gland of transgenic mice by using cDNAs encoding the bovine alpha and FSH beta subunits and a modified rat beta-casein gene-based expression system. Lines of bigenic mice expressing both subunits have been generated either by coinjection of the subunit transgenes or by mating mice that acquired and expressed transgenes encoding an individual subunit. Up to 60 international units (15 micrograms) of biologically active FSH per ml was detected in the milk of the bigenic mice. These lines provide a model system for studying the post-transcriptional mechanisms that effect the expression and secretion of this heterodimeric hormone.
- Published
- 1991
50. Development of a radioligand receptor assay for measuring follitropin in serum: application to premature ovarian failure
- Author
-
Ruth Freeman, Randall W. Whitcomb, Janet E. Hall, Patrick M. Sluss, Alan L. Schneyer, and William F. Crowley
- Subjects
endocrine system ,medicine.medical_specialty ,medicine.drug_class ,media_common.quotation_subject ,Biochemistry (medical) ,Clinical Biochemistry ,Follitropin ,Radioimmunoassay ,Luteal phase ,Biology ,Endocrinology ,Internal medicine ,Follicular phase ,Radioligand ,medicine ,Gonadotropin ,Follicle-stimulating hormone receptor ,hormones, hormone substitutes, and hormone antagonists ,Menstrual cycle ,media_common - Abstract
We have developed a radioligand receptor assay (RRA) with sufficient sensitivity and specificity for quantifying follitropin (FSH) in unextracted serum samples. Standard curves prepared by adding pituitary FSH to either buffer or gonadotropin-free serum were parallel and statistically indistinguishable in this assay, whereas gonadotropin-free serum alone had no activity. Cross-reactivity with related pituitary hormones was negligible. Pituitary FSH was calibrated with commonly used reference preparations so that RRA results could be compared with RIA results for identical standards. The patterns in daily blood samples in six normal menstrual cycles were similar by both methods. The mean RIA:RIA ratio in both the follicular and luteal phases was between 0.6 and 0.7, and at mid-cycle decreased to 0.48, suggesting an alteration of isohormone composition at mid-cycle. In 27 women with premature ovarian failure, RRA:RIA ratios ranged from below the RRA minimum detectable dose to 4.6, suggesting that immunoreactive FSH might not be capable of binding to the FSH receptor in some patients, whereas in patients with high RRA:RIA ratios, circulating inhibitors of FSH receptor binding might be present and perhaps contributing to the observed ovarian failure. Use of this RRA in conjunction with RIA and in vitro bioassays may better define the relative contribution of FSH isohormones, autocrine or paracrine modulators of FSH bioactivity, and FSH-receptor binding competitors to the "total FSH biological signal" as detected by the gonadal FSH receptor.
- Published
- 1991
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