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1.
Endocrinology ; 161(1)2020 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-31865385

RESUMO

Follicle-stimulating hormone (FSH) is a dimeric glycoprotein secreted by the anterior pituitary gonadotrope that is necessary for reproductive function in mammals. FSH primarily regulates granulosa cells and follicular growth in females, and Sertoli cell function in males. Since its identification in the 1930s and sequencing in the 1970s, significant progress has been made in elucidating its regulation and downstream function. Recent advances provide deeper insight into FSH synthesis, and effects in the gonads suggest potential roles in extragonadal tissues and examine pharmacological approaches and clinical applications in infertility treatment that now affect 18% of couples. These advances were discussed in detail in a number of reviews published in the last 2 years in Endocrinology. In this brief commentary, we summarize these reviews and point to the outstanding questions that should be answered in the near future to bridge a gap in our understanding of this hormone.


Assuntos
Hormônio Foliculoestimulante/genética , Regulação da Expressão Gênica , Células da Granulosa/metabolismo , Processamento de Proteína Pós-Traducional , Células de Sertoli/metabolismo , Animais , Feminino , Hormônio Foliculoestimulante/metabolismo , Humanos , Masculino , Folículo Ovariano/citologia , Folículo Ovariano/metabolismo , Ligação Proteica , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , Receptores do FSH/metabolismo , Literatura de Revisão como Assunto , Transdução de Sinais , Espermatogênese/genética
2.
Ann Endocrinol (Paris) ; 80(4): 234-239, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31439307

RESUMO

OBJECTIVE: Congenital FSH deficiency is an exceptional cause of male infertility most often attributed to FSH ß gene mutations. The few published cases report azoospermia, severe testicular hypotrophy and normal testosterone levels associated with normal virilization. We report the exploration of two young men aged 26 and 27 years with severe sperm abnormalities, moderate testicular hypotrophy and isolated FSH deficiency. METHODS: Several FSH, LH, total testosterone and inhibin B assays and FSH ß gene sequencing were performed. RESULTS: FSH was almost undetectable at baseline and poorly responsive to GnRH test, whereas LH was normal at baseline and increased after GnRH test. Testosterone levels were within the adult range, while inhibin B levels were upper-normal to high. No FSH ß gene mutations were found. Exogenous FSH treatment was followed by spontaneous pregnancy in one case and required intra-cytoplasmic sperm injection (ICSI) in the other. CONCLUSIONS: The paradoxical high levels of inhibin B reflect the presence of functional Sertoli cells and may explain the isolated FSH deficiency. An intra-gonadal factor stimulating inhibin B secretion is discussed.


Assuntos
Subunidade beta do Hormônio Folículoestimulante/genética , Hormônio Foliculoestimulante/deficiência , Infertilidade Masculina/diagnóstico , Oligospermia/diagnóstico , Adulto , Análise Mutacional de DNA , Hormônio Foliculoestimulante/genética , Humanos , Infertilidade Masculina/genética , Masculino , Mutação , Oligospermia/genética
3.
Mar Biotechnol (NY) ; 21(5): 697-706, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31372794

RESUMO

The availability of sexually mature fish often dictates the success of its captive breeding. In this study, we induced reproductive development in juvenile protogynous tiger grouper through oral administration of a plasmid (p) containing an engineered follicle-stimulating hormone (FSH). An expression construct (pcDNA3.1) was designed to express a single-chain FSH consisting of giant grouper FSH ß-subunit and glycoprotein subunit-α (CGα), linked by the carboxy-terminal peptide (CTP) sequence from the human chorionic gonadotropin (hCG). Single oral delivery of pFSH encapsulated in liposome and chitosan to tiger grouper yielded a significant increase in plasma FSH protein level after 4 days. Weekly pFSH feeding of juvenile tiger groupers for 8 weeks stimulated ovarian development as indicated by a significant increase in oocyte diameter and progression of oocytes to cortical alveolar stage. As the pFSH treatment progressed from 20 to 38 weeks, female to male sex change was initiated, characterized by oocyte regression, proliferation of spermatogonial cells, and occurrence of spermatogenic cysts. It was also associated with significantly lower mRNA expression of steroidogenic genes (cyp11b, cyp19a1a, and foxl2) and basal plasma levels of sex steroid hormones 17ß-estradiol (E2), testosterone (T), and 11-ketotestosterone (11KT). Results suggest that pFSH stimulates ovarian development up to cortical alveolar stage and then initiates sex change in tiger grouper. These findings significantly contribute to our knowledge on the role of FSH in the development of protogynous hermaphroditic fish. This study is the first to demonstrate induction of reproductive development in fish through oral delivery of plasmid gonadotropin.


Assuntos
Gonadotropina Coriônica/genética , Hormônio Foliculoestimulante/genética , Gônadas/efeitos dos fármacos , Organismos Hermafroditas/efeitos dos fármacos , Perciformes/genética , Processos de Determinação Sexual/efeitos dos fármacos , Diferenciação Sexual/efeitos dos fármacos , Administração Oral , Animais , Quitosana/química , Gonadotropina Coriônica/administração & dosagem , Gonadotropina Coriônica/biossíntese , Composição de Medicamentos , Feminino , Proteínas de Peixes/biossíntese , Proteínas de Peixes/genética , Hormônio Foliculoestimulante/administração & dosagem , Hormônio Foliculoestimulante/biossíntese , Hormônios Esteroides Gonadais/biossíntese , Hormônios Esteroides Gonadais/genética , Gônadas/crescimento & desenvolvimento , Gônadas/metabolismo , Organismos Hermafroditas/genética , Humanos , Lipossomos/administração & dosagem , Lipossomos/química , Masculino , Oogênese/efeitos dos fármacos , Oogênese/genética , Perciformes/crescimento & desenvolvimento , Perciformes/metabolismo , Plasmídeos/química , Plasmídeos/metabolismo , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Pré-Seleção do Sexo/métodos , Espermatogênese/efeitos dos fármacos , Espermatogênese/genética
4.
BMC Res Notes ; 12(1): 455, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31340838

RESUMO

OBJECTIVE: Study analyzes mutation in mtDNA (Mitochondrial DNA) among diabetic women with PCOS in non-diabetic diabetic women and compared with the healthy control. Women with known case of hyperandrogenism, ovulatory dysfunction and/or polycystic ovaries were selected and anthropometric and demographic variables were collected during their clinical visit. Biochemical estimation of glucose, FSH, LH, estradiol (E2), and insulin levels were analyzed. Mutational analysis of mt-tRNA genes of each individual was compared with the updated consensus Cambridge sequence. The mtDNA content was determined in triplicate using SYBR green PCR mastermix. RESULTS: The clinical and biochemical characteristics of participants showed no statistical difference in age and/or FSH, PRL, E2, PRGE or fasting glucose value between patients of different groups. Women with PCOS-D had significantly higher LH, LH/FSH, TT and fasting insulin levels and HOMA-IR with respect to the control group. Ten different type of mutation were seen in POCS group. Most of these mutations were confined to evolutionarily conserved region. The mtDNA copy numbers were considerably lower PCOS group irrespective of diabetic status. To conclude, the current study inferred that the mutations occur in the mitochondrial genome, mt-tRNA in specific, are the important causal factor in PCOS.


Assuntos
DNA Mitocondrial/genética , Diabetes Mellitus Tipo 2/genética , Hiperandrogenismo/genética , Mitocôndrias/genética , Síndrome do Ovário Policístico/genética , RNA de Transferência/genética , Adulto , Glicemia/metabolismo , Estudos de Casos e Controles , Variações do Número de Cópias de DNA , DNA Mitocondrial/sangue , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/patologia , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Foliculoestimulante/genética , Expressão Gênica , Humanos , Hiperandrogenismo/sangue , Hiperandrogenismo/complicações , Hiperandrogenismo/patologia , Insulina/sangue , Resistência à Insulina , Hormônio Luteinizante/sangue , Hormônio Luteinizante/genética , Herança Materna , Mitocôndrias/patologia , Ovário/metabolismo , Ovário/patologia , Síndrome do Ovário Policístico/sangue , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/patologia , Estudos Prospectivos , RNA de Transferência/sangue
5.
J Assist Reprod Genet ; 36(6): 1185-1194, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31079267

RESUMO

PURPOSE: Follicle-stimulating hormone receptor (FSHR) expression in granulosa cells is critical in enabling follicles to achieve accelerated growth. Although FSHR expression has been reported to be epigenetically regulated, the mechanism is unclear. Cooperation between oocytes and granulosa cells is also essential for normal follicular growth. Among oocyte-derived factors, bone morphogenetic protein 15 (BMP15) promotes follicular growth and is suggested to have epigenetic effects. We examined the role of BMP15 in the acquirement of FSHR in human granulosa cells. METHODS: Immortalized non-luteinized human granulosa (HGrC1) cells were stimulated with trichostatin A (TSA) or BMP15 to analyze FSHR expression, histone modifications, and USF1/2 binding at the FSHR promoter region. Histone acetyl transferase (HAT) activity and phosphorylation of Smad 1/5/8 and p38 MAPK were examined with or without BMP15, SB203580, and LDN193189. CYP19A1 expression and estradiol production were also studied. RESULTS: TSA and BMP15 induced FSHR mRNA expression in a dose-dependent manner and histone modifications were observed with increased binding of USF1/2. BMP15 increased FSHR protein expression, which was suppressed by LDN193189. BMP15 increased phosphorylation of Smad 1/5/8 and significantly increased HAT activity, which was inhibited by LDN193189, but not by SB203580. BMP15 increased phosphorylation of p38 MAPK and USF1. LDN193189 suppressed BMP15-induced phosphorylation of both p38 MAPK and USF1, whereas SB203580 suppressed the phosphorylation of USF1. BMP15 increased CYP19A1 mRNA expression and estradiol production. CONCLUSION: BMP15 induced FSHR expression in human granulosa cells through Smad and non-Smad pathways. This mechanism of FSHR induction by BMP15 may be utilized for controlling follicular growth.


Assuntos
Proteína Morfogenética Óssea 15/farmacologia , Células da Granulosa/metabolismo , Oócitos/crescimento & desenvolvimento , Receptores do FSH/genética , Aromatase/genética , Estradiol/genética , Feminino , Hormônio Foliculoestimulante/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Células da Granulosa/patologia , Humanos , Ácidos Hidroxâmicos/farmacologia , Oócitos/metabolismo , Folículo Ovariano/crescimento & desenvolvimento , Folículo Ovariano/metabolismo , Pirazóis/farmacologia , Pirimidinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Proteínas Smad/genética
6.
J Biol Regul Homeost Agents ; 33(2): 461-468, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30968676

RESUMO

Endocrinal interactions are one of the most crucial regulatory mechanisms that maintain the state of homeostasis in humans. Processes such as oogenesis, folliculogenesis, menstruation and pregnancy remain under hormonal control. A key role in folliculogenesis is played by granulosa cells. Moreover, granulosa cells take part in corpus luteum formation after ovulation. Because of that, it is important to understand the ways in which the granulosa cells, associated with those processes, respond to hormonal stimulus. In the present study, a transcriptomic analysis of human granulosa cells (GCs) was carried out with the use of expression microarrays. The results were validated by RT-qPCR. The total RNA was isolated after 1st, 7th, 15th and 30th days of long-term primary cultures. The main focus of this work was placed on the genes belonging to "Response to estradiol", "Response to follicle-stimulating-hormone", "Cellular response to hormone stimulus", "Cellular hormone metabolic process" and "Hormone biosynthetic process" gene ontology groups. These groups of genes have been associated with GC hormone metabolism and cellular response to hormones. Eighty genes belonging to these groups were identified. Those that were members of more than one of the analyzed gene ontology groups, or exhibited unique expression patterns, were selected for further analysis. All of the selected genes were described, with their expression patterns detailed. In this manuscript, two gene expression patterns have been described. The first one showed large downregulation of genes in the later stages of culture, with the second one presenting upregulation of expression after day 1 of IVC. The present research was focused on six genes found to be the most important for steroidogenesis: STAR, POR, CYP11A1, ADM, GCLC, IL1B, as well as three genes of higher expression at the later stages of long-term in vitro culture: NR2F2, BMP4, COL1A1. The main goal of the presented study was to select genes involved in response to hormonal stimulus and hormone metabolism in GC long-term in vitro culture.


Assuntos
Estradiol/genética , Hormônio Foliculoestimulante/genética , Células da Granulosa/metabolismo , Folículo Ovariano/crescimento & desenvolvimento , Células Cultivadas , Feminino , Humanos , Oogênese , Ovulação , Gravidez
7.
Anim Sci J ; 90(4): 473-480, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30793438

RESUMO

This study was aimed to address melatonin receptor expression, mRNA level of hypothalamus and hypophysis hormone receptors (GnRHR, FSHR, and LHR), steroidogenesis, cell cycle, apoptosis, and their regulatory factors after addition of melatonin for 24 hr in cultured buffalo granulosa cells (GCs). The results revealed that direct addition of different concentrations of melatonin (100 pM, 1 nM, and 100 nM) resulted in significant upregulation (p < 0.05) of mRNA level of melatonin receptor 1a (MT1) without affecting melatonin receptor 1b (MT2). Melatonin treatment significantly downregulated (p < 0.05) mRNA level of FSH and GnRH receptors, whereas 100 nM dose of melatonin significantly increased mRNA level of LH receptor. Treatment with 100 nM of melatonin significantly decreased the basal progesterone production with significant decrease (p < 0.05) in mRNA levels of StAR and p450ssc, and lower mRNA level of genes (Insig1, Lipe, and Scrab1) that affect cholesterol availability. Melatonin supplementation suppressed apoptosis (100 nM, p < 0.05) and enhanced G2/M phase (1 nM, 100 nM, p < 0.05) of cell cycle progression which was further corroborated by decrease in protein expression of caspase-3, p21, and p27 and increase in bcl2. Our results demonstrate that melatonin regulates gonadotrophin receptors and ovarian steroidogenesis through MT1. Furthermore, the notion of its incorporation in apoptosis and proliferation of buffalo GCs extends its role in buffalo ovaries.


Assuntos
Apoptose/efeitos dos fármacos , Estradiol/metabolismo , Hormônio Foliculoestimulante/metabolismo , Células da Granulosa/metabolismo , Células da Granulosa/patologia , Melatonina/farmacologia , Progesterona/metabolismo , Animais , Búfalos , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Hormônio Foliculoestimulante/genética , Expressão Gênica/efeitos dos fármacos , Melatonina/fisiologia , RNA Mensageiro/metabolismo , Receptor MT1 de Melatonina/genética , Receptor MT1 de Melatonina/metabolismo , Receptores do LH/genética , Receptores do LH/metabolismo , Receptores LHRH/metabolismo , Regulação para Cima/efeitos dos fármacos
8.
J Assist Reprod Genet ; 36(3): 425-432, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30610662

RESUMO

Infertility due to Gonadotropin-Resistant Ovary Syndrome (GROS) is a rare type of hypergonadotropic hypogonadism. Here, we report an original case of GROS, associated with compound heterozygous follicle-stimulating hormone receptor (FSHR) variants, in a woman who achieved a live birth by in vitro maturation (IVM) of her oocytes. This 31-year-old woman consulted our assisted reproduction center for a second opinion after having been advised, because of pervasive high serum follicle-stimulating hormone (FSH) levels, to pursue in vitro fertilization (IVF) with donor oocytes. She presented with primary infertility and progressively prolonged menstrual cycles. Her serum FSH levels were indeed found to be high, but in discordance with a normal anti-Müllerian hormone (AMH) level and antral follicle count. Genetic investigation found the patient to be compound heterozygous for two FSHR variants: I160T, a known pathologic variant, and N558H, which has never been previously reported. As there was no ovarian response to high daily doses of exogenous gonadotropins, IVM was proposed to the patient with success and she finally delivered at term a healthy boy. Effects of the receptor variants were analyzed in heterologous cells. Whereas the I160T mutation blocked FSHR membrane trafficking and FSH-stimulated cAMP-dependent signaling in transfected CHO cells, the novel variant, N558H, functioned equivalently to wild-type FSHR in the assays employed. In conclusion, IVM should always be offered as a first-line therapy to infertile women presenting with GROS. The N558H variant discovered in FSHR is novel, but its functional significance, if any, is unresolved and merits further investigation as it may be associated with a recessive FSHR-related disorder.


Assuntos
Hormônio Foliculoestimulante/genética , Técnicas de Maturação in Vitro de Oócitos , Insuficiência Ovariana Primária/genética , Receptores do FSH/genética , Adulto , Animais , Hormônio Antimülleriano/sangue , Células CHO , Cricetulus , Feminino , Hormônio Foliculoestimulante/sangue , Humanos , Nascimento Vivo , Oócitos/crescimento & desenvolvimento , Oócitos/patologia , Insuficiência Ovariana Primária/sangue , Insuficiência Ovariana Primária/patologia , Transdução de Sinais
9.
Genet Med ; 21(4): 930-938, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30224786

RESUMO

PURPOSE: Primary ovarian insufficiency (POI) is a frequent disorder that affects ~1% of women under 40 years of age. POI, which is characterized by the premature depletion of ovarian follicles and elevated plasma levels of follicle-stimulating hormone (FSH), leads to infertility. Although various etiological factors have been described, including chromosomal abnormalities and gene variants, most cases remain idiopathic. The aim of the present study was to identify and validate functionally new sequence variants in ATG (autophagy-related genes) leading to POI. METHODS: We have reanalyzed, in silico, the exome sequencing data from a previously reported work performed in 69 unrelated POI women. Functional experiments using a classical hallmark of autophagy, the microtubule-associated protein 1 light chain 3ß (LC3), were then used to link these genes to this lysosomal degradation pathway. RESULTS: We venture a functional link between ATG7 and ATG9A variants and POI. We demonstrated that variant ATG7 and ATG9A led to a decrease in autophagosome biosynthesis and consequently to an impairment of autophagy, a key biological process implicated in the preservation of the primordial follicles forming the ovarian reserve. CONCLUSION: Our results unveil that impaired autophagy is a novel pathophysiological mechanism involved in human POI.


Assuntos
Proteína 7 Relacionada à Autofagia/genética , Proteínas Relacionadas à Autofagia/genética , Autofagia/genética , Proteínas de Membrana/genética , Insuficiência Ovariana Primária/genética , Proteínas de Transporte Vesicular/genética , Adulto , Feminino , Hormônio Foliculoestimulante/genética , Predisposição Genética para Doença , Humanos , Mutação com Perda de Função/genética , Menopausa Precoce/genética , Insuficiência Ovariana Primária/patologia , Sequenciamento Completo do Exoma
10.
Biosci Rep ; 39(1)2019 01 31.
Artigo em Inglês | MEDLINE | ID: mdl-30463907

RESUMO

Large doses of flavonoids could cure many diseases with no serious side effects. However, the role of flavonoids in the treatment of polycystic ovary syndrome (PCOS) has not been reported. Therefore, total flavonoids extracted from Nervilia Fordii were selected to explore its therapeutic efficiency in PCOS. PCOS rat model was constructed to explore the role of total flavonoids in the treatment of PCOS. ELISA was used to assess the changes of ovulation function under the treatment of total flavonoids with or without exogenous interleukin-6 (IL-6). Western blot, real-time PCR and immunohistochemistry were carried out to assess the related molecular mechanisms. We explored that total flavonoids obviously increased the serum levels of follicle-stimulating hormone (FSH), and sharply decreased the serum levels of luteinizing hormone (LH), testosterone (T) and insulin (INS) in the PCOS-IR rats via partly inhibiting the activation of JAK2/STAT3 pathway, partially up-regulating the IL-6 expression and partially down-regulating the suppressor of cytokine signaling 3 (SOCS3) expression in ovaries of PCOS rats. The effect of total flavonoids on estrous cycles, serum levels of FSH, LH, T and INS were partially attenuated by IL-6 in PCOS rat model. Moreover, IL-6 significantly reversed the effect of total flavonoids on the phosphorylation of JAK2/STAT3, the expression of IL-6 and SOCS3 in ovaries of PCOS rats. Total flavonoids extracted from Nervilia Fordii might induce the expression of IL-6 in ovary and act as a potential therapeutic drug for the treatment of PCOS.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Flavonoides/farmacologia , Interleucina-6/genética , Janus Quinase 2/genética , Orchidaceae/química , Síndrome do Ovário Policístico/tratamento farmacológico , Fator de Transcrição STAT3/genética , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Glicemia , Gonadotropina Coriônica/administração & dosagem , Modelos Animais de Doenças , Ciclo Estral/efeitos dos fármacos , Feminino , Flavonoides/isolamento & purificação , Hormônio Foliculoestimulante/sangue , Hormônio Foliculoestimulante/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Insulina/administração & dosagem , Insulina/sangue , Resistência à Insulina , Interleucina-6/sangue , Interleucina-6/farmacologia , Janus Quinase 2/sangue , Hormônio Luteinizante/sangue , Hormônio Luteinizante/genética , Ovulação/efeitos dos fármacos , Extratos Vegetais/química , Síndrome do Ovário Policístico/induzido quimicamente , Síndrome do Ovário Policístico/genética , Síndrome do Ovário Policístico/patologia , Ratos , Ratos Sprague-Dawley , Fator de Transcrição STAT3/sangue , Transdução de Sinais , Testosterona/sangue , Testosterona/genética
11.
Genet Test Mol Biomarkers ; 23(1): 39-44, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30585745

RESUMO

BACKGROUND: Follicle-stimulating hormone (FSH) is essential to the hypothalamic-pituitary-gonadal axis, playing a key role in human reproduction. It is a heterodimer comprised of a hormone-specific ß-chain (FSH-ß) that is associated with an α-chain. It exerts its biological activities by binding to the FSH receptor (FSHR). The ß-subunit, which is encoded by the FSHB gene, is responsible for ensuring binding specificity to the FSHR. There is a promoter polymorphism in this gene, c.-211G>T (rs10835638), upstream of the transcription start site; and in vitro studies have reported that the T allele decreases FSHB transcription in gonadotrophic cells. AIMS: Investigate the possible effects of the FSHB c.-211G/T polymorphism on hormonal profile and in in vitro fertilization (IVF)/intracytoplasmic sperm injection outcomes in normoovulatory Brazilian women. METHODS: A cross-sectional study of 140 women (median age = 33 years [CI: 32-34]) with infertility mainly caused by male (n = 85) or tuboperitoneal (n = 55) factors. In this study we evaluated FSH, estradiol, luteinizing hormone (LH), progesterone, prolactin and anti-Mullerian hormone levels, and antral follicle counting (AFC). Genotyping was performed using the TaqMan real-time polymerase chain reaction methodology. RESULTS: The wild-type allele G was found in 86.4% and the polymorphic allele T in 13.6% of the women respectively. The TT genotype was not found in any women. Women carrying the GT genotype had a poorer response more frequently to controlled ovarian hyperstimulation when compared to individuals with the GG genotype (47.4% vs. 26.5%, p = 0.010), higher LH levels (3.1 IU/mL vs. 2.4 IU/mL, p = <0.001), lower AFC (8.0 vs. 10.0, p = 0.03), oocytes retrieved (3.0 vs. 5.0, p = 0.03), MII (3.0 vs. 4.0, p = 0.02), and embryos (2.0 vs. 3.0, p = 0.02). Despite these findings, no difference was observed in pregnancy rate. CONCLUSION: Our findings suggest that the FSHB c.-211G/T polymorphism may modestly alter some aspects of the female reproductive system, but they are not associated with significantly different IVF outcomes.


Assuntos
Proteínas de Transporte/genética , Glicopeptídeos/genética , Adulto , Alelos , Hormônio Antimülleriano/genética , Brasil , Estudos Transversais , Feminino , Fertilização In Vitro , Hormônio Foliculoestimulante/genética , Hormônio Foliculoestimulante/metabolismo , Subunidade beta do Hormônio Folículoestimulante/genética , Frequência do Gene/genética , Genótipo , Humanos , Hormônio Luteinizante/genética , Polimorfismo de Nucleotídeo Único/genética , Gravidez , Taxa de Gravidez , Regiões Promotoras Genéticas/genética , Receptores do FSH , Saúde Reprodutiva
12.
PLoS One ; 13(11): e0207526, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30462715

RESUMO

Follicle-stimulating hormone receptor (FSHR) is a G-protein coupled receptor (GPCR) and a prototype of the glycoprotein hormone receptors subfamily of GPCRs. Structural data of the FSHR ectodomain in complex with follicle-stimulating hormone suggests a "pull and lift" activation mechanism that triggers a conformational change on the seven α-helix transmembrane domain (TMD). To analyze the conformational changes of the FSHR TMD resulting from sequence variants associated with reproductive impairment in humans, we set up a computational approach combining helix modeling and molecular simulation methods to generate conformational ensembles of the receptor at room (300 K) and physiological (310 K) temperatures. We examined the receptor dynamics in an explicit membrane environment of polyunsaturated phospholipids and solvent water molecules. The analysis of the conformational dynamics of the functional (N680 and S680) and dysfunctional (mutations at D408) variants of the FSHR allowed us to validate the FSHR-TMD model. Functional variants display a concerted motion of flexible intracellular regions at TMD helices 5 and 6. Disruption of side chain interactions and conformational dynamics were detected upon mutation at D408 when replaced with alanine, arginine, or tyrosine. Dynamical network analysis confirmed that TMD helices 2 and 5 may share communication pathways in the functional FSHR variants, whereas no connectivity was detected in the dysfunctional mutants, indicating that the global dynamics of the FSHR was sensitive to mutations at amino acid residue 408, a key position apparently linked to misfolding and variable cell surface plasma membrane expression of FSHRs with distinct mutations at this position.


Assuntos
Aminoácidos/química , Hormônio Foliculoestimulante/química , Conformação Molecular , Receptores do FSH/química , Alanina/química , Alanina/genética , Sequência de Aminoácidos/genética , Aminoácidos/genética , Simulação por Computador , Hormônio Foliculoestimulante/genética , Humanos , Bicamadas Lipídicas/química , Lipídeos/química , Lipídeos/genética , Simulação de Dinâmica Molecular , Mutação Puntual , Conformação Proteica em alfa-Hélice , Dobramento de Proteína , Receptores do FSH/genética , Transdução de Sinais
13.
Gen Comp Endocrinol ; 269: 149-155, 2018 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-30236970

RESUMO

In Seriola species, exposure to a long photoperiod regime is known to induce ovarian development. This study examined photoperiodic effects on pituitary gene expression and plasma levels of follicle-stimulating hormone (Fsh) and luteinizing hormone (Lh) in previtellogenic greater amberjack (Seriola dumerili). The fish were exposed to short (8L:16D) or long (18L:6D) photoperiod. The water temperature was maintained at 22 °C. Compared with the short-photoperiod group, plasma Fsh levels were higher on days 10 and 30 in the long-photoperiod group, but plasma Lh levels did not significantly differ. On day 30, pituitary Fsh- and Lh-ß subunit gene expressions were also higher in the long-photoperiod group than the short-photoperiod group, whereas α-subunit gene expressions were higher on days 20 and 30. Throughout the experiment, average gonadosomatic index and plasma E2 levels did not significantly differ between the two groups. This study clearly demonstrated that a long photoperiod induced Fsh release in the previtellogenic fish followed by upregulation of pituitary Fsh and Lh subunit gene expressions. An increase in plasma Fsh levels may be a key factor that mediates the photoperiodic effect on the initiation of ovarian development.


Assuntos
Gonadotropinas/sangue , Perciformes/sangue , Perciformes/fisiologia , Fotoperíodo , Vitelogênese , Animais , Estradiol/sangue , Feminino , Hormônio Foliculoestimulante/sangue , Hormônio Foliculoestimulante/genética , Hormônio Foliculoestimulante/metabolismo , Subunidade alfa de Hormônios Glicoproteicos/genética , Subunidade alfa de Hormônios Glicoproteicos/metabolismo , Hormônio Luteinizante/sangue , Hormônio Luteinizante/genética , Hormônio Luteinizante/metabolismo , Hormônio Luteinizante Subunidade beta/genética , Hormônio Luteinizante Subunidade beta/metabolismo , Ovário/crescimento & desenvolvimento , Perciformes/crescimento & desenvolvimento , Perciformes/metabolismo , Hipófise/citologia , Hipófise/metabolismo , Temperatura , Água
14.
Minerva Ginecol ; 70(5): 561-587, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30264954

RESUMO

This review article centers upon family of gonadotropin hormones which consists of two pituitary hormones - follicle-stimulating hormone (FSH) and luteinizing hormone (LH) as well as one non-pituitary hormone - human chorionic gonadotropin (hCG) secreted by placenta, and their receptors. Gonadotropins play an essential role in proper sexual development, puberty, gametogenesis, maintenance of pregnancy and male sexual differentiation during the fetal development. They belong to the family of glycoprotein hormones thus they constitute heterodimeric proteins built of common α subunit and hormone-specific ß-subunit. Hitherto, several mutations in genes encoding both gonadotropins and their receptors have been identified in humans. Their occurrence resulted in a number of different phenotypes including delayed puberty, primary amenorrhea, hermaphroditism, infertility and hypogonadism. In order to understand the effects of mutations on the phenotype observed in affected patients, detailed molecular studies are required to map the relationship between the structure and function of gonadotropins and their receptors. Nonetheless, in vitro assays are often insufficient to understand physiology. Therefore, several animal models have been developed to unravel the physiological roles of gonadotropins and their receptors.


Assuntos
Gonadotropina Coriônica/fisiologia , Hormônio Foliculoestimulante/fisiologia , Hormônio Luteinizante/fisiologia , Animais , Gonadotropina Coriônica/genética , Feminino , Doenças Urogenitais Femininas/genética , Doenças Urogenitais Femininas/fisiopatologia , Hormônio Foliculoestimulante/genética , Humanos , Hormônio Luteinizante/genética , Masculino , Doenças Urogenitais Masculinas/genética , Doenças Urogenitais Masculinas/fisiopatologia , Modelos Animais , Mutação , Fenótipo , Gravidez , Receptores da Gonadotropina/genética , Receptores da Gonadotropina/fisiologia
15.
PLoS One ; 13(7): e0197894, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30063719

RESUMO

BACKGROUND: Anti-Müllerian hormone (AMH) is expressed by granulosa cells of developing follicles and plays an inhibiting role in the cyclic process of follicular recruitment by determining follicle-stimulating hormone threshold levels. Knowledge of AMH expression in the porcine ovary is important to understand the reproductive efficiency in female pigs. RESEARCH AIM: In the present study we investigated the expression of AMH during follicular development in prepubertal and adult female pigs by immunohistochemistry, laser capture micro-dissection and RT-qPCR. RESULTS AND CONCLUSION: Although in many aspects the immunohistochemical localization of AMH in the porcine ovary does not differ from other species, there are also some striking differences. As in most species, AMH appears for the first time during porcine follicular development in the fusiform granulosa cells of recruited primordial follicles and continues to be present in granulosa cells up to the antral stage. By the time follicles reach the pre-ovulatory stage, AMH staining intensity increases significantly, and both protein and gene expression is not restricted to granulosa cells; theca cells now also express AMH. AMH continues to be expressed after ovulation in the luteal cells of the corpus luteum, a phenomenon unique to the porcine ovary. The physiological function of AMH in the corpus luteum is at present not clear. One can speculate that it may contribute to the regulation of the cyclic recruitment of small antral follicles. By avoiding premature exhaustion of the ovarian follicular reserve, AMH may contribute to optimization of reproductive performance in female pigs.


Assuntos
Hormônio Antimülleriano/genética , Corpo Lúteo/metabolismo , Hormônio Foliculoestimulante/genética , Aptidão Genética , Células da Granulosa/metabolismo , Células Tecais/metabolismo , Animais , Hormônio Antimülleriano/metabolismo , Corpo Lúteo/citologia , Feminino , Hormônio Foliculoestimulante/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Células da Granulosa/citologia , Imuno-Histoquímica , Ovulação/genética , Gravidez , Suínos , Células Tecais/citologia
16.
Appl Microbiol Biotechnol ; 102(20): 8867-8882, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30136206

RESUMO

Porcine follicle-stimulating hormone (pFSH), comprising α and ß subunits, is commonly used to induce superovulation in domestic animals in assisted reproduction technologies; however, the practical application of pFSH is inhibited by the limited efficiency of its production. Recombinant yeast-derived FSH offers a practical alternative; however, the heterologous expression efficiency remains disappointingly low. To improve FSH production in Pichia pastoris, a series of molecular strategies, together with fermentation optimization, were tested in the present study. By comparing clones of the Muts phenotype strain, it was observed that the yield of soluble pFSH increased by approximately 96% in clones of the Mut+ phenotype strain. The protein levels of soluble pFSHß, which confers biological specificity, increased by approximately 143 and 22% after two kinds of codon optimization strategies, respectively. Moreover, compared with the production of soluble pFSHß and SUMO-pFSHß, the production of soluble protein HSA-pFSHß was significantly improved. Furthermore, the optimum pH and methanol concentration for expressing soluble HSA-pFSH in strain H3-3 were determined as 5.0-6.0 and 1.5-2% in shake-flask, and the yield of soluble HSA-pFSH could reach 40.8 mg/l after purification. In vitro bioactivity assays showed that recombinant HSA-pFSH could efficiently stimulate cAMP synthesis in HEK293 cells expressing porcine FSHR. In conclusion, our results demonstrated that the application of phenotypic selection of aox1 mutants, combined with codon optimization, the choice of fusion partners, and fermentation optimization, considerably increased the yield of pFSH in supernatant of P. pastoris and thus provided a valuable reference for the large-scale recombinant expression of pFSH.


Assuntos
Hormônio Foliculoestimulante/genética , Expressão Gênica , Pichia/genética , Animais , Códon/genética , Códon/metabolismo , Meios de Cultura/química , Meios de Cultura/metabolismo , AMP Cíclico/metabolismo , Hormônio Foliculoestimulante/metabolismo , Hormônio Foliculoestimulante/farmacologia , Células HEK293 , Humanos , Pichia/metabolismo , Engenharia de Proteínas , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Suínos
17.
J Cell Biochem ; 119(12): 10131-10142, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30129147

RESUMO

Resveratrol (RSV), a polyphenolic compound largely found in red grape skin, has been used as a nutritional supplement as it exhibits beneficial health effects, such as anticancer, cardioprotective, antiaging, and anti-inflammatory. Particularly, it has been shown that it participates in the mechanisms involved in cell proliferation. Sirtuin 1 (SIRT1) is considered a well-known RSV effector. Noteworthy, Sirt1-knockout animals are infertile. The aim of this study was, first, to determine whether RSV has any effect on Sertoli cell proliferation and, second, whether SIRT1, a putative target of RSV, is present in immature Sertoli cells. Sertoli cell cultures obtained from 8-day-old rats, which actively proliferate, were treated with RSV (10 and 50 µM) under basal and follicle-stimulating hormone (FSH)-stimulated conditions. Bromodeoxyuridine (BrdU) incorporation and the expression of cyclins D1, D2, D3, E1, and E2 and the Cip/Kip cell cycle inhibitors p21Cip and p27Kip were analyzed. RSV decreased BrdU incorporation and cyclins D1, D2, E1, and E2 expression and increased p21Cip and p27Kip messenger RNA (mRNA) levels. RSV also decreased FSH-stimulated BrdU incorporation and cyclins D1 and D2 mRNA levels. The effect of RSV on cMYC was also analyzed. RSV treatment did not modify basal and FSH-stimulated cMyc expression; however, it inhibited basal and FSH-stimulated cMYC transcriptional activity, suggesting a role of cMYC in RSV effects. Additionally, Sirt1 was detected in immature Sertoli cells. Altogether, these results suggest that RSV possibly, by activating SIRT1 and regulating cMYC transcriptional activity, participates in the regulation of immature Sertoli cell proliferation.


Assuntos
Proliferação de Células/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-myc/genética , Resveratrol/farmacologia , Células de Sertoli/efeitos dos fármacos , Sirtuína 1/genética , Animais , Células Cultivadas , Hormônio Foliculoestimulante/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Proteínas de Neoplasias/genética , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Células de Sertoli/patologia
18.
J Assist Reprod Genet ; 35(9): 1703-1712, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29974367

RESUMO

PURPOSE: Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) mediate intracellular functions by binding their specific protein G-coupled gonadotrophin receptor, respectively FSH receptor (FSHR) and LH/choriogonadotrophin receptor (LHCGR). Whereas the expression of FSHR and LHCGR in mammals was considered gonad-specific and cell-specific, studies identified gonadotrophin receptors in human female extragonadal reproductive tissues. This study aims to demonstrate that gonadotrophin receptors are expressed in endometrium and mediates intracellular functions. METHODS: Collected endometria (n = 12) from healthy patients (mean age of 36 ± 6) were primary cultured for 24 h. The presence of gonadotrophin receptors was evaluated by RT-PCR followed by the sequencing of the resulted amplicons and by immunohistochemistry in original samples. Endometrial primary cultures were treated with increasing concentration (range 0-100 ng/ml) of either recombinant human LH (rhLH) or recombinant human FSH (rhFSH). Endometria controls had gonadotrophin replaced by the same volume of the culture medium. In gonadotrophin-treated samples, it was evaluated the intracellular cyclic adenosine monophosphate (cAMP) content by enzymatic immunoassay and the expression of steroidogenic genes by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR). RESULTS: The sequencing of the RT-PCR amplicons confirmed the presence of both gonadotrophin receptors and immunohistochemistry localized them on the membrane of endometrial glands cells throughout the glandular epithelium. The gonadotrophin-receptor complex was able to increase the intracellular cAMP in a dose-response and time-course manner and to induce steroidogenic genes expression. CONCLUSION: This study demonstrates that both gonadotrophin receptors are expressed along the glandular epithelium of endometria and they mediate the effects of gonadotrophins on intracellular functions.


Assuntos
Endométrio/metabolismo , Receptores do FSH/genética , Receptores do LH/genética , Adulto , Gonadotropina Coriônica/genética , AMP Cíclico/metabolismo , Endométrio/crescimento & desenvolvimento , Feminino , Hormônio Foliculoestimulante/genética , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Hormônio Luteinizante/genética , Hormônio Luteinizante/metabolismo , Gravidez , Receptores do FSH/metabolismo , Transdução de Sinais/genética
19.
Eur J Endocrinol ; 179(3): R143-R150, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29959220

RESUMO

The two pituitary gonadotrophins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), and in particular LH-stimulated high intratesticular testosterone (ITT) concentration, are considered crucial for spermatogenesis. We have revisited these concepts in genetically modified mice, one being the LH receptor (R)-knockout mouse (LuRKO), the other a transgenic mouse expressing in Sertoli cells a highly constitutively active mutated Fshr (Fshr-CAM). It was found that full spermatogenesis was induced by exogenous testosterone treatment in LuRKO mice at doses that restored ITT concentration to a level corresponding to the normal circulating testosterone level in WT mice, ≈5 nmol/L, which is 1.4% of the normal high ITT concentration. When hypogonadal LuRKO and Fshr-CAM mice were crossed, the double-mutant mice with strong FSH signaling, but minimal testosterone production, showed near-normal spermatogenesis, even when their residual androgen action was blocked with the strong antiandrogen flutamide. In conclusion, our findings challenge two dogmas of the hormonal regulation of male fertility: (1) high ITT concentration is not necessary for spermatogenesis and (2) strong FSH stimulation can maintain spermatogenesis without testosterone. These findings have clinical relevance for the development of hormonal male contraception and for the treatment of idiopathic oligozoospermia.


Assuntos
Hormônio Foliculoestimulante/fisiologia , Hormônio Luteinizante/fisiologia , Mutação , Espermatogênese/fisiologia , Animais , Cruzamentos Genéticos , Hormônio Foliculoestimulante/genética , Subunidade beta do Hormônio Folículoestimulante/genética , Humanos , Masculino , Camundongos , Camundongos Knockout , Camundongos Transgênicos , Receptores do FSH/genética , Receptores do LH/deficiência , Receptores do LH/genética , Células de Sertoli/metabolismo , Espermatogênese/efeitos dos fármacos , Espermatogênese/genética , Testículo/química , Testosterona/análise , Testosterona/farmacologia , Testosterona/fisiologia
20.
Genet Test Mol Biomarkers ; 22(7): 405-412, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29957069

RESUMO

AIMS: This is a follow-up study based on the results of our previous article, to further explore the effect of the TP53 codon 72 (rs1042522) and MDM2 SNP309 (rs2279744) polymorphisms on basal follicle-stimulating hormone (FSH)/luteinizing hormone (LH) ratios in infertility women. MATERIALS AND METHODS: The distribution of two genetic polymorphisms (rs1042522 and rs2279744) and basal FSH/LH ratios were tested and analyzed in 1051 in vitro fertilization (IVF) patients at a university-affiliated hospital. RESULTS: The TP53 codon 72 polymorphism had a significant association with the FSH/LH ratio (group I: FSH/LH <2.3 and group II: FSH/LH ≥2.3) (C/C vs. G/G: odds ratio [OR] = 1.69, 95% confidence interval [CI]: 1.07-2.65, p = 0.02; G/C vs. G/G: OR = 1.86, 95% CI: 1.25-2.77, p = 0.002). In a stratification analysis, C allele carriers and the C/C genotype showed a strong association with positive clinical pregnancy outcomes after IVF compared with G allele carriers and the G/G genotype in the recessive, dominant, and allelic genetic models in group I (C/C vs. G/G: OR = 1.84, 95% CI: 1.25-2.69, p = 0.01; C/C vs. G carrier: OR = 1.52, 95% CI: 1.12-2.07, p = 0.01; C carrier vs. G/G: OR = 1.46, 95% CI: 1.07-2.01, p = 0.02; C allele vs. G allele: OR = 1.34, 95% CI: 1.11-1.62, p = 0.003), no significant associations by stratification were observed for group II. No associations were found between MDM2 SNP309 and either of two groups. CONCLUSION: The TP53 codon 72 polymorphism is associated with FSH/LH ratios, suggesting that it is a potential predictive genetic marker of IVF outcome in patients younger than 35 years of age with baseline FSH levels below 10 IU/L and who have an FSH/LH ratio <2.3.


Assuntos
Hormônio Foliculoestimulante/metabolismo , Infertilidade Feminina/genética , Infertilidade Feminina/metabolismo , Hormônio Luteinizante/metabolismo , Proteínas Proto-Oncogênicas c-mdm2/genética , Proteína Supressora de Tumor p53/genética , Adulto , Grupo com Ancestrais do Continente Asiático/genética , Biomarcadores/análise , China , Códon , Feminino , Hormônio Foliculoestimulante/genética , Seguimentos , Frequência do Gene , Genótipo , Humanos , Hormônio Luteinizante/genética , Indução da Ovulação , Polimorfismo de Nucleotídeo Único , Proteínas Proto-Oncogênicas c-mdm2/metabolismo , Proteína Supressora de Tumor p53/metabolismo
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