The luteinizing hormone receptor (LHCGR) has a little studied polymorphic 6 bp insertion (rs4539842/insLQ). This study has evaluated the insLQ polymorphism in relation to potential associations with hormonal characteristics of human small antral follicles (hSAFs). In total, 310 hSAFs were collected from 86 women undergoing fertility preservation. Analysis included hormonal profile of 297 follicular fluid (FF) samples and 148 corresponding granulosa cells samples were evaluated by qPCR for selected genes. Significantly reduced and non-detectable mRNA levels of anti-Müllerian hormone receptor II (AMHR2) and LHCGR, respectively, were observed for insLQ/insLQ compared to -/insLQ and the -/- genotypes. Moreover, LHCGR and CYP19a1 together with oestradiol and inhibin-B were significantly increased in -/insLQ compared to the -/- genotype. The homozygous insLQ genotype showed strong significant associations to GC specific genes LHCGR and CYP19a1, which may translate into significant changes in FF hormone profiles and an altered LH signaling.
- Klíčová slova
- Follicle fluid, Human small antral follicles, LHR, rs4539842,
- MeSH
- dospělí MeSH
- folikulární buňky metabolismus MeSH
- folikulární tekutina metabolismus MeSH
- genotyp MeSH
- hormony metabolismus MeSH
- inzerční mutageneze * MeSH
- lidé MeSH
- messenger RNA genetika metabolismus MeSH
- mladiství MeSH
- mladý dospělý MeSH
- ovariální folikul metabolismus MeSH
- polymorfismus genetický * MeSH
- receptory LH genetika MeSH
- regulace genové exprese * MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- hormony MeSH
- messenger RNA MeSH
- receptory LH MeSH
The key mechanisms responsible for achievement of full reproductive and developmental capability in mammals are the differentiation and transformation of granulosa cells (GCs) during folliculogenesis, oogenesis, and oocyte maturation. Although the role of 17 beta-estradiol (E2) in ovarian activity is widely known, its effect on proliferative capacity, gap junction connection (GJC) formation, and GCs-luteal cells transformation requires further research. Therefore, the goal of this study was to assess the real-time proliferative activity of porcine GCs in vitro in relation to connexin (Cx), luteinizing hormone receptor (LHR), follicle stimulating hormone receptor (FSHR), and aromatase (CYP19A1) expression during short-term (168 h) primary culture. The cultured GCs were exposed to acute (at 96 h of culture) and/or prolonged (between 0 and 168 h of culture) administration of 1.8 and 3.6 μM E2. The relative abundance of Cx36, Cx37, Cx40, Cx43, LHR, FSHR, and CYP19A1 mRNA was measured. We conclude that the proliferation capability of GCs in vitro is substantially associated with expression of Cxs, LHR, FSHR, and CYP19A1. Furthermore, the GC-luteal cell transformation in vitro may be significantly accompanied by the proliferative activity of GCs in pigs.
- MeSH
- buněčná diferenciace účinky léků genetika MeSH
- estradiol aplikace a dávkování MeSH
- folikulární buňky účinky léků metabolismus MeSH
- IVM techniky MeSH
- lidé MeSH
- oocyty účinky léků růst a vývoj MeSH
- oogeneze účinky léků genetika MeSH
- prasata MeSH
- proliferace buněk účinky léků MeSH
- receptory FSH biosyntéza genetika MeSH
- receptory LH biosyntéza genetika MeSH
- rodina 19 cytochromů P450 biosyntéza MeSH
- vývojová regulace genové exprese účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- estradiol MeSH
- receptory FSH MeSH
- receptory LH MeSH
- rodina 19 cytochromů P450 MeSH
Vinclozolin (VZ) is a pesticide that acts as an anti-androgen to impair reproduction in mammals. However, VZ-induced disruption of reproduction is largely unknown in fish. In the present study, we have established a combination exposure in which adult goldfish were exposed to VZ (30 and 100 μg/L), anti-androgen flutamide (Flu, 300 μg/L), and androgen testosterone (T, 1 μg/L) to better understand effects of VZ on reproductive endocrine system. mRNA levels of kisspeptin (kiss-1 and kiss-2) and its receptor (gpr54), salmon gonadotropin-releasing hormone (gnrh3) and androgen receptor (ar) in the mid-brain, and luteinizing hormone receptor (lhr) in the testis were analyzed and compared with those of control following 10 days of exposure. kiss-1 mRNA level was increased in goldfish exposed to 100 µg/L VZ and to Flu, while kiss-2 mRNA level was increased following exposure to Flu and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. gpr54 mRNA level was increased in goldfish exposed to Flu and to combination of 30 µg/L VZ with Flu and 100 µg/L VZ with T. gnrh3 mRNA level was increased in goldfish exposed to 100 µg/L VZ, to Flu, and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. The mid-brain ar mRNA level was increased in goldfish exposed to Flu and to combinations of 30 µg/L VZ with Flu, 100 µg/L VZ with T, and Flu with T. Testicular lhr mRNA level was increased in goldfish exposed to Flu and to combination of 30 µg/L VZ with Flu. These results suggest that VZ and Flu are capable of interfering with kisspeptin and GnRH systems to alter pituitary and testicular horonal functions in adult goldfish and the brain ar mediates VZ-induced disruption of androgen production.
- Klíčová slova
- Androgen receptor, Flutamide, Kisspeptin, Luteinizing hormone receptor, Salmon gonadotropin-releasing hormone, Testosterone,
- MeSH
- androgenní receptory genetika MeSH
- androgeny farmakologie MeSH
- antagonisté androgenů farmakologie MeSH
- flutamid farmakologie MeSH
- hormon uvolňující gonadotropiny genetika MeSH
- karas zlatý genetika MeSH
- kisspeptiny genetika MeSH
- kyselina pyrrolidonkarboxylová analogy a deriváty MeSH
- lékové interakce MeSH
- messenger RNA metabolismus MeSH
- mozek účinky léků metabolismus MeSH
- oxazoly farmakologie MeSH
- průmyslové fungicidy farmakologie MeSH
- receptory LH genetika MeSH
- receptory spřažené s G-proteiny genetika MeSH
- rozmnožování účinky léků MeSH
- rybí proteiny genetika MeSH
- testis účinky léků metabolismus MeSH
- testosteron farmakologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- androgenní receptory MeSH
- androgeny MeSH
- antagonisté androgenů MeSH
- flutamid MeSH
- gonadotropin-releasing hormone-III MeSH Prohlížeč
- hormon uvolňující gonadotropiny MeSH
- kisspeptiny MeSH
- kyselina pyrrolidonkarboxylová MeSH
- messenger RNA MeSH
- oxazoly MeSH
- průmyslové fungicidy MeSH
- receptory LH MeSH
- receptory spřažené s G-proteiny MeSH
- rybí proteiny MeSH
- testosteron MeSH
- vinclozolin MeSH Prohlížeč
Granulosa cell (GC) expressed androgen receptors (AR) and intrafollicular androgens are central to fertility. The transactivating domain of the AR contains a polymorphic CAG repeat sequence, which is linked to the transcriptional activity of AR and may influence the GC function. This study aims to evaluate the effects of the AR CAG repeat length on the intrafollicular hormone profiles, and the gene expression profiles of GC from human small antral follicles. In total, 190 small antral follicles (3-11 mm in diameter) were collected from 58 women undergoing ovarian cryopreservation for fertility preservation. The biallelic mean of the CAG repeat lengths were calculated for each woman, and grouped in three groups: Long CAG repeats (23-26 mean CAG); medium CAG repeats (20.5-22.5 mean CAG) and short CAG repeats (17.5-20.0 mean CAG). The following parameters were measured: follicle diameter, intrafollicular levels of Anti-Müllerian Hormone (AMH), progesterone, oestradiol, testosterone and androstenedione, and GC gene expression levels of FSHR, LHR, AR, CYP19A1, and AMH. The long CAG repeat lengths were associated with significantly decreased testosterone levels, as compared to medium CAG repeats (P = 0.05) and short CAG repeats (P = 0.003). Furthermore, in follicles 3-6 mm in diameter, the long CAG repeats were associated with significantly increased LHR and CYP19A1 gene expression levels compared to short CAG repeat lengths (P = 0.004 and P = 0.04 respectively), and significantly increased LHR expression compared to medium CAG repeat lengths (P = 0.03). In conclusion, long CAG repeat lengths in the AR were associated to significant attenuated levels of androgens and an increased conversion of testosterone into oestradiol, in human small antral follicles.
- Klíčová slova
- Androgen levels, Androgen receptor, CAG repeats, Follicular fluid, Human small antral follicles,
- MeSH
- androgenní receptory genetika MeSH
- aromatasa genetika MeSH
- dospělí MeSH
- expanze trinukleotidových repetic * MeSH
- folikulární tekutina cytologie metabolismus MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- pohlavní hormony genetika metabolismus MeSH
- receptory LH genetika MeSH
- stanovení celkové genové exprese metody MeSH
- testosteron metabolismus MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- androgenní receptory MeSH
- AR protein, human MeSH Prohlížeč
- aromatasa MeSH
- CYP19A1 protein, human MeSH Prohlížeč
- pohlavní hormony MeSH
- receptory LH MeSH
- testosteron MeSH
We show in the present study that freshly isolated pig cumulus-oocyte complexes (COCs) display a limited response to LH, as assessed by the expression of hyaluronan synthase 2 (Has2) mRNA, activation of protein kinase A (PKA), production of hyaluronic acid (HA) and progesterone, cumulus cell expansion and resumption of meiosis. These data indicate that freshly isolated COCs do not possess a sufficient number of functional LH receptors (LHR). However, the expression of Lhr significantly increased during the culture of COCs in vitro in a medium supplemented with FSH. Assuming that the effect of FSH on LHR induction is mediated via cAMP signaling pathways, we developed a new culture system, in which the COCs were pre-cultured for 72 hr in a medium supplemented with dbcAMP. The pre-cultured COCs remained in the germinal vesicle stage, their cumulus investment underwent a dramatic increase in size and gap junctions between the cumulus cells were preserved. The stimulation of such COCs with either FSH or LH led to the resumption and completion of meiosis, activation of PKA, expression of Has2, synthesis of large amounts of HA and progesterone, and extensive expansion of cumulus cells. We conclude that the formation of functional LHR is stimulated in cumulus cells during the culture in vitro in a cAMP-dependent pathway. The dbcAMP-treated COCs thus represent a new model in which the resumption of meiosis and cumulus expansion can be induced exclusively by the action of recombinant LH.
- MeSH
- analýza rozptylu MeSH
- buněčné kultury metody MeSH
- dibutyryl cyklický AMP farmakologie MeSH
- exprese genu účinky léků MeSH
- folikuly stimulující hormon farmakologie MeSH
- glukuronosyltransferasa biosyntéza genetika metabolismus MeSH
- kumulární buňky cytologie účinky léků metabolismus MeSH
- kyselina hyaluronová metabolismus MeSH
- luteinizační hormon farmakologie MeSH
- messenger RNA biosyntéza genetika MeSH
- oocyty cytologie účinky léků růst a vývoj MeSH
- ovariální folikul účinky léků metabolismus MeSH
- prasata MeSH
- progesteron metabolismus MeSH
- proteinkinasy závislé na cyklickém AMP metabolismus MeSH
- receptory LH biosyntéza genetika metabolismus MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- dibutyryl cyklický AMP MeSH
- folikuly stimulující hormon MeSH
- glukuronosyltransferasa MeSH
- kyselina hyaluronová MeSH
- luteinizační hormon MeSH
- messenger RNA MeSH
- progesteron MeSH
- proteinkinasy závislé na cyklickém AMP MeSH
- receptory LH MeSH
