Project description:Background: Melatonin (MLT) supplementation has been shown to improve the quality of oocytes and embryos in vitro through its antioxidant mechanism. However, it is unclear whether endogenous MLT plays a role in oocyte quality control, casting doubt on the rationality of oral MLT in improving oocyte quality or fecundity. Methods:we measured the effects of PMSG on MLT synthesis in follicles, evaluated the effects of MLT on oocyte growth and quality in mice and goat, used RNA-seq to explore the mechanism underlying MLT’s effects on oocyte growth and quality, and generated SNAT-knockout mice to assess the effects of endogenous MLT on oocyte growth and quality. Lastly, we investigated the feasibility of short-term MLT intake to improve the fertility of older mice. Result: we revealed that the follicle-stimulating hormone induces the expression of intra-ovarian SNAT, resulting in increased levels of intra-ovarian MLT in mice. Administering MLT during the gonadotropin-dependent phase improved the oocyte size and quality in both mice and goat by enhancing nutrient accumulation and mitochondrial function. Deficiency of MLT in mice reduced oocyte size and weakened mitochondrial function. Furthermore, our findings demonstrate that short-term MLT intake during the gonadotropin phase improves the litter size of older mice. Conclusion: This study reveals that endogenous MLT is involved in determining oocyte quality as a target molecule of follicle-stimulating hormone, shedding light on its potential utilization in improving oocyte quality or rejuvenating aged mammalian oocytes.
Project description:Follicle-stimulating hormone (FSH) is crucial in the development and regulation of reproductive functions. We expressed MAC-tagged FSH receptor (FSHR) in HEK293 cells followed by affinity purification mass spectrometry analyses, to investigate interactome of FSHR at resting state and upon FSH stimulation. In total, 19 specific high-confidence interactors for WT FSHR and 14 for A189V FSHR, several of which have been linked to infertility.
Project description:Differences in follicle-stimulating hormone (FSH) ß-subunit N-glycosylation results in distinct FSH glycoforms. Hypoglycosylated FSH21 is more bioactive and abundant in reproductively young women, whereas fully glycosylated FSH24 shows lower bioactivity and a relative increase with age. To investigate if the coinciding shift in FSH glycoform abundance contributes to the age-dependent decline in oocyte quality, an encapsulated in vitro follicle growth system was utilized to examine the direct effects of FSH glycoforms on folliculogenesis and resulting oocyte quality. Long-term culture (10-12 days) with FSH21 (10 ng/ml) enhanced follicle growth, estradiol secretion, and oocyte quality compared to equivalent FSH24 treatment. FSH21 exhibited enhanced capacity to establish transzonal projections, gap junctions, and cell-to-cell communication within 24 hrs in culture. Transient inhibition of FSH21-mediated bidirectional communication abrogated the positive effects of FSH21 on follicle growth, estradiol secretion and oocyte quality. These findings indicate FSH21 promotes folliculogenesis and oocyte quality in vitro through increasing cell-to-cell communication early in folliculogenesis and that the shift in abundance from FSH21 to FSH24 with reproductive aging could contribute to the age-dependent decline in oocyte quality.
Project description:FSH is considered as the most critical regulatory factor for follicle growth and development. It is widely acknowledged that exogenous administration of FSH can effectively stimulate ovulation.However, these techniques have several drawbacks such as high cost and detrimental effects on hormone balance and embryo implantation.Sulforaphane (SFN) is a naturally occurring compound that belongs to the isothiocyanate group, which is predominant in cruciferous vegetables.Our previous research demonstrated SFN could inhibit hypoxia-evoked apoptosis in porcine GCs. Intriguingly, SFN exhibits a biphasic dose-response, stimulating cell growth at low doses in various mammalian cells. Therefore,to investigate the functional implications of sulforaphane in the regulation of granulosa cell proliferation, we conducted a gene expression profiling analysis using RNA-seq data, comparing SFN to follicle stimulating hormone (FSH).
Project description:Follicle-stimulating hormone (FSH), a dimeric glycoprotein produced by pituitary gonadotrope cells, regulates spermatogenesis in males and ovarian follicle growth in females. Hypothalamic gonadotropin-releasing hormone (GnRH) stimulates FSHβ subunit gene (Fshb) transcription, though the underlying mechanisms are poorly understood. To address this gap in knowledge, we examined changes in pituitary gene expression in GnRH-deficient mice (hpg) treated with a regimen of exogenous GnRH that increases pituitary Fshb but not luteinizing hormone β (Lhb) mRNA levels. Activating transcription factor 3 (Atf3) was among the most upregulated genes. ATF3 can heterodimerize with members of the AP-1 family to regulate gene transcription. Co-expression of ATF3 with JunB stimulated murine Fshb, but not Lhb, promoter-reporter activity in homologous LβT2 cells. ATF3 also synergized with a constitutively active activin type I receptor to increase endogenous Fshb expression in these cells. Nevertheless, FSH production was intact in gonadotrope-specific Atf3 knockout mice (cKO) and control littermates. Ovarian follicle development, ovulation, and litter sizes were also equivalent between genotypes. Testis weights and sperm counts did not differ between cKO and control males. Following gonadectomy, increases in LH secretion were enhanced in cKO animals. Though FSH levels did not differ between genotypes, post-gonadectomy increases in pituitary Fshb and gonadotropin α subunit expression were more pronounced in cKO mice. These data indicate that ATF3 can selectively stimulate Fshb transcription in vitro but is not required for FSH production in vivo.