In Vitro Culture of Fetal Ovaries: A Model to Study Factors Regulating Early Follicular Development

Roy, Shyamal K.; Wang, Cheng; Mukherjee, Anindit; Chakraborty, Prabuddha

doi:10.1007/978-1-61779-436-0_12

Cited by 7 publications

(4 citation statements)

References 25 publications

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“…Although somatic cell proliferation is expected to contribute to higher FSHR mRNA levels as the ovary grows during the postnatal life, it is logical to speculate that the processing of FSHR mRNA is stabilized as serum FSH levels increase (Roy and Hughes, 1994). The higher levels of FSHR mRNA preceding the day of primordial follicle development (P8) support our hypothesis that FSH affects ovarian somatic cell differentiation into pregranulosa cells (Roy et al, 2012, Wang and Roy, 2009, Wang and Roy, 2004). …”

Section: Discussionsupporting

confidence: 76%

“…FSH deprivation with an anti-FSH antiserum during fetal ovary development compromises primordial follicle formation in P8 hamsters (Roy and Albee, 2000). That the FSHR is functional is evident from our previous data, which show increased primordial follicle formation and cAMP production by postnatal hamster ovaries in response to FSH (Roy and Albee, 2000, Roy, Wang, Mukherjee et al, 2012, Wang and Roy, 2004). The distinct FSHR immunosignal in antral granulosa cells of follicles, but not in cells of the theca, liver or muscle, and immunoblot signal in proteins of antral granulosa cells and KGN cells, but not of the muscle, liver or uterus indicates the specificity of the FSHR antibody, although the limited potency presents a limitation for low amount of sample protein.…”

Section: Discussionmentioning

confidence: 67%

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Expression of FSH receptor in the hamster ovary during perinatal development

Chakraborty

Roy

2015

Molecular and Cellular Endocrinology

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FSH plays an important role in ovarian follicular development, and it functions via the G-protein coupled FSH receptor. The objectives of the present study were to determine if full-length FSHR mRNA and corresponding protein were expressed in fetal through postnatal hamster ovaries to explain the FSH-induced primordial follicle formation, and if FSH or estrogen (E) would affect the expression. A full-length and two alternately spliced FSHR transcripts were expressed from E14 through P20. The level of the full-length FSHR mRNA increased markedly through P7 before stabilizing at a lower level with the formation and activation of primordial follicles. A predicted 87kDa FSHR protein band was detected in fetal through P4 ovaries, but additional bands appeared as ovary developed. FSHR immunosignal was present in undifferentiated somatic cells and oocytes in early postnatal ovaries, but was granulosa cells specific after follicles formed. Both eCG and E significantly up-regulated full-length FSHR mRNA levels. Therefore, FSHR is expressed in the hamster ovary from the fetal life to account for FSH-induced primordial follicle formation and cAMP production. Further, FSH or E regulates the receptor expression.

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Section: Discussionsupporting

confidence: 76%

Section: Discussionmentioning

confidence: 67%

Expression of FSH receptor in the hamster ovary during perinatal development

Chakraborty

Roy

2015

Molecular and Cellular Endocrinology

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“…Ovaries were collected between 0800 and 1000 hours from embryonic day 15 (E15) hamsters in phenol red-free DMEM containing antibiotics at room temperature, cleaned of adherent tissues, and cultured in the presence of 0.1 μg/ml insulin, and 1.25 μg/ml transferrin, 1.25 μg/ml selenium, and 10.7 μg/ml linoleic acid (ITS; as described previously) 44 45 with or without either 50 ng/ml of BMP2 (R & D Systems, Minneapolis, MN) or 500 nM LDN 193,189 (ALK2/3 inhibitor) or 200 nM DMH1 (ALK2 inhibitor). All ovaries were placed in culture by 1300 hours of the day of collection.…”

Section: Methodsmentioning

confidence: 99%

Bone morphogenetic protein 2 promotes primordial follicle formation in the ovary

Chakraborty

Roy

2015

Sci Rep

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Primordial follicles (PF) are formed when somatic cells differentiate into flattened pregranulosa cells, invaginate into the oocyte nests and encircle individual oocytes. We hypothesize that BMP2 regulates PF formation by promoting the transition of germ cells into oocytes and somatic cells into pregranulosa cells. E15 hamster ovaries were cultured for 8 days corresponding to postnatal day 8 (P8) in vivo, with or without BMP2, and the formation of PF was examined. BMP2 was expressed in the oocytes as well as ovarian somatic cells during development. BMP2 exposure for the first two days or the last two days or the entire 8 days of culture led to increase in PF formation suggesting that BMP2 affected both germ cell transition and somatic cell differentiation. Whereas an ALK2/3 inhibitor completely blocked BMP2-induced PF formation, an ALK2-specific inhibitor was partially effective, suggesting that BMP2 affected PF formation via both ALK2 and ALK3. BMP2 also reduced apoptosis in vitro. Further, more meiotic oocytes were present in BMP2 exposed ovaries. In summary, the results provide the first evidence that BMP2 regulates primordial follicle formation by promoting germ cell to oocyte transition and somatic cell to pre-granulosa cells formation and it acts via both ALK2 and ALK3.

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“…The embedded ovarian tissue was sliced into 6 µm sections and stained with hematoxylin and eosin. Follicles were classified and counted under 400× magnification with an AXIO Imager A1 (Carl Zeiss Microscopy Co., Ltd.) as reported previously (Roy et al 2012). We defined primordial follicles as those containing flat GCs, primary follicles as those with a single layer of cuboidal GCs, secondary follicles as those having two to five layers of cuboidal GCs and antral follicles as those surrounded by more than five layers of GCs forming the antral cavity.…”

Section: Murine Ovarian Tissue Culture and Follicle Dynamic Analysismentioning

confidence: 99%

Effect of the neuropeptide phoenixin and its receptor GPR173 during folliculogenesis

Nguyen¹,

Nakamura²,

Osuka³

et al. 2019

Reproduction

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Folliculogenesis is a complex process, defined by the growth and development of follicles from the primordial population. Granulosa cells (GCs) play a vital role in every stage of follicular growth through proliferation, acquisition of gonadotropic responsiveness, steroidogenesis and production of autocrine/paracrine factors. A recently discovered hypothalamic neuropeptide phoenixin is involved in the regulation of the reproductive system. Phoenixin acts through its receptor, G protein-coupled receptor 173 (GPR173), to activate the cAMP/PKA pathway leading to the phosphorylation of CREB (pCREB). Here, we demonstrated the expression patterns of phoenixin and GPR173 in human ovary and explored its role in folliculogenesis. Phoenixin and GPR173 were both expressed in the human ovarian follicle, with increased expression in GCs as the follicle grows. Phoenixin treatment at 100 nM for 24 h induced the proliferation of human non-luteinized granulosa cell line, HGrC1 and significantly increased the expression levels of CYP19A1, FSHR, LHR and KITL, but decreased NPPC expression levels. These effects were suppressed by GPR173 siRNA. The expression level of CREB1, pCREB and estradiol (E2) production in the culture medium was significantly enhanced by phoenixin treatment in a concentration-dependent manner. Phoenixin also significantly increased the follicular area in a murine ovarian tissue culture model, leading to an increased number of ovulated oocytes with a higher level of maturation. Taken together, our data demonstrate that phoenixin is an intraovarian factor that promotes follicular growth through its receptor GPR173 by accelerating proliferation of GCs, inducing E2 production and increasing the expression of genes related to follicle development.

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In Vitro Culture of Fetal Ovaries: A Model to Study Factors Regulating Early Follicular Development

Cited by 7 publications

References 25 publications

Expression of FSH receptor in the hamster ovary during perinatal development

Expression of FSH receptor in the hamster ovary during perinatal development

Bone morphogenetic protein 2 promotes primordial follicle formation in the ovary

Effect of the neuropeptide phoenixin and its receptor GPR173 during folliculogenesis

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