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Expression of growth differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15) and anti-mullerian hormone (AMH) in cultured mouse primary follicles

J Sadeu, Follicle Biology Laboratory, Dutch-speaking Free University of Brussels (Vrije Universiteit Brussel), Brussels, 1090, Belgium
T Adriaenssens, Dutch-speaking Free University of Brussels (Vrije Universiteit Brussel), Follicle Biology Laboratory, Brussels -Belgium-, Belgium
J Smitz, Dutch-speaking Free University of Brussels (Vrije Universiteit Brussel), Follicle Biology Laboratory, Brussels -Belgium-, Belgium

Correspondence: Jean Clair Sadeu, Email: vsadeu{at}yahoo.co.uk

Abstract

Growth and differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15) and anti-mullerian hormone (AMH) play an important role in the primary-to-secondary follicle transition and follicle activation in vivo. In organ culture of neonatal mouse ovaries, it was observed that significantly less primary follicles develop to the secondary stage. The objectives of this study were: 1) to compare ovarian follicular populations between organ-cultured neonatal mouse ovaries and freshly isolated age-matched control ovaries; 2) to quantify RNA levels of GDF-9, BMP-15 and AMH, in cultured primary follicles; and 3) to immunolocalize GDF-9 and AMH in cultured ovaries. Ovaries from 3-day-old (PND 3) mice were cultured for 7 or 10 days in absence or presence of FSH. Follicular populations were counted in freshly isolated 13-day-old (PND 13) ovaries and organ-cultured ovaries. Transcripts were quantified in isolated primary follicles using real-time RT-PCR, and protein expressions were localized using immunohistochemistry. The number of secondary follicles in organ cultured ovaries was significantly lower than in vivo, controls. GDF-9 and BMP-15 mRNA expression levels were similar as in controls. AMH mRNA levels were significantly (P < 0.05) lower after day 10 of culture in absence of FSH. GDF-9 and AMH proteins were respectively detected in the oocytes and granulosa cells (GC) beginning at the primary and primordial stage onward. GDF-9 and BMP-15 production in cultured primary follicles are not different from in vivo controls; hence abnormal early follicle growth was not related to a deficient transcription of these factors.