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Journal of Lipid Research, Vol. 50, 2164-2172, November 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

Reducing CYP51 inhibits follicle-stimulating hormone induced resumption of mouse oocyte meiosis in vitro

Chao Wang^1,*, Baoshan Xu^1,, Bo Zhou, Cheng Zhang, Jie Yang, Hong Ouyang, Gang Ning, Meijia Zhang, Jianzhong Shen and Guoliang Xia^2,

^* School of Basic Medical Sciences, Capital Medical University, Beijing 100069, PR China
State Key Laboratories for AgroBiotechnology and Department of Animal Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing 100094, PR China
College of Veterinary Medicine, China Agricultural University, Beijing 100094, PR China

² To whom correspondence should be addressed. e-mail: glxiachina{at}sohu.com

Meiosis activating sterol, produced directly by lanosterol 14--demethylase (CYP51) during cholesterol biosynthesis, has been shown to promote the initiation of oocyte meiosis. However, the physiological significance of CYP51 action on oocyte meiosis in response to gonadotrophins’ induction remained to be further explored. Herein, we analyzed the role of CYP51 in gonadotrophin-induced in vitro oocyte maturation via RNA interference (RNAi). We showed that although both luteinizing hormone (LH) and follicle-stimulating hormone (FSH) significantly induced meiotic resumption in follicle-enclosed oocytes (FEOs), the effect of LH on oocyte meiosis resumption in FEOs was weaker than FSH. Moreover, both FSH and LH were able to upregulate CYP51 expression in cultured follicular granulosa cells when examined at 8 h or 12 h posttreatments, respectively. Interestingly, whereas knockdown of CYP51 expression via small interference RNA (siRNA) moderately blocked (23% reduction at 24 h) FSH-induced oocyte maturation [43% germinal vesicle breakdown (GVBD) rate in RNAi vs. 66% in control, P < 0.05] in FEOs, similar treatments showed no apparent effects on LH-induced FEO meiotic maturation (58% GVBD rate in RNAi vs. 63% in control, P > 0.05). Moreover, the results in a cumulus-enclosed oocytes (CEOs) model showed that approximately 30% of FSH-induced CEOs’ meiotic resumption was blocked upon CYP51 knockdown by siRNAs. These findings suggest that FSH, partially at least, employs CYP51, and therefore the MAS pathway, to initiate oocyte meiosis.

Supplementary key words RNA interference • meiotic resumption • oocyte

Abbreviations: CEO, cumulus cells-enclosed oocyte; CYP51, lanosterol 14--demethylase; CYP-mis, CYP51-mismatched siRNA; eCG, equine chorionic gonadotrophin; EGFP, enhance green florescence protein plasmid; FEO, follicle-enclosed oocyte; FSH, follicle-stimulating hormone; GVBD, germinal vesicle breakdown; hCG, human chorionic gonadotrophin; HX, hypoxanthine; LH, luteinizing hormone; MAS, meiosis activating sterol; RNAi, RNA interference; siRNA, small interference RNA; TBST, TBS containing 0.1% Tween-20

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