Tetsuka Masafumi, Nishimoto Hiromi, Miyamoto Akio, Okuda Kiyoshi, Hamano Seizo
Department of Agricultural and Life Science, Obihiro University of Agriculture & Veterinary Medicine, Hokkaido, Japan.
J Reprod Dev. 2010 Dec;56(6):616-22. doi: 10.1262/jrd.10-019k. Epub 2010 Aug 11.
Glucocorticoids modulate ovarian function in cattle. However, their regulatory mechanisms have not been fully elucidated. In the present study, we examined gene expression of two glucocorticoid-metabolizing enzymes, a bidirectional 11β-HSD type 1 (11HSD1) and a dehydrogenase 11β-HSD type 2 (11HSD2), and glucocorticoid receptor (GR) in bovine follicles during follicular maturation and atresia. Granulosa cells (GCs) and theca interna layers (TIs) were harvested from follicles classified as small growing, dominant, preovulatory, early atretic and late atretic follicles. The expression levels of 11HSD1, 11HSD2 and GR mRNA were quantified by real-time PCR. In the healthy follicles, expression of 11HSD1 mRNA increased as follicles matured, both in GCs and TIs. A significant negative correlation was found between the concentration of cortisol in follicular fluid and the level of 11HSD1 mRNA in GCs. The expression of 11HSD2 and GR was either very low or largely unchanged during follicular maturation. In the atretic follicles, a drastic increase in the expression of 11HSD2 was observed both in GCs and TIs. To assess the effect of FSH on the expression of 11HSDs and GR, GCs were cultured with FSH (0-100 ng/ml) for up to 6 days. FSH increased 11HSD1 mRNA in a dose-dependent manner, but not 11HSD2, nor GR. Taken together, these results suggest that developmentally-regulated 11HSD1 plays a pivotal role in modulating the local glucocorticoid environment in maturing bovine follicles.
糖皮质激素可调节母牛的卵巢功能。然而,其调控机制尚未完全阐明。在本研究中,我们检测了双向11β-羟基类固醇脱氢酶1型(11HSD1)和脱氢酶11β-羟基类固醇脱氢酶2型(11HSD2)这两种糖皮质激素代谢酶以及糖皮质激素受体(GR)在牛卵泡发育成熟和闭锁过程中的基因表达。从分类为小生长卵泡、优势卵泡、排卵前卵泡、早期闭锁卵泡和晚期闭锁卵泡的卵泡中收集颗粒细胞(GCs)和卵泡内膜层(TIs)。通过实时PCR定量11HSD1、11HSD2和GR mRNA的表达水平。在健康卵泡中,随着卵泡成熟,GCs和TIs中11HSD1 mRNA的表达均增加。卵泡液中皮质醇浓度与GCs中11HSD1 mRNA水平之间存在显著负相关。在卵泡成熟过程中,11HSD2和GR的表达要么非常低,要么基本不变。在闭锁卵泡中,GCs和TIs中均观察到11HSD2的表达急剧增加。为了评估促卵泡素(FSH)对11HSDs和GR表达的影响,将GCs与FSH(0 - 100 ng/ml)培养长达6天。FSH以剂量依赖性方式增加11HSD1 mRNA,但不增加11HSD2和GR。综上所述,这些结果表明发育调控的11HSD1在调节成熟牛卵泡中的局部糖皮质激素环境中起关键作用。