Zhang Yi-Ming, Roy Shyamal K
Department of Physiology and Biophysics, University of Nebraska Medical Center, Omaha, 68198-4515, USA.
Biol Reprod. 2004 Jun;70(6):1580-8. doi: 10.1095/biolreprod.103.026898. Epub 2004 Jan 28.
Although gonadotropins have been reported to downregulate FSH-receptor (FSHR) mRNA levels in the ovaries of female rats, the effect of the gonadotropin surge, particularly FSH, on hamster follicular FSHR mRNA levels warrants further examination. The objectives of the present study were to clone and determine the complete FSHR cDNA sequence of the hamster and to delineate the effects of endogenous and exogenous FSH on the steady-state levels of ovarian FSHR mRNA. Complete FSHR cDNA was derived from hamster ovarian total RNA by the strategy of 3'- and 5'-rapid amplification of cDNA ends. Ovaries were obtained before and after the endogenous gonadotropin surge or exogenous FSH administration, and the steady-state levels of FSHR mRNA were assessed by Northern blot hybridization. Cloned FSHR cDNA consists of a reading frame corresponding to exons 1-10 of the human FSHR gene and the 5'- and 3'-untranslated regions. The nucleic acid and amino acid sequences of the reading frame were at least 87% and 92% identical, respectively, to that of human, rat, and mouse FSHR. Furthermore, the amino acid sequence contained seven transmembrane domains characteristic of the FSHR. The steady-state levels of FSHR mRNA increased from estrus (Day 1) to reach a peak on proestrus (Day 4) noon; however, significant attenuation was noted following the gonadotropin surge, which was blocked by phenobarbital. Exogenous FSH also downregulated, both dose- and time-dependently, ovarian FSHR mRNA levels. These data indicate that the nucleic acid sequence of hamster FSHR has been identified and that FSH modulates FSHR mRNA levels in the hamster ovary.
虽然有报道称促性腺激素可下调雌性大鼠卵巢中促卵泡激素受体(FSHR)mRNA水平,但促性腺激素高峰,尤其是促卵泡激素(FSH)对仓鼠卵泡FSHR mRNA水平的影响仍有待进一步研究。本研究的目的是克隆并确定仓鼠的完整FSHR cDNA序列,并描述内源性和外源性FSH对卵巢FSHR mRNA稳态水平的影响。通过3'和5' cDNA末端快速扩增策略从仓鼠卵巢总RNA中获得完整的FSHR cDNA。在内源性促性腺激素高峰之前和之后或外源性FSH给药前后获取卵巢,并通过Northern印迹杂交评估FSHR mRNA的稳态水平。克隆的FSHR cDNA由对应于人FSHR基因外显子1-10的阅读框以及5'和3'非翻译区组成。阅读框的核酸和氨基酸序列与人、大鼠和小鼠FSHR的核酸和氨基酸序列分别至少有87%和92%的同一性。此外,氨基酸序列包含FSHR特有的七个跨膜结构域。FSHR mRNA的稳态水平从发情期(第1天)开始升高,在发情前期(第4天)中午达到峰值;然而,在促性腺激素高峰后观察到明显衰减,苯巴比妥可阻断这种衰减。外源性FSH也剂量和时间依赖性地下调卵巢FSHR mRNA水平。这些数据表明已鉴定出仓鼠FSHR的核酸序列,并且FSH可调节仓鼠卵巢中FSHR mRNA水平。