Manikkam M, Bao B, Rosenfeld C S, Yuan X, Salfen B E, Calder M D, Youngquist R S, Keisler D H, Lubahn D B, Garverick H A
Department of Animal Sciences, University of Missouri, East Campus Dr., 163 Animal Science Research Center, Columbia, MO 65211-5300, USA.
Anim Reprod Sci. 2001 Sep 15;67(3-4):159-69. doi: 10.1016/s0378-4320(01)00121-x.
In a previous study, the ERbeta cDNA protein-coding region was utilised to clone bovine ERbeta. The objectives in this study were to examine (1) ERbeta mRNA expression in ovarian follicles throughout the bovine first follicular wave, and (2) effect of LH infusion into cows on bERbeta mRNA expression during the second follicular wave. In experiment 1, heifers (4-5 per time point) were ovariectomized at 12, 24, 36, 48, 60, 72, 84, 96, 144, or 216 h after emergence of the first follicular wave after oestrus. In experiment 2, saline or LH was pulsed hourly (computer-controlled syringe pump) into cows (n = 31; 5-6 per treatment) at wave emergence for 2 or 4 days: wave 1-saline (W1S), wave 2-saline (W2S), or wave 2-LH (25 microg/h; W2LH). Ovaries were removed on day 2 or day 4 after wave emergence. Follicles, 2-19mm in size, were dissected, frozen, and stored at -80 degrees C for in situ hybridisation with two bERbeta cRNA probes. Expression of bERbeta mRNA was localised in granulosa cells of healthy follicles. In experiment 1, bERbeta mRNA expression did not change with time points of the wave showing no association of bERbeta mRNA expression with follicular selection and dominance. However, bERbeta mRNA expression decreased with increase in size of all follicles. Expression of bERbeta mRNA was greater in very small follicles (2-4 mm) than in large (> or = 9 mm) follicles. In experiment 2, expression of bERbeta mRNA in follicles did not differ either between W1S and W2S or between W2S and W2LH. In summary, bERbeta mRNA expression decreased with increasing follicular size. However, neither stage of the wave (selection or dominance), nor pulsatile infusion of LH influenced bERbeta mRNA expression.
在之前的一项研究中,利用雌激素受体β(ERβ)的cDNA蛋白编码区克隆了牛的ERβ。本研究的目的是检测:(1)在整个牛的第一个卵泡波期间,卵巢卵泡中ERβ mRNA的表达情况;(2)在第二个卵泡波期间,给母牛注射促黄体素(LH)对牛ERβ mRNA表达的影响。在实验1中,在发情后第一个卵泡波出现后的12、24、36、48、60、72、84、96、144或216小时,对小母牛(每个时间点4 - 5头)进行卵巢切除。在实验2中,在卵泡波出现时,用生理盐水或LH(通过计算机控制的注射泵)每小时脉冲注射一次,对母牛(n = 31;每种处理5 - 6头)进行2天或4天的处理:第1波 - 生理盐水(W1S),第2波 - 生理盐水(W2S),或第2波 - LH(25微克/小时;W2LH)。在卵泡波出现后的第2天或第4天切除卵巢。将大小为2 - 19毫米的卵泡进行解剖、冷冻,并储存在 - 80℃,用于与两种牛ERβ cRNA探针进行原位杂交。bERβ mRNA的表达定位于健康卵泡的颗粒细胞中。在实验1中,bERβ mRNA的表达在卵泡波的各个时间点没有变化,表明bERβ mRNA的表达与卵泡的选择和优势化无关。然而,bERβ mRNA的表达随着所有卵泡大小的增加而降低。在非常小的卵泡(2 - 4毫米)中,bERβ mRNA的表达高于大卵泡(≥9毫米)。在实验2中,W1S和W2S之间或W2S和W2LH之间,卵泡中bERβ mRNA的表达没有差异。总之,bERβ mRNA的表达随着卵泡大小的增加而降低。然而,卵泡波的阶段(选择或优势化)以及LH的脉冲注射均未影响bERβ mRNA的表达。