Hamernik D L, Clay C M, Turzillo A, Van Kirk E A, Moss G E
Department of Veterinary and Biomedical Science, University of Nebraska, Lincoln, USA.
Biol Reprod. 1995 Jul;53(1):179-85. doi: 10.1095/biolreprod53.1.179.
Two experiments were conducted simultaneously to investigate regulation of amounts of mRNA for GnRH receptors during the periovulatory period in sheep. In the first experiment, amounts of mRNA for GnRH receptors were measured before and after preovulatory surge of LH following regression of the CL by prostaglandin F2 alpha(PGF2 alpha). So that the time of the preovulatory surge of LH could be accurately predicted, ewes received two injections of PGF2 alpha on Day 14 of the estrous cycle. Anterior pituitary glands were collected from 5 control ewes on Day 14 of the estrous cycle (0 h after PGF2 alpha) and at 48, 72, and 96 h after PGF2 alpha (5 ewes per group). The second experiment was conducted to investigate the effects of 17 beta-estradiol on amounts of mRNA for GnRH receptors. On Day 14 of the estrous cycle, 20 ewes were ovariectomized (OVX); 15 of these ewes received estradiol implants when they were OVX (OVXEI). Sixteen hours after OVX, anterior pituitary glands were collected from 5 OVX and 5 OVXEI ewes, and the remaining OVXEI ewes received an i.m. injection of estradiol (25 micrograms in corn oil; OVXEI + E) to induce a preovulatory-like surge of LH. Anterior pituitary glands were collected from OVXEI + E ewes 18 or 54 h after injection of estradiol (n = 5 per group). Half of each anterior pituitary gland was used to measure the number of GnRH receptors. Poly(A)+ RNA was isolated from the remaining half of each anterior pituitary gland, applied to slot blots, and hybridized with a radioactive cDNA probe encoding the ovine GnRH receptor.(ABSTRACT TRUNCATED AT 250 WORDS)
同时进行了两项实验,以研究绵羊排卵期前后促性腺激素释放激素(GnRH)受体mRNA量的调节。在第一个实验中,通过前列腺素F2α(PGF2α)使黄体退化后,在促黄体生成素(LH)排卵前激增前后测量GnRH受体的mRNA量。为了能准确预测LH排卵前激增的时间,在发情周期的第14天给母羊注射两次PGF2α。在发情周期的第14天(PGF2α注射后0小时)以及PGF2α注射后48、72和96小时,从5只对照母羊收集垂体前叶(每组5只母羊)。进行第二个实验以研究17β-雌二醇对GnRH受体mRNA量的影响。在发情周期的第14天,对20只母羊进行卵巢切除(OVX);其中15只母羊在卵巢切除时植入雌二醇(OVXEI)。卵巢切除16小时后,从5只OVX母羊和5只OVXEI母羊收集垂体前叶,其余OVXEI母羊接受肌肉注射雌二醇(25微克溶于玉米油;OVXEI + E)以诱导类似排卵前的LH激增。在注射雌二醇后18或54小时从OVXEI + E母羊收集垂体前叶(每组n = 5)。每个垂体前叶的一半用于测量GnRH受体的数量。从每个垂体前叶的另一半分离出聚腺苷酸加尾(Poly(A)+)RNA,点样于印迹膜上,并用编码绵羊GnRH受体的放射性cDNA探针进行杂交。(摘要截短于250词)