Guy M K, Juengel J L, Tandeski T R, Niswender G D
Animal Reproduction and Biotechnology Laboratory, Department of Physiology, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, 80523, Fort Collins, Colorado, USA.
Endocrine. 1995 Aug;3(8):585-9. doi: 10.1007/BF02953023.
A partial cDNA was used to measure steady-state concentrations of mRNA encoding the receptor for luteinizing hormone (LH) in ovine corpora lutea. In experiment 1, luteal tissue and purified preparations of small and large steroidogenic luteal cells (n=4 per day) were obtained on days 3 (tissue only), 6, 9, 12 and 15 of the estrous cycle (estrus=day 0). Steady-state concentrations (fmoles receptor mRNA/μg poly(A)(+) RNA) and total quantities of mRNA (fmoles/corpus luteum) encoding the receptor for LH in luteal tissue increased (P<0.05) from day 3 to days 9 and 12 of the cycle; values on days 6 and 15 were intermediate. Small luteal cells contained at least four-fold greater (P<0.001) concentrations of mRNA encoding the receptor for LH than large luteal cells on days 6, 9, 12 and 15 of the cycle. In experiment 2, ewes on days 11 or 12 of the cycle received an infusion of either 1 μmol prostaglandin F(2α) (PGF(2α)) or saline into the ovarian artery. Luteal tissue was collected 1 (n=6), 4 (n=5), 12 (n=5) or 24 (n=5) h following PGF(2α) infusion, and 0 (no infusion;n=3), 12 (n=3) or 24 (n=4) h following saline administration. Concentrations of progesterone in sera decreased (P<0.05) within 12 h and remained low, whereas luteal weight and concentrations of progesterone in luteal tissue did not decrease (P<0.05) until 24 h after PGF(2α) treatment. Steady-state concentrations of mRNA encoding the receptor for LH were reduced (P<0.05) within 4 h of PGF(2α) infusion, and continued to decrease at 12 and 24 h post treatment. Calculated amounts of mRNA encoding the receptor for LH per corpus luteum were reduced (P<0.05) at 12 h after the PGF(2α) treatment and were 10% (P<0.05) of the values in saline-treated ewes at 24 h post-treatment. The increase during the estrous cycle in steady-state concentrations of mRNA encoding the receptor for LH appears to occur prior to the previously observed increase in number of receptors for LH. Following PGF(2α)-induced luteal regression, concentrations of mRNA encoding LH receptor decreased prior to the previously reported decrease in LH binding. Thus, changes in the number of receptors for LH in ovine luteal tissue during luteal development and luteolysis appears to be preceded by corresponding changes in mRNA encoding this receptor.
利用部分互补DNA(cDNA)来测定绵羊黄体中编码促黄体生成素(LH)受体的信使核糖核酸(mRNA)的稳态浓度。在实验1中,于发情周期的第3天(仅取组织)、第6天、第9天、第12天和第15天(发情日为第0天)获取黄体组织以及纯化的小、大两种类固醇生成黄体细胞制剂(每天n = 4)。黄体组织中编码LH受体的mRNA的稳态浓度(飞摩尔受体mRNA/微克聚腺苷酸[poly(A)(+)]RNA)和mRNA总量(飞摩尔/黄体)在发情周期从第3天到第9天和第12天增加(P<0.05);第6天和第15天的值介于两者之间。在发情周期的第6天、第9天、第12天和第15天,小黄体细胞中编码LH受体的mRNA浓度比大黄体细胞至少高4倍(P<0.001)。在实验2中,处于发情周期第11天或第12天的母羊经卵巢动脉输注1微摩尔前列腺素F2α(PGF2α)或生理盐水。在输注PGF2α后1小时(n = 6)、4小时(n = 5)、12小时(n = 5)或24小时(n = 5)采集黄体组织,在输注生理盐水后0小时(未输注;n = 3)、12小时(n = 3)或24小时(n = 4)采集。血清中孕酮浓度在12小时内下降(P<0.05)并维持在低水平,而黄体重量和黄体组织中孕酮浓度直到PGF2α处理后24小时才下降(P<0.05)。编码LH受体的mRNA的稳态浓度在PGF2α输注后4小时内降低(P<0.05),并在处理后12小时和24小时持续下降。计算得出的每个黄体中编码LH受体的mRNA量在PGF2α处理后12小时减少(P<0.05),在处理后24小时为生理盐水处理母羊值的10%(P<0.05)。发情周期中编码LH受体的mRNA稳态浓度的增加似乎发生在先前观察到的LH受体数量增加之前。在PGF2α诱导黄体退化后,编码LH受体的mRNA浓度在先前报道的LH结合减少之前下降。因此,在黄体发育和黄体溶解过程中,绵羊黄体组织中LH受体数量的变化似乎之前有编码该受体的mRNA的相应变化。