Juengel J L, Meberg B M, Turzillo A M, Nett T M, Niswender G D
Animal Reproduction and Biotechnology Laboratory, Colorado State University, Fort Collins 80523-1683, USA.
Endocrinology. 1995 Dec;136(12):5423-9. doi: 10.1210/endo.136.12.7588291.
Steroidogenic acute regulatory protein (StAR), proposed to be involved in the transport of cholesterol to the inner mitochondrial membrane, has recently been cloned from MA-10 cells. Using reverse transcription-polymerase chain reaction, we generated a complementary DNA encoding 404 base pairs of StAR from ovine luteal tissue to perform studies regarding regulation of the messenger RNA (mRNA) encoding this protein. In Exp 1, ewes were hypophysectomized (HPX) on day 5 of the estrous cycle and administered saline or physiological regimens of LH and/or GH until collection of luteal tissue on day 12 of the estrous cycle (n = 4/group). Luteal concentrations [mean +/- SEM; femtomoles per microgram poly(A)+ RNA] of mRNA encoding StAR were lower (P < 0.05) in the HPX plus saline-treated ewes (26.4 +/- 7.3) than in day 12 pituitary-intact ewes (n = 4; 77.7 +/- 9.3). Replacement of LH (59.1 +/- 13.1), GH (59.1 +/- 12.8), or LH and GH (69.9 +/- 4.5) in HPX ewes increased (P < 0.05) concentrations of mRNA encoding StAR to values not different from those in day 12 controls. In Exp 2, ewes on day 11 or 12 of the estrous cycle were injected with prostaglandin F2 alpha (PGF2 alpha) to induce luteal regression. Corpora lutea were collected 4, 12, or 24 h after injection (n = 4-5/time point) and from untreated control ewes (n = 4) or 24 h after injection of saline (n = 4). Treatment with PGF2 alpha decreased (P < 0.05) concentrations of progesterone in serum 4, 12, and 24 h after injection. Concentrations of StAR mRNA were decreased (P < 0.01) to 47%, 19%, and 8% of control values 4, 12, and 24 h after PGF2 alpha injection, respectively. In Exp 3, ewes received ovarian arterial infusions of saline, PGF2 alpha, or phorbol 12-myristate 13-acetate (PMA), and luteal tissue was collected 0 (no infusion), 4, 12, or 24 h later (n = 3-4/group). Treatment with PGF2 alpha or PMA decreased (P < 0.05) concentrations of progesterone in serum 4, 12, and 24 h postinjection. Steady state concentrations of mRNA encoding StAR (P < 0.05) were 36% and 25% of the control value 12 and 24 h after PGF2 alpha injection. Injection of PMA decreased (P < 0.05) concentrations of StAR mRNA to 75% and 50% of control values at 4 and 12 h, but concentrations of mRNA encoding StAR were not different from control values at 24 h.(ABSTRACT TRUNCATED AT 400 WORDS)
类固醇生成急性调节蛋白(StAR)被认为参与胆固醇向线粒体内膜的转运,最近已从MA - 10细胞中克隆出来。我们利用逆转录 - 聚合酶链反应,从绵羊黄体组织中生成了编码404个碱基对的StAR互补DNA,以开展关于编码该蛋白的信使核糖核酸(mRNA)调控的研究。在实验1中,母羊在发情周期的第5天接受垂体切除(HPX),并给予生理盐水或促黄体生成素(LH)和/或生长激素(GH)的生理剂量方案,直至在发情周期的第12天采集黄体组织(每组n = 4)。与发情周期第12天垂体完整的母羊(n = 4;77.7±9.3)相比,HPX加生理盐水处理的母羊中编码StAR的mRNA的黄体浓度[平均值±标准误;飞摩尔每微克聚腺苷酸(A)+ RNA]较低(P < 0.05)(26.4±7.3)。在HPX母羊中补充LH(59.1±13.1)、GH(59.1±12.8)或LH和GH(69.9±4.5)可使编码StAR的mRNA浓度增加(P < 0.05),达到与发情周期第12天对照组无差异的值。在实验2中,对发情周期第11天或第12天的母羊注射前列腺素F2α(PGF2α)以诱导黄体退化。在注射后4、12或24小时采集黄体(每个时间点n = 4 - 5),并从未经处理的对照母羊(n = 4)或注射生理盐水后24小时的母羊(n = 4)采集。PGF2α处理使注射后4、12和24小时血清中的孕酮浓度降低(P < 0.05)。PGF2α注射后4、12和24小时,StAR mRNA浓度分别降至对照值的47%、19%和8%(P < 0.01)。在实验3中,母羊接受卵巢动脉输注生理盐水、PGF2α或佛波酯12 - 肉豆蔻酸13 - 乙酸酯(PMA),并在0(未输注)[注:原文此处有误,应为“0 h(未输注)”]、4、12或24小时后采集黄体组织(每组n = 3 - 4)。PGF2α或PMA处理使注射后4、12和24小时血清中的孕酮浓度降低(P < 0.05)。PGF2α注射后12和24小时,编码StAR的mRNA的稳态浓度(P < 0.05)分别为对照值的36%和25%。注射PMA使4和12小时时StAR mRNA浓度降至对照值的75%和50%(P < 0.05),但在24小时时编码StAR的mRNA浓度与对照值无差异。(摘要截短于400字)