Fiedler E P, Plouffe L, Hales D B, Hales K H, Khan I
Reproductive Endocrinology, Infertility and Genetics Section, Department of Obstetrics and Gynecology, The Medical College of Georgia, Augusta, Georgia 30912, USA.
Biol Reprod. 1999 Sep;61(3):643-50. doi: 10.1095/biolreprod61.3.643.
With interest in steroidogenic acute regulatory protein (StAR) involvement in the luteolytic process, we studied changes in serum progesterone levels and the concomitant expression of StAR mRNA and protein (37-, 32-, and 30-kDa forms) in postovulatory Day 7 corpora lutea (CL) isolated from rats 1 h after injection with prostaglandin F(2alpha) (PGF(2alpha), n = 6) or saline (n = 6). Serum progesterone levels were determined by RIA, StAR and beta-actin mRNA expression by Northern analysis, and StAR and beta-actin protein expression by Western analysis. Adrenal, brain, and spleen from control animals were used as positive and negative controls for StAR expression. Scanning optical densitometry measurements were standardized by dividing the signal strength from each StAR autoradiogram lane by that from the corresponding beta-actin autoradiogram lane. ANOVA was used for significance testing, with alpha set at 0.05. The 37-, 32-, and 30-kDa forms of StAR protein were expressed in all adrenal samples, whereas only the 37- and 30-kDa forms were found in CL. Serum progesterone levels and expression of the 30-kDa and 37-kDa forms of the StAR protein in CL were all found to be significantly lower in the PGF(2alpha)-treated than the saline-treated group. StAR mRNA expression was not significantly different in the saline- and PGF(2alpha)-treated rats. The rapid decline in StAR protein expression that accompanies PGF(2alpha) induced luteolysis, therefore, does not result from significant decline in mRNA expression.
鉴于对类固醇生成急性调节蛋白(StAR)参与黄体溶解过程的研究兴趣,我们研究了从注射前列腺素F(2α)(PGF(2α),n = 6)或生理盐水(n = 6)1小时后的大鼠分离出的排卵后第7天黄体(CL)中血清孕酮水平的变化以及StAR mRNA和蛋白(37 kDa、32 kDa和30 kDa形式)的伴随表达。通过放射免疫分析法测定血清孕酮水平,通过Northern分析测定StAR和β-肌动蛋白mRNA表达,通过Western分析测定StAR和β-肌动蛋白蛋白表达。将对照动物的肾上腺、脑和脾脏用作StAR表达的阳性和阴性对照。通过将每个StAR放射自显影片条带的信号强度除以相应的β-肌动蛋白放射自显影片条带的信号强度来对扫描光密度测量进行标准化。使用方差分析进行显著性检验,α设定为0.05。StAR蛋白的37 kDa、32 kDa和30 kDa形式在所有肾上腺样品中均有表达,而在CL中仅发现37 kDa和30 kDa形式。发现PGF(2α)处理组CL中的血清孕酮水平以及StAR蛋白30 kDa和37 kDa形式的表达均显著低于生理盐水处理组。在生理盐水处理和PGF(2α)处理的大鼠中,StAR mRNA表达没有显著差异。因此,PGF(2α)诱导黄体溶解时伴随的StAR蛋白表达的快速下降并非由mRNA表达的显著下降所致。