Microarray analysis of the primate luteal transcriptome during chorionic gonadotrophin administration simulating early pregnancy
Microarray analysis of the primate luteal transcriptome during chorionic gonadotrophin administration simulating early pregnancy
复制标题
DOI：
10.1093/molehr/gar073
复制
发表时间：
2012-04-01
影响因子：
4
通讯作者：
Stouffer, R. L.
中科院分区：
文献类型：
--
作者：
Bishop, C. V.;Satterwhite, S.;Stouffer, R. L.
为探讨绒毛膜促性腺激素（CG）在灵长类动物黄体（CL）中的调控基因表达，在模拟早孕（SEP）模型中处理成年雌性恒河猴。在体内暴露于重组hCG的特定时间间隔（1、3、6和9天）后，从个体CL中分离总RNA，并与Affyssin基因芯片恒河猴基因组阵列杂交。与第10天未给药对照相比，SEP期间1192个转录物的mRNA水平变化了0.02倍[单因素ANOVA，错误发现率（FDR）校正; P 0.05]。实时荧光定量PCR验证结果表明，17个基因中有15个基因的表达模式与芯片结果相匹配。通过蛋白质印迹分析定量鉴定为CG敏感性的三个基因，CYP 19 A1（芳香化酶）、PGRMC 1（孕激素结合蛋白）和星星（类固醇生成急性调节蛋白）的蛋白质水平。为了进一步分析CG暴露诱导的基因表达的总体变化，利用先前储存的来自月经周期黄体期的基因芯片数据，比较SEP（挽救的）和消退CL之间的黄体基因表达。在时间匹配的间隔之间分析表达模式和mRNA水平。在hCG暴露（SEP）CL和回归CL之间，7677种mRNA的转录本在表达模式上存在差异，差异为2.2倍（单因素ANOVA，FDR校正; P 0.05）。回归CL（月经时）最不同于所有其他CL。进行显著受影响的转录物的途径分析;受CG暴露影响最大的途径是类固醇生物合成。进一步比较CG拯救和自然月经周期中黄体溶解过程中黄体基因表达的全基因组变化，应确定促进灵长类动物生育力的其他关键调控途径。
To explore chorionic gonadotrophin (CG)-regulated gene expression in the primate corpus luteum (CL), adult female rhesus macaques were treated in a model of simulated early pregnancy (SEP). Total RNA was isolated from individual CL after specific intervals of exposure (1, 3, 6 and 9 days) to recombinant hCG in vivo and hybridized to Affymetrix GeneChip Rhesus Macaque Genome Arrays. The mRNA levels of 1192 transcripts changed epsilon 2-fold [one-way ANOVA, false discovery rate (FDR) correction; P 0.05] during SEP when compared with Day 10 untreated controls. Real-time PCR validation indicated that 15 of 17 genes matched in expression pattern between PCR and microarray. Protein levels of three genes identified as CG-sensitive, CYP19A1 (aromatase), PGRMC1 (progestin-binding protein) and STAR (steroidogenic acute regulatory protein) were quantified by western blot analysis. To further analyze global changes in gene expression induced by CG exposure, luteal gene expression was compared between SEP (rescued) and regressing CL, utilizing previously banked GeneChip data from the luteal phase of the menstrual cycle. Expression patterns and mRNA levels were analyzed between time-matched intervals. Transcripts for 7677 mRNAs differed in expression patterns epsilon 2-fold (one-way ANOVA, FDR correction; P 0.05) between the hCG-exposed (SEP) CL and regressing CL. Regressed CL (at menses) were most unlike all other CL. Pathway analysis of significantly affected transcripts was performed; the pathway most impacted by CG exposure was steroid biosynthesis. Further comparisons of the genome-wide changes in luteal gene expression during CG rescue and luteolysis in the natural menstrual cycle should identify additional key regulatory pathways promoting primate fertility.