Expression and molecular characterization of estrogen receptor alpha messenger RNA in male reproductive organs of adult goats.
Expression and molecular characterization of estrogen receptor alpha messenger RNA in male reproductive organs of adult goats.
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成年山羊雄性生殖器官中雌激素受体α信使RNA的表达和分子特征。
DOI：
10.1095/biolreprod64.5.1432
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发表时间：
2001
影响因子：
3.6
通讯作者：
Goyal,HO
中科院分区：
文献类型：
--
作者：
Mansour,MM;Machen,MR;Tarleton,BJ;Wiley,AA;Wower,J;Bartol,FF;Goyal,HO
雄性雌激素受体α（ERα）敲除小鼠不育的事实表明该受体在雄性生殖中的作用。本研究采用RNA酶保护试验（RPA）和原位杂交（ISH）技术，从转录水平检测雌激素受体α（ERα）在雄性山羊生殖组织中的表达，并采用逆转录-聚合酶链反应（RT-PCR）技术克隆山羊ERα的部分cDNA。对于RPA和ISH程序，使用了从绵羊oER 8 cDNA模板体外生成的放射性标记的反义cRNA探针。对从成年山羊获得的个体样品进行评价。标记的cRNA正义探针在ISH中用作阴性对照。通过RT-PCR从传出小管（ED）、附睾（EP）和睾丸中产生530个碱基对的扩增子，从艾德和EP中克隆，并测序。山羊雌激素受体α（cERα）cDNA序列与其它物种的同源性为81%~ 96%。RT-PCR和RPA均检测到ERα mRNA在所有组织中的表达，但在ED中表达最强。与艾德相比，ERα在EP中的表达低6倍，在睾丸中低66倍。同样，ISH在艾德上皮中观察到强ERα表达，而在EP或睾丸中检测到很少或没有信号。因此，在雄性生殖道和睾丸的不同节段中，ED上皮中ERα mRNA表达水平最高。
The fact that male estrogen receptor alpha (ERα) knockout mice are infertile indicates a role for this receptor in male reproduction. Here, objectives were to evaluate ERα expression in male goat reproductive tissues at the transcriptional level using RNase protection assay (RPA) and in situ hybridization (ISH), and to clone a partial cDNA for caprine ERα using reverse transcription-polymerase chain reaction (RT-PCR). For RPA and ISH procedures, a radiolabeled antisense cRNA probe, generated in vitro from the ovine oER8 cDNA template, was employed. Evaluations were made on individual samples obtained from adult goats. Labeled cRNA sense probe was used as a negative control in ISH. A 530-base pair amplicon was generated by RT-PCR from efferent ductules (EDs), epididymis (EP), and testis, cloned from the ED and EP, and sequenced. The caprine ERα (cERα) cDNA displayed 81%–96% sequence identity with that of other species. A signal indicative of ERα mRNA was identified by both RT-PCR and RPA in all tissues, but was strongest in the ED. Compared with ED, ERα signal was sixfold lower in the EP, and 66-fold lower in the testis. Similarly, strong ERα expression was observed in ED epithelium, whereas little or no signal was detected in EP or testis by ISH. Thus, among different segments of the male reproductive tract and testis, the highest level of ERα mRNA expression was found in epithelium of the ED.