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未成熟和成熟公猪睾丸中雌激素受体α和β以及芳香化酶的表达

Expression of oestrogen receptors alpha and beta and of aromatase in the testis of immature and mature boars.

作者信息

Mutembei H M, Pesch S, Schuler G, Hoffmann B

机构信息

Clinic for Obstetrics, Gynaecology and Andrology of Large and Small Animals, Justus-Liebig-University, Giessen, Germany.

出版信息

Reprod Domest Anim. 2005 Jun;40(3):228-36. doi: 10.1111/j.1439-0531.2005.00586.x.

Abstract

The boar testis secretes high amounts of oestrogens. In order to test for a likely local significance, we investigated the expression of oestrogen receptors (ER) in immature and mature boar testes using immunohistochemistry (IHC), in vitro and in situ reverse transcription-polymerase chain reaction (RT-PCR). Samples were from 25 boars castrated at ages of 50, 100, 150, 200 and 250 days. Mouse monoclonal primary antibodies against porcine ERalpha (clone HT227), human ERbeta1 (clone PPG5/10) and human P450 aromatase (clone SM1671P) were used. Expression of the mRNA was tested utilizing primers specific for the respective porcine mRNA sequences. ER immunoreactivity was exclusively localized to the nuclei. In immature boars, 90.6 +/- 1.2% of prespermatogonia and 71.0 +/- 2.6% of the Leydig cells showed a strong staining for ERalpha; 95.5 +/- 3.5% of the prespermatogonia but none of the Leydig and Sertoli cells were ERbeta-positive. In mature boars a strong staining for the ERbeta was observed in virtually all Sertoli, Leydig and germ cells, except for the elongating/ed spermatids, which were clearly negative; for the ERalpha, strong immunoreaction signals were restricted to spermatogonia and primary spermatocytes with 93.6 +/- 2.7% of these cells being positive; distinctly less intensive signals were observed in 51.4 +/- 0.27% of the secondary spermatocytes, round spermatids and Leydig cells. In vitro RT-PCR was positive for both receptors and results of in situ RT-PCR matched those obtained by IHC. P450 aromatase immunoreaction was restricted to the cytoplasm of Leydig cells. These findings suggest that testicular ER may be important factors contributing to onset and maintenance of spermatogenesis in the boar.

摘要

公猪睾丸分泌大量雌激素。为了检测其可能的局部意义,我们采用免疫组织化学(IHC)、体外和原位逆转录-聚合酶链反应(RT-PCR),研究了未成熟和成熟公猪睾丸中雌激素受体(ER)的表达。样本取自25头分别在50、100、150、200和250日龄时被阉割的公猪。使用了针对猪ERα(克隆HT227)、人ERβ1(克隆PPG5/10)和人P450芳香化酶(克隆SM1671P)的小鼠单克隆一抗。利用针对各自猪mRNA序列的特异性引物检测mRNA的表达。ER免疫反应仅定位于细胞核。在未成熟公猪中,90.6±1.2%的精原细胞前体细胞和71.0±2.6%的睾丸间质细胞对ERα呈强染色;95.5±3.5%的精原细胞前体细胞为ERβ阳性,但睾丸间质细胞和支持细胞均无阳性。在成熟公猪中,除了正在伸长/成熟的精子细胞明显为阴性外,几乎所有支持细胞、睾丸间质细胞和生殖细胞中均观察到ERβ的强染色;对于ERα,强免疫反应信号仅限于精原细胞和初级精母细胞,其中93.6±2.7%的细胞呈阳性;在51.4±0.27%的次级精母细胞、圆形精子细胞和睾丸间质细胞中观察到明显较弱的信号。体外RT-PCR检测两种受体均呈阳性,原位RT-PCR结果与IHC结果相符。P450芳香化酶免疫反应仅限于睾丸间质细胞的细胞质。这些发现表明,睾丸ER可能是对公猪精子发生的起始和维持起重要作用的因素。

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