Igarashi-Migitaka Junko, Takeshita Akira, Koibuchi Noriyuki, Yamada Shozo, Ohtani-Kaneko Ritsuko, Hirata Kazuaki
Department of Anatomy and Cell Biology, St Marianna University School of Medicine, Kawasaki, Kanagawa 216-8511, Japan.
Eur J Endocrinol. 2005 Oct;153(4):595-604. doi: 10.1530/eje.1.01990.
Androgens are critical for the development and maintenance of male sexual characteristics. Their action is mediated through the androgen receptor (AR). Ligand-bound AR interacts with coactivator proteins that mediate transcriptional activation. Such coactivators include three members of the 160 kDa proteins (p160s): SRC-1, TIF2/GRIP1, and p/CIP/RAC3/ACTR/AIB1/TRAM-1. The aim of this study was to investigate the expression of the three p160 coactivators and their association with AR in testis and epididymis.
We determined the localization of these three p160 coactivators in immature and mature rat testis, and epididymis by immunohistochemistry using the specific monoclonal antibodies. We also performed double immunofluorescence staining to examine whether p160s are colocalized with AR in these tissues.
In seminiferous tubules of mature rat testis, SRC-1 and TRAM-1 immunoreactivity was found predominantly in spermatogonia and spermatocytes. In contrast, TIF2 was expressed predominantly in Sertoli cells. AR was coexpressed with TIF2 in this cell type. In immature rat testis, however, all three coactivators were expressed in both germ cells and Sertoli cells. In the epididymis, SRC-1 and TIF2 immunoreactivities were localized in nuclei of epithelial cells. However, TRAM-1 immunostaining was observed in the luminal portion of the cytoplasm with greater intensity than in the nucleus, especially in the caput epididymidis.
The cell-type-specific expression of p160 coactivators suggests specific roles in male reproductive organs. Further, the strong cytoplasmic localization of TRAM-1 protein in epithelial cells of epididymis suggests that TRAM-1 may have additional role(s) in transcriptional regulation.
雄激素对于男性性征的发育和维持至关重要。其作用通过雄激素受体(AR)介导。配体结合的AR与介导转录激活的共激活蛋白相互作用。此类共激活蛋白包括160 kDa蛋白(p160s)的三个成员:SRC-1、TIF2/GRIP1和p/CIP/RAC3/ACTR/AIB1/TRAM-1。本研究的目的是调查这三种p160共激活蛋白在睾丸和附睾中的表达及其与AR的关联。
我们使用特异性单克隆抗体通过免疫组织化学确定这三种p160共激活蛋白在未成熟和成熟大鼠睾丸及附睾中的定位。我们还进行了双重免疫荧光染色,以检查p160s是否与这些组织中的AR共定位。
在成熟大鼠睾丸的生精小管中,SRC-1和TRAM-1免疫反应性主要在精原细胞和精母细胞中发现。相比之下,TIF2主要在支持细胞中表达。AR与TIF2在这种细胞类型中共表达。然而,在未成熟大鼠睾丸中,所有三种共激活蛋白在生殖细胞和支持细胞中均有表达。在附睾中,SRC-1和TIF2免疫反应性定位于上皮细胞核中。然而,TRAM-1免疫染色在细胞质的管腔部分观察到,其强度大于细胞核,尤其是在附睾头。
p160共激活蛋白的细胞类型特异性表达表明其在男性生殖器官中具有特定作用。此外,TRAM-1蛋白在附睾上皮细胞中的强细胞质定位表明TRAM-1可能在转录调控中具有额外作用。