García E M, Vázquez J M, Parrilla I, Ortega M D, Calvete J J, Sanz L, Martínez E A, Roca J, Rodríguez-Martínez H
Department of Medicine and Animal Surgery, Faculty of Veterinary Medicine, University of Murcia, Murcia, Spain.
Int J Androl. 2008 Aug;31(4):408-17. doi: 10.1111/j.1365-2605.2007.00784.x.
The epithelial localization and expression of the spermadhesin PSP-I and PSP-II subunits were determined in the testis, ductus epididymes (caput, corpus and cauda), seminal vesicles and bulbourethral glands of mature boars, using immunohistochemical, western blotting and RT-PCR methods. Immunohistochemistry showed positive labelling for PSP-I and PSP-II antibodies in the epithelium of seminal vesicles in all males tested. Positive immunolabelling, but with variable intensity, was also present in the epididymal epithelium (caput, corpus and cauda), although varying largely among segments and boars. Immunoreactivity was nearly or completely absent in the seminiferous epithelium and the bulbourethral gland, although SDS-PAGE and western blotting revealed the presence of PSP-I and PSP-II immunoreactive bands in all the tissue extracts, including the testis and the bulbourethral gland. mRNA amplification by RT-PCR using primers specific for PSP-I and PSP-II showed a trend similar to that observed for western blotting, i.e. intensity variation between tissues (even between segments of the same epididymis) and among boars. Our results indicate that the seminal vesicles are the main source of PSP-I and PSP-II spermadhesins, although epididymal segments, testis and the bulbourethral gland also participate in the expression of both proteins.
采用免疫组织化学、蛋白质免疫印迹和逆转录聚合酶链反应(RT-PCR)方法,确定了精子黏附素PSP-I和PSP-II亚基在成年公猪睾丸、附睾管(头、体和尾)、精囊和尿道球腺中的上皮定位及表达情况。免疫组织化学显示,在所检测的所有雄性动物的精囊上皮中,PSP-I和PSP-II抗体呈阳性标记。附睾上皮(头、体和尾)中也存在阳性免疫标记,但强度各异,且在不同节段和公猪之间差异很大。生精上皮和尿道球腺中几乎没有或完全没有免疫反应性,尽管十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质免疫印迹显示,在包括睾丸和尿道球腺在内的所有组织提取物中均存在PSP-I和PSP-II免疫反应条带。使用针对PSP-I和PSP-II的特异性引物进行RT-PCR扩增mRNA,结果显示出与蛋白质免疫印迹相似的趋势,即不同组织(甚至同一附睾的不同节段)以及不同公猪之间强度存在差异。我们的结果表明,精囊是PSP-I和PSP-II精子黏附素的主要来源,尽管附睾节段、睾丸和尿道球腺也参与这两种蛋白质的表达。