Pelletier R, Trifaro J M, Carbajal M E, Okawara Y, Vitale M L
Department of Pathology and Cell Biology, Faculty of Medicine, Université de Montréal, P. Québec, Canada H3T 1J4.
Biol Reprod. 1999 May;60(5):1128-36. doi: 10.1095/biolreprod60.5.1128.
We assessed the levels and localization of the actin filament-severing protein scinderin, in fetal and adult bovine testes, and in spermatozoa during and following the epididymal transit. We performed immunoblots on seminiferous tubules and interstitial cells isolated by enzymatic digestion, and on bovine chromaffin cells, spermatozoa, aorta, and vena cava. Immunoperoxidase labeling was done on Bouin's perfusion-fixed testes and epididymis tissue sections, and on spermatozoa. In addition, immunofluorescence labeling was done on spermatozoa. Immunoblots showed one 80-kDa band in chromaffin cells, fetal and adult tubules, interstitial cells, spermatozoa, aorta, and vena cava. Scinderin levels were higher in fetal than in adult seminiferous tubules but showed no difference between fetal and adult interstitial cells. Scinderin levels were higher in epididymal than in ejaculated spermatozoa. Scinderin was detected in a region corresponding with the subacrosomal space in the round spermatids and with the acrosome in the elongated spermatids. In epididymal spermatozoa, scinderin was localized to the anterior acrosome and the equatorial segment, but in ejaculated spermatozoa, the protein appeared in the acrosome and the post-equatorial segment of the head. In Sertoli cells, scinderin was detected near the cell surface and within the cytoplasm, where it accumulated near the base in a stage-specific manner. In the epididymis, scinderin was localized next to the surface of the cells; in the tail, it collected near the base of the principal cells. In Sertoli cells and epididymal cells, scinderin may contribute to the regulation of tight junctional permeability and to the release of the elongated spermatids by controlling the state of perijunctional actin. In germ cells, scinderin may assist in the shaping of the developing acrosome and influence the fertility of the spermatozoa.
我们评估了肌动蛋白丝切断蛋白scinderin在胎儿和成年牛睾丸以及附睾转运期间和之后的精子中的水平和定位。我们对通过酶消化分离的生精小管和间质细胞、牛嗜铬细胞、精子、主动脉和腔静脉进行了免疫印迹分析。对经Bouin氏液灌注固定的睾丸和附睾组织切片以及精子进行了免疫过氧化物酶标记。此外,对精子进行了免疫荧光标记。免疫印迹显示,嗜铬细胞、胎儿和成年小管、间质细胞、精子、主动脉和腔静脉中有一条80 kDa的条带。胎儿生精小管中的scinderin水平高于成年生精小管,但胎儿和成年间质细胞之间没有差异。附睾精子中的scinderin水平高于射出精子。在圆形精子细胞中,scinderin在与顶体下间隙相对应的区域被检测到,在长形精子细胞中则与顶体相对应。在附睾精子中,scinderin定位于顶体前部和赤道段,但在射出精子中,该蛋白出现在头部的顶体和赤道后段。在支持细胞中,scinderin在细胞表面附近和细胞质内被检测到,在细胞质内它以阶段特异性方式在基部附近积累。在附睾中,scinderin定位于细胞表面旁边;在尾部,它聚集在主细胞基部附近。在支持细胞和附睾细胞中,scinderin可能通过控制连接周肌动蛋白的状态,有助于调节紧密连接的通透性,并有助于长形精子细胞的释放。在生殖细胞中,scinderin可能有助于发育中的顶体的形成,并影响精子的受精能力。