Masri B A, Russell L D, Vogl A W
Anat Rec. 1987 May;218(1):20-6. doi: 10.1002/ar.1092180105.
In this study, we describe the distribution of actin filaments in and around spermatids that are mechanically dissociated, in the presence and absence of exogenous trypsin, from the seminiferous epithelium of the rat. NBD-phallacidin and subfragment 1 of the myosin molecule (S-1) are used as probes for filamentous actin at the light and electron microscopic levels, respectively. The fluorescence associated with spermatids mechanically dissociated in the absence of trypsin is due to actin both in the spermatogenic cells themselves and in attached Sertoli cell ectoplasmic specializations. Fluorescence generated by labelled actin in ectoplasmic specializations occurs in linear tracts that follow the outer contour of spermatid heads. The residual fluorescence seen when trypsin is used to detach Sertoli cell fragments is diffuse and due to f-actin in the subacrosomal space. Electron microscopic data agree with the fluorescence results. This study conclusively demonstrates that Sertoli cell ectoplasmic specializations remain attached to spermatids mechanically dissociated from the seminiferous epithelium. This observation may be useful when attempting to isolate ectoplasmic specializations for biochemical analyses.
在本研究中，我们描述了在有或没有外源性胰蛋白酶存在的情况下，从大鼠生精上皮机械解离的精子细胞及其周围肌动蛋白丝的分布。在光镜和电镜水平上，分别使用NBD-鬼笔环肽和肌球蛋白分子的亚片段1（S-1）作为丝状肌动蛋白的探针。在没有胰蛋白酶的情况下机械解离的精子细胞相关荧光，是由于生精细胞自身以及附着的支持细胞外质特化结构中的肌动蛋白所致。外质特化结构中标记肌动蛋白产生的荧光出现在沿着精子细胞头部外轮廓的线性束中。当使用胰蛋白酶分离支持细胞片段时看到的残留荧光是弥散的，并且是由于顶体下空间中的丝状肌动蛋白。电镜数据与荧光结果一致。本研究确凿地证明，支持细胞外质特化结构在从生精上皮机械解离的精子细胞上仍然附着。当试图分离外质特化结构进行生化分析时，这一观察结果可能会有用。
