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大鼠支持细胞和生精细胞表达一种相似的基因,其产物在抗原性上与一种外致密纤维相关蛋白有关。

Rat Sertoli and spermatogenic cells express a similar gene, and its product is antigenically related to an outer dense fiber-associated protein.

作者信息

Smith F F, Mertz J R, Krebs I, Tres L L, Chae C B, Zakeri Z, Engelhardt J, Hoover D, Tenniswood M, Kierszenbaum A L

机构信息

Department of Cell Biology and Anatomical Sciences, City University of New York (CUNY) Medical School, NY 10031.

出版信息

Mol Reprod Dev. 1992 Dec;33(4):363-72. doi: 10.1002/mrd.1080330402.

Abstract

We have previously reported that a heterodimeric protein secreted by rat Sertoli cells is antigenically related to a protein associated with outer dense fibers of the sperm tail. Therefore, we have explored the possibility that Sertoli and spermatogenic cells express a similar gene encoding a homologous protein. A Sertoli cell heterodimeric protein cDNA probe recognizes specific mRNA in pachytene and round spermatids fractionated by centrifugal elutriation; however, this specific mRNA was less prominent than in cultured Sertoli cells. In agreement with these observations, in situ hybridization experiments show that Sertoli cells are predominantly engaged in active heterodimeric protein mRNA synthesis, while meiotic prophase spermatocytes and spermatids also show significant but less abundant specific mRNA. Immunoblotting experiments demonstrate that, while Sertoli cells synthesize a heterodimeric protein consisting of two disulfide-linked components with molecular masses of 45 and 35 kD, both primary spermatocytes and round spermatids synthesize single 30 kD monomers not associated by disulfide linkage but recognized by antisera to Sertoli cell heterodimeric protein. Immunoblotting and immunogold electron microscopic studies show that antisera to Sertoli cell heterodimeric protein recognize a protein associated with outer dense fibers. This immunoreactivity was abolished by a 5-min pronase treatment, without affecting the integrity of outer dense fibers. Results of this study and previous studies demonstrate that both Sertoli and spermatogenic cells express a similar gene and that an antigenically related product encoded by this gene becomes associated with outer dense fibers during their assembly at spermiogenesis.

摘要

我们之前报道过,大鼠支持细胞分泌的一种异二聚体蛋白在抗原性上与精子尾部外致密纤维相关蛋白有关。因此,我们探讨了支持细胞和生精细胞是否表达编码同源蛋白的相似基因这一可能性。一种支持细胞异二聚体蛋白cDNA探针可识别通过离心淘析分离的粗线期和圆形精子细胞中的特异性mRNA;然而,这种特异性mRNA在这些细胞中不如在培养的支持细胞中明显。与这些观察结果一致,原位杂交实验表明,支持细胞主要参与活性异二聚体蛋白mRNA的合成,而减数分裂前期的精母细胞和精子细胞也显示有显著但含量较少的特异性mRNA。免疫印迹实验表明,支持细胞合成一种由两个通过二硫键连接的分子量分别为45kD和35kD的成分组成的异二聚体蛋白,而初级精母细胞和圆形精子细胞都合成单一的30kD单体,这些单体不是通过二硫键连接的,但能被针对支持细胞异二聚体蛋白的抗血清识别。免疫印迹和免疫金电子显微镜研究表明,针对支持细胞异二聚体蛋白的抗血清可识别一种与外致密纤维相关的蛋白。这种免疫反应性在经5分钟的链霉蛋白酶处理后消失,而不影响外致密纤维的完整性。本研究及之前研究的结果表明,支持细胞和生精细胞都表达相似的基因,并且在精子发生过程中,由该基因编码的抗原性相关产物在其组装过程中与外致密纤维相关联。

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