Hermo L, Wright J, Oko R, Morales C R
Department of Anatomy, McGill University, Montreal, Quebec, Canada.
Biol Reprod. 1991 Jun;44(6):1113-31. doi: 10.1095/biolreprod44.6.1113.
The localization of sulfated glycoprotein-2 (clusterin; SGP-2) was investigated in the rete testis, efferent ducts, and epididymis of the rat using light (LM) and electron (EM) microscope immunocytochemistry. At the LM level, the epithelial cells of the rete testis and efferent ducts demonstrated an intense immunoperoxidase reaction over their apical and supranuclear regions, and sperm in the lumen of the efferent ducts were unreactive. In the EM, gold particles were found exclusively over the endocytic apparatus of these cells. In the proximal area of the epididymal initial segment, an insignificant immunostaining of epithelial cells and sperm was observed. However, the distal area of the initial segment showed a moderate staining over the epithelial principal cells and sperm, while in the intermediate zone of the epididymis a stronger reaction was observed over these cells. The strongest immunoperoxidase reaction was noted in the caput epididymidis, where it formed a distinct mottled pattern. Thus, while some principal cells were intensely stained, others were moderately or weakly stained; a few were completely unreactive. In the corpus and cauda epididymidis, the staining pattern was similar but not as intense. In the EM, only the secretory apparatus of these cells was found to be immunolabeled with gold particles. Sperm in the lumen of these different regions were also labeled. The epithelial clear cells were unreactive throughout the epididymis. Northern blot analysis substantiated these results and showed the presence of highest levels of SGP-2 mRNA in the caput epididymidis, especially in its proximal area, whereas increasingly lower levels were found in the corpus and cauda epididymidis. In summary, these results suggest that testicular SGP-2 dissociates from the sperm during passage through the rete testis and efferent ducts, where it is endocytosed by the epithelial cells lining these regions. In the epididymis, it is replaced by an epididymal SGP-2 that is secreted by the epithelial principal cells of the epididymis. Furthermore, in the epididymis, the principal cells appear to be in different functional states with respect to the secretion of epididymal SGP-2 within a given region of the duct as well as along the epididymal duct.
运用光镜和电镜免疫细胞化学技术,对大鼠睾丸网、输出小管及附睾中硫酸化糖蛋白-2(聚集素;SGP-2)的定位进行了研究。在光镜水平,睾丸网和输出小管的上皮细胞在其顶端和核上区域呈现强烈的免疫过氧化物酶反应,而输出小管管腔内的精子无反应。在电镜下,仅在这些细胞的内吞装置上发现金颗粒。在附睾起始段的近端区域,上皮细胞和精子的免疫染色不明显。然而,起始段的远端区域在上皮主细胞和精子上显示中度染色,而在附睾中间区,这些细胞上观察到更强的反应。在附睾头观察到最强的免疫过氧化物酶反应,形成明显的斑驳图案。因此,虽然一些主细胞染色强烈,但其他细胞染色中度或轻度;少数细胞完全无反应。在附睾体和附睾尾,染色模式相似但不那么强烈。在电镜下,仅发现这些细胞的分泌装置被金颗粒免疫标记。这些不同区域管腔内的精子也被标记。附睾中的清亮细胞在整个附睾中均无反应。Northern印迹分析证实了这些结果,并显示附睾头中SGP-2 mRNA水平最高,尤其是在其近端区域,而在附睾体和附睾尾中水平逐渐降低。总之,这些结果表明,睾丸SGP-2在通过睾丸网和输出小管的过程中与精子分离,在此处被这些区域的内衬上皮细胞内吞。在附睾中,它被附睾上皮主细胞分泌的附睾SGP-2所取代。此外,在附睾中,主细胞在附睾管的给定区域内以及沿附睾管分泌附睾SGP-2方面似乎处于不同的功能状态。