Andonian S, Hermo L
Department of Anatomy and Cell Biology, McGill University, Montréal, Québec, Canada.
J Androl. 1999 May-Jun;20(3):415-29.
The epithelial cells lining the cauda epididymidis and vas deferens are active in endocytosis and have an abundance of lysosomes and a well-characterized secretory apparatus. However, little is known about the nature of lysosomal proteins contained within lysosomes, the types of receptors on the cell surface, and the types of proteins secreted by these cells. In the present study, cathepsins A, D, B, and sulfated glycoprotein (SGP)-1, well-characterized lysosomal proteins, as well as SGP-2, a secretory protein and low-density lipoprotein receptor-related protein-2 (LRP-2), an endocytic receptor, were immunolocalized at the light-microscopic level within epithelial cells of the cauda epididymidis and vas deferens. Principal cells showed numerous intensely reactive lysosomes for cathepsins A, D, and SGP-1 in all regions of the cauda and vas deferens and for cathepsin B only in the cauda epididymidis. Basal cells were intensely reactive for cathepsin A, unreactive for cathepsins D and B, and weakly reactive for SGP-1 in the cauda region. In the vas deferens, these cells were intensely reactive for cathepsin A and SGP-1 and unreactive for cathepsin B; in the case of cathepsin D, basal cells were weakly reactive in the proximal vas deferens but intensely reactive in the middle and distal vas deferens. Clear cells, present in the cauda region and proximal vas deferens, were intensely reactive for cathepsin A, weakly reactive for SGP-1, and unreactive for cathepsins D and B, while narrow cells found mainly in the proximal vas deferens were intensely reactive for cathepsins A, D, and SGP-1 and unreactive for cathepsin B. Thus, the expression of different lysosomal enzymes in the cauda epididymidis and vas deferens is not only cell- but also region-specific, suggesting differences in the type of substrates internalized by these cells. SGP-2, a secretory protein, showed a checkerboardlike staining pattern in the cytoplasm of principal cells of the cauda epididymidis, while the cytoplasm of all principal cells were intensely reactive in the vas deferens. This type of reaction, as well as staining of sperm, suggests that SGP-2 is secreted into the lumen, where it functions in relation to sperm. The endocytic receptor LRP-2 was noted only on the apical surface of principal cells of the cauda and vas deferens and in spherical structures indicative of endosomes suggestive of their role in the uptake of various ligands, including SGP-2, for which it has a high binding affinity. Thus SGP-2 in the cauda and vas deferens is not only secreted but endocytosed by principal cells, suggestive of an active turnover in the lumen. In summary, the epithelial cells of the cauda and vas deferens show marked differences in expression of lysosomal proteins, SGP-2, and LRP-2 suggestive of differences in their functional activity while sperm are stored and protected in these regions.
附睾尾部和输精管内衬的上皮细胞具有活跃的内吞作用,有大量溶酶体和特征明确的分泌装置。然而,对于溶酶体内所含溶酶体蛋白的性质、细胞表面受体的类型以及这些细胞分泌的蛋白类型,人们了解甚少。在本研究中,组织蛋白酶A、D、B和硫酸化糖蛋白(SGP)-1(特征明确的溶酶体蛋白)以及分泌蛋白SGP-2和内吞受体低密度脂蛋白受体相关蛋白-2(LRP-2),在光镜水平上被免疫定位到附睾尾部和输精管的上皮细胞内。主细胞在附睾尾部和输精管的所有区域对组织蛋白酶A、D和SGP-1显示出大量强烈反应性的溶酶体,而仅在附睾尾部对组织蛋白酶B有反应。基底细胞在附睾尾部区域对组织蛋白酶A有强烈反应,对组织蛋白酶D和B无反应,对SGP-1反应较弱。在输精管中,这些细胞对组织蛋白酶A和SGP-1有强烈反应,对组织蛋白酶B无反应;对于组织蛋白酶D,基底细胞在输精管近端反应较弱,但在输精管中部和远端反应强烈。存在于附睾尾部区域和输精管近端的透明细胞对组织蛋白酶A有强烈反应,对SGP-1反应较弱,对组织蛋白酶D和B无反应,而主要位于输精管近端的窄细胞对组织蛋白酶A、D和SGP-1有强烈反应,对组织蛋白酶B无反应。因此,附睾尾部和输精管中不同溶酶体酶的表达不仅具有细胞特异性,还具有区域特异性,这表明这些细胞内化的底物类型存在差异。分泌蛋白SGP-2在附睾尾部主细胞的细胞质中呈现棋盘状染色模式,而在输精管中所有主细胞的细胞质都有强烈反应。这种反应类型以及精子的染色表明,SGP-2分泌到管腔中,在那里它与精子发挥相关功能。内吞受体LRP-2仅在附睾尾部和输精管主细胞的顶端表面以及指示内体的球形结构中被发现,这表明它们在摄取包括SGP-2在内的各种配体中发挥作用,LRP-2对SGP-2具有高结合亲和力。因此,附睾尾部和输精管中的SGP-2不仅被主细胞分泌,还被其内化,这表明管腔内存在活跃的周转。总之,附睾尾部和输精管的上皮细胞在溶酶体蛋白、SGP-2和LRP-2的表达上存在显著差异,这表明在精子储存和保护于这些区域时,它们的功能活性存在差异。