Igdoura S A, Hermo L, Morales C R
Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, Canada.
Microsc Res Tech. 1994 Dec 15;29(6):468-80. doi: 10.1002/jemt.1070290605.
The epithelial nonciliated cells of the efferent ducts are specialized in internalizing many luminal substances. The nonciliated cells actively endocytose sulfated glycoprotein-2 (SGP-2), a major secretory protein of Sertoli cells and a homologue of human apolipoprotein J. This study was undertaken to investigate the internalization of Sertoli-derived SGP-2 and synthesis of an endogenous efferent duct form of SGP-2 by nonciliated cells targeted to their secondary lysosomes on animals whose efferent ducts were ligated and/or received injections of tunicamycin. The regulation of synthesis of the endogenous form of SGP-2 within nonciliated cells by hormones in general and testosterone in particular was also examined using hypophysectomized and castrated animals with or without subsequent testosterone replacement. Quantitative electron microscope immunocytochemistry was performed on groups of animals fixed with 4% paraformaldehyde and 0.5% glutaraldehyde in phosphate buffer for each experimental condition and their controls. In each case, the labeling density (number of gold particles/micron 2) within the endosomal (endosomes) and lysosomal (dense multivesicular bodies and secondary lysosomes) compartments was calculated. The results revealed that ligation of the efferent ducts resulted in a significant decrease in the labeling density of the endosomal and lysosomal compartments. However, a baseline of about 18% of controls was still observed in the lysosomal compartment 24 h after ligation. In this compartment similar values were noted 24 h after tunicamycin treatment in conjunction with or without ligation. These results suggest that an endogenous form of SGP-2 is synthesized by nonciliated cells and presumably targeted via small vesicles from the Golgi apparatus to the lysosomal compartment, but that the major portion of SGP-2 within this compartment is derived via endocytosis of testicular SGP-2. Hypophysectomy and castration also showed significant decreases in the labeling densities of these two compartments, but again a baseline level of labeling was noted in the lysosomal compartment. Subsequent testosterone administration to 7-day hypophysectomized or castrated animals had no effect on the labeling density of the lysosomal compartment, as values comparable to the effect of hypophysectomy or castration alone were noted. Taken together these results suggest that the nonciliated cells of the efferent ducts synthesize an endogenous form of SGP-2 that is targeted to the lysosomal compartment and which is not regulated by pituitary factors or testosterone.
输出小管的上皮无纤毛细胞专门负责内化许多管腔物质。无纤毛细胞能主动内吞硫酸化糖蛋白-2(SGP-2),它是支持细胞的一种主要分泌蛋白,也是人类载脂蛋白J的同源物。本研究旨在调查支持细胞来源的SGP-2的内化情况,以及在输出小管被结扎和/或接受衣霉素注射的动物中,无纤毛细胞将内源性输出小管形式的SGP-2合成并靶向运输至其次级溶酶体的情况。还使用了垂体切除和阉割的动物,在有或没有后续睾酮替代的情况下,研究了激素尤其是睾酮对无纤毛细胞内源性SGP-2合成的调节作用。对每组在每种实验条件及其对照下用磷酸盐缓冲液中的4%多聚甲醛和0.5%戊二醛固定的动物进行定量电子显微镜免疫细胞化学分析。在每种情况下,计算内体(内体)和溶酶体(致密多囊泡体和次级溶酶体)区室内的标记密度(金颗粒数/微米²)。结果显示,输出小管结扎导致内体和溶酶体区室的标记密度显著降低。然而,结扎后24小时,溶酶体区室中仍观察到约为对照值18%的基线水平。在该区域,衣霉素处理结合或不结合结扎24小时后也观察到类似的值。这些结果表明,内源性SGP-2由无纤毛细胞合成,推测是通过来自高尔基体的小泡靶向运输至溶酶体区室,但该区域内SGP-2的主要部分是通过睾丸SGP-2的内吞作用获得的。垂体切除和阉割也显示这两个区室的标记密度显著降低,但溶酶体区室中再次观察到标记的基线水平。对垂体切除或阉割7天的动物随后给予睾酮,对溶酶体区室的标记密度没有影响,因为观察到的值与单独垂体切除或阉割的效果相当。综合这些结果表明,输出小管的无纤毛细胞合成一种内源性SGP-2,其靶向运输至溶酶体区室,且不受垂体因子或睾酮的调节。