Abou-Haila A, Tulsiani D R, Skudlarek M D, Orgebin-Crist M C
Laboratoire de Biologie Cellulaire, Université René Descartes, Paris, France.
J Androl. 1996 May-Jun;17(3):194-207.
The expression and androgen regulation of beta-glucuronidase molecular forms were examined in mouse epididymis, liver, and kidney. Two-dimensional polyacrylamide gel electrophoresis performed under nondenaturing conditions showed that, compared to liver and kidney, which contain four microsomal (M1-M4) and a major lysosomal (L) form of beta-glucuronidase, the epididymis revealed regional differences and tissue specificity in the expression of the various molecular forms of the enzyme. Only the lysosomal form (pI 5.4-6.1) is present in the caput epididymidis while the corpus/cauda contains the lysosomal form, the free X form (pI 5.9-6.3) and the four microsomal forms (X form complexed with egasyn). Mutant mice that lack egasyn have no microsomal forms in the distal epididymis. In epididymal fluid, the lysosomal form is found throughout the epididymis, whereas the X form appears only in the corpus/cauda epididymidis. Sodium dodecyl Sulfate (SDS)-gel electrophoresis and western blot analysis of immunoprecipitated beta-glucuronidase revealed only one band corresponding to the L form (apparent molecular weight 74 kDa) in the caput epididymidis and two bands in the corpus/cauda (apparent molecular weights 73 and 75 kDa), corresponding to L and X forms, respectively. Castration of mice led to the suppression of the regional differences in the appearance of X and M forms in the epididymis. Testosterone supplementation to castrated mice restored the characteristic electrophoretic pattern of beta-glucuronidase in the caput epididymidis. In the liver and kidney, castration has no effect on the expression of the molecular forms, whereas androgen treatment induced the X form in the kidney. Histochemical localization of beta-glucuronidase confirmed the region specificity seen in the epididymis and in addition revealed cell specificity in the expression of beta-glucuronidase. These results indicate that beta-glucuronidase shows tissue specificity and, in the case of the epididymis, region and cell specificity. In addition, the enzyme in the different tissues responds differentially to androgens.
在小鼠附睾、肝脏和肾脏中检测了β-葡萄糖醛酸酶分子形式的表达及雄激素调节情况。在非变性条件下进行的二维聚丙烯酰胺凝胶电泳显示,与含有四种微粒体(M1 - M4)和一种主要溶酶体(L)形式β-葡萄糖醛酸酶的肝脏和肾脏相比,附睾在该酶各种分子形式的表达上表现出区域差异和组织特异性。附睾头仅存在溶酶体形式(pI 5.4 - 6.1),而附睾体/尾含有溶酶体形式、游离X形式(pI 5.9 - 6.3)和四种微粒体形式(与egasyn复合的X形式)。缺乏egasyn的突变小鼠在附睾远端没有微粒体形式。在附睾液中,溶酶体形式在整个附睾中都有发现,而X形式仅出现在附睾体/尾。十二烷基硫酸钠(SDS)凝胶电泳和免疫沉淀的β-葡萄糖醛酸酶的蛋白质印迹分析显示,附睾头只有一条对应L形式(表观分子量74 kDa)的条带,而附睾体/尾有两条条带(表观分子量分别为73和75 kDa),分别对应L和X形式。小鼠去势导致附睾中X和M形式出现的区域差异受到抑制。给去势小鼠补充睾酮可恢复附睾头β-葡萄糖醛酸酶的特征性电泳图谱。在肝脏和肾脏中,去势对分子形式的表达没有影响,而雄激素处理可诱导肾脏中出现X形式。β-葡萄糖醛酸酶的组织化学定位证实了附睾中所见的区域特异性,此外还揭示了β-葡萄糖醛酸酶表达的细胞特异性。这些结果表明,β-葡萄糖醛酸酶表现出组织特异性,在附睾的情况下,还表现出区域和细胞特异性。此外,不同组织中的该酶对雄激素的反应不同。