The ultrastructural localization of the enzymes related to steroid hormone metabolism in the guinea-pig testis

Abstract

A study of the ultrastructural localization of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD), glucose-6-phosphate dehydrogenase (G-6-PD), beta-hydroxybutyrate dehydrogenase (beta-HBD), NADH diaphorase (NADH-D) and NADPH diaphorase (NADPH-D) in the guinea-pig testis is reported. The procedures employed included short immersion or perfusion fixation with aldehydes followed by incubation of small blocks in a tetrazolium salt or a ferricyanide medium. The effects of incubation conditions were investigated, and a reaction medium for the ultracytochemical demonstration of 11 beta-HSD is described. Using suitable controls, evidence for the specificity of the cytochemical reactions is presented. It was found that all the enzymes studied were present in both the Leydig and Sertoli cells of the guinea-pig testis and that the intracellular distribution pattern for each enzyme was independent of the cell type. Using tetrazolium salt techniques, both 3 beta-HSD and 11 beta-HSD activities were localized on or in membranes of smooth endoplasmic reticulum and within the mitochondria. With the ferricyanide techniques, G-6-PD activity was found to be associated mainly with the smooth endoplasmic reticulum membranes, while beta-HBD activity was limited to mitochondria. With both the tetrazolium salt and ferricyanide techniques, the reaction products for NADH-D and NADPH-D activities showed localizations which were similar to those observed for the steroid dehydrogenases.