Visualization of Golgi complexes and spermatogonial cohorts of viable, intact seminiferous tubules.

Golgi complexes of cells within the intact, viable seminiferous tubule were examined by light microscopy using the vital Golgi stain C6-NBD-ceramide. This staining technique provided a quick and simple method to visualize all Golgi complexes of cells within the seminiferous tubule that are directly accessible to the basal compartment. Thus, peritubular, spermatogonial, and Sertoli cell Golgi complexes were visualized. Peritubular cells contained simple Golgi complexes which did not change with the stage of the seminiferous epithelium. Both solitary spermatogonial Golgi complexes which varied in size and number with the stage of the seminiferous epithelium and cohorts of spermatogonia connected by intercellular bridges were easily visualized. The Golgi complexes of Sertoli cells were located in the basal, perinuclear cytoplasm except in Stages VII-VIII, when they extended towards the lumen. Exposure of isolated seminiferous tubules to the fungal metabolite brefeldin A caused the Sertoli cell Golgi complex staining pattern to become diffuse or to co-localize with heads of elongate spermatids. The Golgi complexes of the peritubular cells and spermatogonia were resistant to brefeldin A. The C6-NBD-ceramide vital staining method should be useful for studying stage-dependent Sertoli cell Golgi complex movement and spermatogonial maturation.