Ultrastructural localization of proteoglycans by cationic dyes in the epithelial-stromal interface of the guinea pig lateral prostate

Abstract

Proteoglycans (PGs) in the epithelial-stromal interface of the guinea pig lateral prostate were localized at ultrastructural level, using cuprolinic Blue (CB), alcian Blue (AB), and ruthenium red (RR). After staining with CB or AB according to the critical electrolyte concentration method (CEC), PGs appeared as short electron-dense filaments. According to their sizes and location, three type (T1, T2, T3) of CB-stained filaments were identified. T1 filaments were short (25 nm) and were found on both sides of the lamina densa of the basal laminae of the prostatic epithelium, smooth muscle cells, and capillary endothelial cells. They were regularly spaced with an interval of 60 nm. T1 filaments were more randomly distributed in the lamina densa. T2 CB filaments were ~30-40 nm long and closely associated with the collagen fibrils. They were usually arranged perpendicular to the long axis of collagen fibrils also at intervals of 60 nm. T3 filaments were found in different regions of the lamina propria, including: 1) reticular layer (pars fibroreticularis) below the basal lamina; 2) interstitial spaces; 3) closely associated with the cell surfaces of fibroblasts; and 4) around the collagen fibrils. Their sizes were variable (60-100 nm) and more densely stained. AB revealed similar patterns of PG distribution, except that the three types of PG filaments were longer but thinner. When the tissues were stained with RR, or RR-AB combined. PGs appeared as dense granules of various sizes, instead of filaments. Their locations and distributions were similar to those of the CB filaments, except that in the case of combined RR-AB treatment the PG granules were linked by a fine filamentous network, suggesting the interconnecting nature of the PGs and other extracellular components.