Ultrastructural evidence for morphological specificity in isolated bovine retinal capillary basement membranes

Because of its relative availability, large size, and presumed similarity to human, the bovine retina has been used by numerous investigators as a source of vessels, cells, and basement membranes (BMs) for biochemical analyses and in vitro studies of cells and extracellular matrix. Careful morphological studies of these vessels and their associated BMs, however, have not been done. Accordingly, we carried out experimental ultrastructural studies in an effort to show their cellular composition, their histoarchitectural relationships within retinal capillary walls, and the disposition and features of their isolated BMs. Our study shows that these vessels are complex, multicomponent structures composed of endothelial cells and intramural pericytes, which frequently communicate via direct cell/cell contacts, and a system of BMs. The latter includes continuous Muller cell BMs, interrupted subendothelial BMs, and pericytic BMs with masses of pericytic matrix (PCM) intervening. Isolated subendothelial BMs are remarkable for fenestrations, selective susceptibility to nonspecific proteases, and high density of ruthenium red (RR)-positive anionic sites. On the contrary, Muller cell BMs are continuous (completely surrounding retinal capillaries), relatively refractory to proteases, and show significantly fewer anionic sites by RR. Acellular capillary BMs frequently show ghost-like "pockets" previously occupied by pericytes. These are surrounded by pericytic BMs and interstitial spaces are "filled-in" by a BM-like material (PCM) which frequently contains striated collagen fibrils and is positionally and morphologically homologous to glomerular mesangial matrix. These data indicate that tissue specificity of BMs may be far more precise than previously thought and that each capillary BM leaflet may possess a peculiar macromolecular architecture commensurate with its specific function.