Three-dimensional organization of the developing vasculature of the kidney.

Kidney function depends on a well-developed vascular system. Any impairment of the blood supply disturbs the integrity and function of the organ. The differentiation of renal vessels has been investigation for many years, but little is known about the relationship between nephrogenesis and vessel development. In the present work the spatial organization of the differentiating vessels was analyzed in precisely oriented tissue sections and in optical sections acquired by laser scan microscopy. Developing vessels as well as small capillaries were visualized with two endothelium-detecting antibodies. Small vessels running in parallel towards the organ capsule were detected in numerous cortico-medullary-oriented tissue sections. Cross-sections of the nephrogenic zone showed a regularly arranged network, which was composed of cells detected by both monoclonal antibodies. Parts of this network were localized in regions of the nephrogenic zone which have been assumed to be free of vessels or vessel-like structures for a long time. These results were confirmed by the laser-scan-microscopic analysis of complete cortex explants. The extraordinarily regular arrangement of the endothelial network in the nephrogenic zone allowed us to reconstruct the developing vascular system. The results presented here underline the close relationship between nephrogenesis and vessel development.

Stimulation of renal microvascular development under organotypic culture conditions.

The development of the renal vascular system requires the coordinated action of soluble morphogenic factors and specific extracellular matrix components. Despite intensive research it remains unknown whether the humoral or the environmental component is more important in the development of renal microvessels. The prolonged serum-free culture of embryonic kidney cortex explants was achieved by means of a newly developed perfusion culture system. This system made the investigation of renal vascular development under defined organotypic conditions possible. Thus, growth factors such as vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and hormones (aldosterone, vitamin D3) could be applied without the interference with serum components. Medium supplementation with VEGF or aldosterone in combination with vitamin D3 resulted in the coordinated proliferation of endothelial cells in the explant. A well-developed collecting duct epithelium and numerous tubular structures were always observed. In contrast, only a uniform cell layer was found between fibrous organ capsule and the collecting duct epithelium after bFGF application, but neither tubular structures nor endothelial cells. Thus, the experiments indicate that bFGF alone has no stimulating effect on the growth of the renal microvasculature under perfusion culture conditions.