Coexpression of neuropilin-1, Flk1, and VEGF164in developing and mature mouse kidney glomeruli

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Abstract
Neuropilin-1, a neuronal cell surface semaphorin III receptor protein important for axonal guidance in developing peripheral nervous system efferents, has also been identified as a vascular endothelial growth factor (VEGF) receptor on endothelial cells. To evaluate its expression in kidney, we carried out RT-PCR on newborn and adult total renal RNAs. A 403-bp product, which was predicted to be that from neuropilin-1 mRNA, was found in both samples. Nucleotide sequencing confirmed that these products encoded neuropilin-1. Northern analysis of newborn and adult kidney RNA showed specific hybridization to appropriately sized bands of ∼6 kb. In situ hybridization with a mouse-specific antisense neuropilin-135S-cRNA probe showed distinct glomerular localization on sections from both newborns and adults. Similar patterns of hybridization were seen in sections treated with antisense cRNA probes against another VEGF receptor, Flk1, and with VEGF probes. However, the VEGF hybridization signal was markedly less in adult glomeruli than those for neuropilin-1 and Flk1. Because neuropilin-1 specifically binds VEGF165in humans, we carried out RT-PCR on mouse kidney RNA with primers that amplified the three alternatively spliced isoforms of VEGF mRNA. Our analysis showed that for both newborn and adult kidneys, the relative abundance of VEGF mRNA was VEGF164≫ VEGF120> VEGF188. We conclude that the expression of neuropilin-1, in conjunction with Flk1 and VEGF164, jointly contributes to the development and maintenance of glomerular capillaries.