Sonic Hedgehog Mediator Gli2 Regulates Bladder Mesenchymal Patterning

Abstract

Congenital bladder anomalies are a major challenge to pediatric urologists. Understanding the mechanism of bladder development is crucial for advancing patient treatment. Current evidence suggests that Shh (R&D Systems) is an epithelial signal regulating bladder development, although the mechanism of the regulation is still unclear. We examined the regulation of bladder mesenchymal development. Mutation analysis, immunohistochemistry, immunoblot, in situ hybridization, and primary cell culture and transfection were performed. The mesenchyma proximal to the epithelium was defined as the inner zone and that distal to the epithelium was defined as the outer zone. We found that the Shh transcriptional factor Gli2 and the Shh target gene Bmp4 (R&D Systems) were expressed in the inner mesenchymal zone of the bladder, where active cell proliferation was observed. In Gli2(-/-) bladder primary mesenchymal cell cultures transfection with adenoviruses expressing DeltaNGli2, a constitutionally active form of Gli2, up-regulated Bmp4 expression and promoted cell proliferation. In the outer mesenchymal zone, where Gli2 and Bmp4 expression was not detectable, smooth muscle alpha-actin was expressed. In Gli2(-/-) embryo bladders Bmp4 expression in the inner zone was lost and ectopic smooth muscle was detected in the inner mesenchymal zone. Exogenous Bmp4 (10 ng/ml) in primary smooth muscle cell culture repressed smooth muscle differentiation and repression was partially rescued by the Bmp4 antagonist Noggin (R&D Systems) (300 ng/ml). Our data suggests that the Shh transcriptional factor Gli2 regulates radial patterning of the bladder mesenchyma.