αvβ5 integrin mediates the effect of vitronectin on the initial stage of differentiation in mouse cerebellar granule cell precursors

Abstract

Vitronectin (VN), one of the extracellular matrix proteins, controls the maturation of cerebellar granule cells (CGCs) through the promotion of the initial differentiation stage progress. However, the receptors of VN in the initial differentiation stage of CGC precursors (CGCPs) have not been clarified. In this study, we characterized the receptor candidates for VN in CGCPs. First, we confirmed that αvβ3 and αvβ5 integrins, which are receptor candidates for VN, were co-localized with VN in the developing cerebellum and primary cultured CGCPs. Next, the knockdown (KD) of αv, β3, and β5 integrins with small interference RNA (siRNA) for each integrin reduced the ratio of Tuj1, a final differentiation marker, -positive CGCPs. We further studied whether αvβ3 and αvβ5 integrins control the initial differentiation stage. The KD of αv and β5, but not β3, integrins significantly increased the ratio of transient axonal glycoprotein 1 (TAG1), an initial differentiation marker, -positive CGCPs, whereas the KD of αv and β3 integrins, not β5 integrin, stimulated the proliferation of CGCPs. Overexpression of β5 integrin stimulated the progress of the initial differentiation stage as well. To confirm the interaction between αvβ5 integrin and VN, VN was added to β5 integrin-KD CGCPs. The promotion of the progress of initial differentiation by VN was abrogated by β5 integrin KD using small hairpin RNA (shRNA). Taken together, our results indicated that αvβ5 integrin, as the very receptor of VN, is responsible for the progress of the initial differentiation stage in mouse CGCPs.

Keywords: Cerebellar granule cell; Differentiation; Vitronectin; αvβ3 integrin; αvβ5 integrin.