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Vitronectin promotes the progress of the initial differentiation stage in cerebellar granule cells

Journal Reference

Mol Cell Neurosci. 2016 Jan;70:76-85.

Hashimoto K1, Sakane F1, Ikeda N1, Akiyama A1, Sugahara M1, Miyamoto Y2.

Show Affiliations
  1. Graduate School of Humanities and Sciences, Ochanomizu University, Otsuka 2-1-1, Bunkyo-ku, Tokyo 112-8610, Japan.
  2. Graduate School of Humanities and Sciences, Ochanomizu University, Otsuka 2-1-1, Bunkyo-ku, Tokyo 112-8610, Japan. Electronic address: [email protected]

Abstract

Vitronectin (VN), which is an extracellular matrix protein, is known to be involved in the proliferation and differentiation of primary cultured cerebellar granule cell precursors (CGCPs); however, the effect of Vitronectin is not fully understood. In this study, we analyzed the effects of Vitronectin loss on the proliferation and differentiation of CGCPs in Vitronectin knockout (VNKO) mice in vivo. First, immunohistochemistry showed that Vitronectin was distributed in the region from the inner external granule layer (iEGL) through the internal granule layer (IGL) in wild-type (WT) mice. Next, we observed the formation of the cerebellar cortex using sagittal sections of VNKO mice at postnatal days (P) 5, 8 and 11. Loss of Vitronectin suppressed the ratio of NeuN, a neuronal differentiation marker, to positive   cerebellar granule cells (CGCs) in the external granule layer (EGL) and the ratio of CGCs in the IGL at P8, indicating that the loss of Vitronectin suppresses the differentiation into CGCs. However, the loss of Vitronectin did not significantly affect the proliferation of CGCPs. Next, the effect of Vitronectin loss on the initial differentiation stage of CGCPs was examined. The loss of Vitronectin increased the expression levels of Transient axonal glycoprotein 1 (TAG1), a marker of neurons in the initial differentiation stage, in the cerebella of VNKO mice at P5 and 8 and increased the ratio of TAG1-positive cells in the primary culture of VNKO-derived CGCPs, indicating that the loss of VN accumulates the CGCPs in the initial differentiation stage. Taken together, these results demonstrate that Vitronectin promotes the progress of the initial differentiation stage of CGCPs.

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