β-PIX-d, a Member of the ARHGEF7 Guanine Nucleotide Exchange Factor Family, Activates Rac1 and Induces Neuritogenesis in Primary Cortical Neurons.

Abstract

β-PIX, a Rac1/Cdc42-specific guanine nucleotide exchange factor, is known to regulate actin cytoskeleton remodeling during cell migration. In this study, we investigated the effects of β-PIX-d, an isoform of β-PIX, on neocortical development and neuritogenesis. Overexpression of β-PIX-d in the embryonic neocortex induced increased cell clusters and enhanced neurite outgrowth in cortical neurons. Following in utero electroporation of β-PIX-d expression vectors into neuronal progenitor cells at embryonic day 13.5 (E13.5), histological analysis at postnatal day 0 (P0) revealed the presence of clustered neurons and neurites outside of the marginal zone (MZ). Immunofluorescence staining with the neuronal marker TuJ1 confirmed that the clustered structures were predominantly composed of neurons. Layer-specific marker analysis further demonstrated the misplacement of layer V-VI neurons into layer I and the subarachnoid space. In primary neocortical cultures, β-PIX-d overexpression promoted neuritogenesis and increased Rac1 activity, as detected by pull-down assays. These findings suggest that β-PIX-d and Rac1 interactions play a critical role in the formation of neocortical clustering and the regulation of neuritogenesis.