Research on p85a Gene in Radial Migration of Cortical Projection Neurons of the Mouse Cortex

Abstract: In recent years， Pi3k/Akt signaling pathway has been found vitally in cell multiplication and regulation. Activation of Pi3k can modulate a variety of cellular functions. Pi3k consists of p85a and p110. In this study， we aimed to determine whether p85a regulates the early migration of cortical projection neurons in mice by interruption the expression of p85a. First， we designed a scrambled shRNA plasmid as a control， two short hairpin RNAs (sip85a-1， sip85a-2) construct against p85a and an overexpression plasmid (OEp85a); Constructs were tested by transiently transfecting N2a cells. We detected the p85a mRNA level by Real-time PCR analysis. Then， we used two effective shRNAs to determine whether p85a was involved in the migration of cortical neurons. Real-time PCR analysis showed that p85a mRNA level decreased significantly after transfection with the plasmids express ing sip85a-1 (40%) and sip85a-2 (40%) compared to control (P<0.05). Conversely， p85a mRNA level increased significantly after transfection with OEp85a (P<0.01). In order to quantify in utero electroporation results， we also counted EGFP-positive cells in different bins. The results showed that all of sip85a-1， sip85a-2 and OEp85a could impair radial migration significantly (P<0.05). These observations indicate that balance of p85a has vital functions in regulating migration of cortical neurons during brain development.

CSTR: 32200.14.cjcb.2014.08.0003