The Function Of Cotl1 In Mouse Cortical Development

The development of mammalian cerebral cortex includes the proliferation,differentiation of cells and the neurons migrate to the correct location in the cortex,eventually forming a highly organized 6-layered structure.Neuronal migration requires the remodeling and regulation of the cytoskeleton and the adhesion between cells.The abnormal of neuronal migration will cause malformations of cerebral cortical and lead to neuroogical diseases,such as alzheimer's disease and epilepsy.F-actin is involved in the remodeling maintained the stability of cytoskeleton.Cotl1,a member of the actin depolymeration factor,Mainly with cofilin competitively binding F-actin.The New research shows that,cofilin cleaves F-actin to produce new fibrous ends,so that the F-actin can extend.Therefore,cofilin is involved in maintaining the dynamic remodeling of the cytoskeleton,guiding cell migration and plays an important role in the development of the cerebral cortex.However,there are few studies on the stability of Cotl1 in the cytoskeleton and the cell motility.To investigate the effect of Cotl1 on cerebral cortical,this study constructed the vector of overexpression and the point mutation.Using in utero electroporation and immunofluorescence to study the function of Cotl1 in neuronal migration,cell proliferation and differentiation.The main results are as follows:1.In this study,the Cotl1 gene fragment was successfully cloned;Using semi-quantitative approach,we detected the expression of Cotl1 in the embryonic mouse cerebral cortex at the specific time.2.We successfully constructed the Cotl1 overexpression plasmid pCAG-Cotl1-myc;Using in utero electroporation,we overexpressed Cotl1 in the developing mouse cortial early-generate neurons at E12.5 and found that overexpression of Cotl1 in a dose-dependent manner inhibited the early neuronal migration.Overexpressed Cotl1 in the developing mouse cortial late-generate neurons at E15.5 delayed the arrival time of neurons to the CP and the leading process were significantly increased compared with the control group at high magnification.3.Using in utero electroporation and the BrdU labering technology to study the effect of cell proliferation and found that overpression of the Cotl1 didn't affect the proliferation;Overpression of this plasmid didn't alter the differentation of neonatal cells into neurons.4.Using point mutation,we successfully constructed the Cotl1-ABM plasmid pCAG-Cotl1-ABM-myc;We overexpressed pCAG-Cotl1-ABM-myc in the mouse cortical neurons at E15.5 and found that the neuronal migration and the length of leading process returned to normal.These results indicate that overexpression of Cotl1 inhibits the migration of neurons but has no significant effect on the proliferation and differentiation of neurons in the cerebral cortex.And the sites of 73 lysine and 75 arginine play an important role in neuronal migration.This study provides a theoretical basis for the research of neurological diseases.