Mechanisms Of GABA/GABA-A Receptor Pathway Inhibiting Migration Function Of Dendritic Cells In Mice

Objective:To investigate the mechanism of GABA/GABA-A receptor pathway inhibiting migration function of dendritic cell in mice.Method:Forty-eight hours after transfection of GABA-A receptor α5 subunit SiRNA in immature dendritic cells(iDCs),the expression of GABA-A receptor α5 subunit was detected using Western Blot(WB).Then,the transwell migration experiment was performed to assess the regulation of iDCs migration ability by GABA/GABA-A receptor pathway.The regulatory effects of GABA/GABA-A receptor pathway on iDCs pseudopodia production was further analyzed using laser confocal microscopy.Immunofluorescence and flow cytometry were employed to examine the distribution of the GABA-A receptor α5 subunit inside and outside the iDCs membrane,and to assess the internalization and recycling of the GABA-A receptor α5 subunit under the action of GABA.Co-immunoprecipitation was performed to determine whether GABA-A receptor α5 subunits,radixin,and F-actin formed complexes under the action of GABA.WB was used to investigate the regulation of p38 MAPK pathway by GABA/GABA-A receptor pathway and its effect on the expression of phospho-cofilin.Result:1.Forty-eight hours after the transfection of GABA-A receptor α5 subunit SiRNA in iDCs,the expression of α5 subunit significantly decreased(p<0.05).2.The migration of iDCs was significantly inhibited by GABA(p<0.05).3.GABA inhibited the formation of iDCs pseudofoot.4.The GABA-A receptor α5 subunit was internalized under the action of GABA(p<0.05).This effect lasted up to 3 hours,after which the internalizedα5 subunit gradually circulated back to the membrane.5.Under the action of GABA,the GABA-A receptor α5 subunit internalizeeds with radixin and F-actin to form the immune complexes..6.The GABA/GABA-A receptor pathway induced the expression of phospho-cofilin(p<0.05)probably by inhibiting the p38 MAPK pathway(p<0.05).Conclusion:GABA/GABA-A receptor pathway inhibits the migration ability of iDCs,which increases the expression of phospho-cofilin through the inhibition of the p38 MAPK pathway by forming immune complexes.