THE EFFECTS OF OSMOTIC PRESSURE CHANGING ON CONNEXINS EXPRESSION IN CULTURED RAT HYPOTHALAMIC ASTROCYTES AND NEURONS

Objective To investigate the effects of osmotic pressure changing on connexin43 (Cx43) and Cx32 expression in cultured rat hypothalamic astrocytes and neurons. Methods By culturing astrocytes and neurons from newborn rats,these cells were divided to two groups.The cells in first group were moved from isotonic culture medium to hypertonic culture medium and incubated for 1,3,5,10 and 15 min respectively,then,fixed by 4% paraformaldehyde.The cells in second group were moved from hypertonic culture medium to isotonic culture medium and incubated for 1,3,5 and 10 min respectively,they were fixed too.These cells were stained by anti-Cx43 or anti-Cx32 immunofluorescent method and observed under confocal microscope. Results In first group,the expression of Cx43 increased then incubation for 1 min,at 3 min peaked,then decreased,at 15 min backed to normal level.The expression of Cx32 peaked at 5 min,later decreased,at 15 min backed to normal level.In second group,the expression of Cx43 also increased after incubation for 1 min,at 3 min peaked,then decreased,at 10 min backed to normal level.Cx32 expression peaked at 5 min,then decreased,at 10 min backed to normal level.The peak of Cx43 expression was early than that of Cx32.Conclusion Cx43 and Cx32 might be involved in osmoregulation.