Insights Into The Mechanisms Of Sema3F-induced Growth Cone Collapse In Primary Rat Hippocampal Neurons

Objective To investigate the changes of VEGF concentration over time and the effect of Sema3 F on the concentration of VEGF in cultured primary hippocampal system.Methods Hippocampal neurons were dissociated from Wistar rats within postnatal 24 hours and cultured in primary system.The concentration of VEGF in the culture medium was measured by using Enzyme-Linked Immunosorbent Assay in the first day,third day,fifth day,seventh day,ninth day,respectively.Sema3 F with appropriate final concentration was added to the culture medium of hippocampal neurons in the third day,then VEGF concentration were measured immediately,5 minutes later,15 minutes later,30 minutes later,respectively.Results(1)The culture system of hippocampal neurons in rats was successfully established in vitro.(2)The concentration of VEGF on the 1st day,3rd day,5th day,7th day and 9th day in the experimental group was higher than that in the control group at the corresponding time respectively;(3)Primary rat hippocampal neurons were cultured on the 3rd day,the results showed that the concentration of VEGF increased gradually at 4 different time points in the control group,then were treated by Sema3 F with the most appropriate concentration in the experimental group,the concentration of VEGF was the highest 5 minutes later,and then decreased gradually as time goes on.Conclusions(1)VEGF is autocrine in primary hippocampal neuronal culture system of rats.(2)Sema3F can induce the change of VEGF concentration in primary hippocampal neuronal supernatant of rats.