The Main Elements Of Quick Signal Transduction Pathway About Ciliary Neurotrophic Factor

In the previous study,we have discovered the quick protective effects of CNTF on hippocampal neuron injury in stress,but we still have little idea on its mechanism.To study the primary elements involving in quick pathway of CNTF,on the basis of NMDA-induced hippocampal neuron injury model,successive photography of vial cells,the whole-cell patch clamp and concentration measurement of intracellular free[Ca²⁺]were used to observe quick protection of CNTF on hippocampal neuron and the effects of PTPi-2(the blocker of JAK/STAT pathway) and anti-CNTFRαon the quick protection.The methods are as follows:First,the protective function of CNTF on neurons were observed by using the successive photography of vial cells.The effects of Anti-CNTFRαand PTPi-2 on the protective function were also studied.Second,the excitatory of hippocampal neurons were recorded by using the whole cell patch-clamp techniques.Third,the effects of CNTF,Anti-CNTFRαand PTPi-2 on NMDA-induced current were recorded by using the whole cell patch-clamp techniques.Forth,by using the concentration measurement of intracellular free[Ca²⁺],the effert of CNTF,Anti-CNTFRαand PTPi-2 on NMDA-induced[Ca²⁺]elevtion were recorded.Based on the results,the conclutions are as follows:First,the result of successive photography of vial cells suggests that CNTF can protect cells from stressful damage,and Anti-CNTFRαcan block this function,while PTPi-2 can not.Second,whole-cell patch clamp shows that there is no significant correlation between CNTF and hippocampal neuronal excitability,but CNTF can inhibit NMDA-induced potential.And anti-CNTFRαcan block this protective function,however PTPi-2 can not.Third,we can observe the inhibitory effects of CNTF on the elevation level of NMDA-induced intracellular free[Ca²⁺]by concentration measurement of intracellular free[Ca²⁺],and anti-CNTFRαcan block its function,but PTPi-2 can not.In summary,from the study that Anti-CNTFRαand PTPi-2 has impact on the quick roles of CNTF,we can infer that CNTFR,NMDAR and[Ca²⁺]is a part of CNTF quick pathway,while JAK/STAT pathway are not included.