论文部分内容阅读
目的探讨磷酸化突触素I在微波辐射所致囊泡循环和氨基酸递质释放改变中的作用。方法采用30 m W/cm2微波辐射原代海马神经元,免疫荧光染色观察辐射后1、6、12 h,1、2、3 d神经元Ser-62/67及Ser-553位点突触素I磷酸化表达改变;高效液相色谱检测辐射后6 h海马神经元氨基酸递质释放量变化;time-lapse观察辐射后6 h神经元突触囊泡循环改变;给予U0126和roscovitine进行干预,观察上述指标改变。结果 30 m W/cm2微波辐射后海马磷酸化突触素I Ser-62/67、Ser-553及磷酸化细胞外调节激酶(p-ERK)、细胞周期素依赖性激酶(Cdk5)表达明显减少,γ-氨基丁酸(GABA)释放明显减少,海马神经元囊泡循环速度减慢;辐射并给予U0126后,神经元p-ERK和Ser-62/67位点突触素I磷酸化表达明显减弱,囊泡循环速度减慢;辐射并给予roscovitine后,神经元Cdk5表达减少,Ser-553位点突触素I磷酸化表达上调,GABA释放明显增加,囊泡循环速度明显减慢。结论 30 m W/cm2微波辐射通过下调海马神经元ERK活性抑制突触素I Ser-62/67位点磷酸化而使囊泡循环受抑制,囊泡锚定障碍;通过下调Cdk5促进突触素I Ser-553位点磷酸化而利于含GABA的囊泡锚定于突触前膜,但抑制囊泡循环。
Objective To investigate the role of phosphorylated synaptophysin I in vesicle cycle and amino acid transmitter release induced by microwave irradiation. Methods Primary cultured hippocampal neurons were exposed to 30 mW / cm2 microwave radiation. The expression of synaptophysin at the Ser-62/67 and Ser-553 sites of neurons at 1, 6, 12 h, I phosphorylation expression changes; high-performance liquid chromatography 6 h after irradiation the amino acid neurotransmitter release changes; time-lapse observed 6 h after irradiation neuronal synaptic vesicle cycle changes; give U0126 and roscovitine intervention, observation The above indicators change. Results The phosphorylated synaptophysin I Ser-62/67, Ser-553 and phosphorylated extracellular regulated kinase (p-ERK) and cyclin dependent kinase (Cdk5) in hippocampus after 30 mW / cm2 microwave irradiation were significantly decreased , the release of γ-aminobutyric acid (GABA) was significantly reduced, and the circulation of vesicles in hippocampal neurons was slowed down. After U0126 irradiation, the expression of synaptophysin I in neurons p-ERK and Ser-62/67 was significantly increased Decreased, and the vesicle circulation slowed down. After irradiation and administration of roscovitine, the expression of Cdk5 in neurons decreased, the phosphorylation of Ser-553 phosphorylation of synaptophysin I was increased, the release of GABA was significantly increased, and the rate of vesicle cycle slowed down significantly. Conclusion Microwave irradiation of 30 mW / cm2 can inhibit the vesicle circulation and inhibit the vesicle disruption by inhibiting the phosphorylation of the phosphorylated synaptophysin I Ser-62/67 by downregulating the ERK activity of hippocampal neurons. Phosphorylation of the I Ser-553 site favors anchoring of GABA-containing vesicles to the presynaptic membrane, but inhibits vesicle cycling.