免疫电镜技术在微波辐射致大鼠海马突触结构损伤研究中的应用

Application of immunoelectron microscopy on synaptic structure injury research of hippocampus in Wistar rats induced by microwave radiation

目的:通过免疫电镜技术的应用,探讨微波辐射致大鼠海马突触结构损伤的机制。方法:采用30mW/cm2微波辐射Wistar大鼠,通过电镜观察辐射后6h海马突触结构改变;采用不同实验条件摸索胶体金标记的免疫电镜染色方法,并检测磷酸化突触素I和囊泡氨基酸递质转运体的改变。结果:30mW/cm2微波辐射后大鼠海马突触前囊泡堆积,活性区延长、突触后致密物增加、突触曲率增加,GABA囊泡转运体和磷酸化突触素I(Ser-553)在海马堆积囊泡中表达增加。结论:胶体金标记的免疫电镜染色操作过程中,固定液中戊二醛浓度和处理时间对其结果影响至关重要;微波辐射可引起突触结构损伤,Ser-553位点突触素I磷酸化参与微波辐射所致GABA能囊泡的堆积。

Objective：To investigate the mechanism of synaptic structure injury after microwave radia-tion in hippocampus of rats by application of immunoelectron microscopy .Methods：Wistar rats were ex-posed to microwave radiation .Electron telescope was used to study the change of the synaptic structure at 6h after radiation .The different experiment conditions were used in colloidal gold immunoelectron micros-copy and to study the change of phosphorylated synapsin I and vesicular amino acids neurotransmitter transporters .Results：The average power density which was set at 30mW/cm2 ,0 .1% glutaraldehyde in fixative fluid and short time of sucrose dehydration were better for maintenance of ultraculture and anti-gens activity .It resulted in deposits of synapse vesicle ,elongation of active zone ,the increase of thickness of postsynaptic density and perforation of synapse ;the expressions of vesicular GABA transporter and phosphorylation of synapsin I at Ser-553 were increased in cumulated synapse vesicles .Conclusion：In the experiments of colloidal gold immunoelectron microscopy ,the concentration of glutaraldehyde in fixative fluid and the time of sucrose dehydration were important .Microwave radiation can induce the injure of synaptic structure and the phosphorylation of synapsin I at Ser-553 may participate in the deposits of GA-BA synapse vesicles .