原代培养大鼠皮层神经细胞膜NMDAr蛋白冰冻蚀刻电镜研究

Freeze-etching electron microscopic study on immunocomplex of NMDA receptor protein on membrane of primary cultured rat cortical neurons

目的研究N-甲基-D-天冬氨酸受体（NMDAr）蛋白免疫金颗粒复合物在神经元膜超微结构及定位。方法应用冰冻蚀刻电镜技术对神经元膜NMDAr蛋白复合物进行研究。结果沿神经细胞膜面可见30—50nm免疫金复合物散在或密集分布，大小均匀。细胞核上密集均匀分布膜内微粒，4～16nm，与胶体金颗粒（CGP）区别较大。结论膜蛋白免疫金复合物沿膜面散在分布，大小均匀。冰冻蚀刻复型-透射电镜技术具有NMDAr蛋白免疫胶体金颗粒免疫复合物原位纳米尺度定位能力。

Objective To explore the localization and pathological characteristics of NMDA receptur protein immunocomplex on neuron membrane. Methods By using primary cuhured rat cortical neurons, the NMDA receptor protein immunocomplex on neuron membrane was observed by freeze - etching electron microscopic （FEEM） examination. Results Imrnunocomplex particles appeared on the corresponding structures of membrane surface were unevenly sized and distributed. The conformation of IgG - SPA - gold immunocomplex scanned through FEEM showed that its appearance is global or cosh in shape and its average diameter is 30 - 50 nm. The average diameter of particles on nucleolus is 4 - 16 nm. Conclusion The average diameter of immunocomplex is scattering distribution, uniforming size. The localization and shape of NMDAr protein immunocomplex on membrane surface of neurons can be determined by FEEM in nanometer scale.