Tandai-Hiruma M, Mori-Okamoto J, Kotani M, Miura K, Takishima K, Nishida Y
Department of Physiology, National Defense Medical College, Tokorozawa, Saitama, Japan.
Neurochem Res. 2000 Aug;25(8):1095-106. doi: 10.1023/a:1007670012531.
Among several monoclonal antibodies obtained by immunizing Balb/c mice with cerebellar synaptic membrane fractions from E20 chick embryos, the antibody, named M35, suppressed Ca-spikes in immature cultured chick cerebellar neurons. M35 immunoprecipitated 43 kDa protein from a 125I-labeled embryonic crude cerebellar membrane fraction. Immunohistochemically, the M35 antigen was expressed most intensively in Purkinje cells, but its expression was limited to highly motile structures at developmental neuronal remodeling. Electrophysiologically, M35 facilitated current responses to AMPA and inhibited the responses to GABA in cultured cerebellar Purkinje neurons. The several peptides derived from the affinity-purified 43 kDa protein were found to have homologous amino acid sequences to non-muscle actins. These results suggest that the antigen recognized by M35 may play an essential role probably as membrane ion channels modulating synaptic functions in not only the development and growth but also the neuronal activity of chick cerebellar Purkinje cells.
在用E20鸡胚的小脑突触膜组分免疫Balb/c小鼠获得的几种单克隆抗体中,名为M35的抗体可抑制未成熟培养的鸡小脑神经元中的钙峰。M35从125I标记的胚胎小脑粗膜组分中免疫沉淀出43 kDa的蛋白质。免疫组织化学显示,M35抗原在浦肯野细胞中表达最为强烈,但其表达仅限于发育中神经元重塑时的高迁移结构。电生理学研究表明,M35可促进培养的小脑浦肯野神经元对AMPA的电流反应,并抑制对GABA的反应。从亲和纯化的43 kDa蛋白质衍生的几种肽被发现与非肌肉肌动蛋白具有同源氨基酸序列。这些结果表明,M35识别的抗原可能不仅在鸡小脑浦肯野细胞的发育、生长以及神经元活动中,而且在调节突触功能的膜离子通道方面发挥重要作用。