Ishii K, Oda Y, Ichikawa T, Deguchi T
Department of Molecular Neurobiology, Tokyo Metropolitan Institute for Neurosciences, Japan.
Brain Res Mol Brain Res. 1990 Feb;7(2):151-9. doi: 10.1016/0169-328x(90)90092-r.
Complementary DNA clones containing the entire coding region of choline acetyltransferase (ChAT) were isolated from the spinal cords of rat and mouse. The cDNAs of rat and mouse coded for 640 and 641 amino acids, respectively, and showed 95% mutual homology and 80% homology with the cDNA of porcine ChAT. Northern blot analysis revealed a single band of 4.4 kb in the spinal cord and brain in each species. Introduction of the cDNAs into Chinese hamster ovary cells and neuron-derived cell lines, N1E115 and NG108-15, expressed a high ChAT activity, which was inhibited by a specific ChAT inhibitor. In situ hybridization using the rat cRNA probe revealed specific labeling of the motoneurons in the spinal cord and neurons in various forebrain nuclei of the rat where the existence of cholinergic neurons has been demonstrated immunohistochemically.
从大鼠和小鼠的脊髓中分离出了包含胆碱乙酰转移酶（ChAT）完整编码区的互补DNA克隆。大鼠和小鼠的cDNA分别编码640和641个氨基酸，彼此间具有95%的同源性，与猪ChAT的cDNA具有80%的同源性。Northern印迹分析显示，每个物种的脊髓和脑中均有一条4.4 kb的单带。将这些cDNA导入中国仓鼠卵巢细胞以及神经元衍生细胞系N1E115和NG108-15后，表达出了高ChAT活性，该活性可被一种特异性ChAT抑制剂所抑制。使用大鼠cRNA探针进行的原位杂交显示，在大鼠脊髓中的运动神经元以及前脑各个核团中的神经元出现了特异性标记，免疫组织化学已证实这些部位存在胆碱能神经元。
