Linderman K M, Greif K F
Department of Biology, Bryn Mawr College, Pennsylvania 19010.
J Neurobiol. 1990 Mar;21(2):295-302. doi: 10.1002/neu.480210205.
The role of membrane depolarization in the regulation of expression of a neuron specific protein was evaluated by culturing superior cervical ganglia from neonatal rats in defined medium and manipulating neuronal activity by depolarizing agents. P65 is an integral membrane protein of synaptic vesicles and can be used as a marker for general neuronal maturation. P65 antigen levels were quantified by indirect radioimmunoassay, using monoclonal antibodies. The expression of p65 in ganglion explants increased by 40-100% when the cultures were treated with the depolarizing agents, veratridine or high potassium. The veratridine effect could be blocked by simultaneous treatment with the sodium channel blocker, tetrodotoxin (TTX). The rise in p65 was not evident until 36 h after depolarizing treatment had begun and reached peak levels after 48 h, with no further increases observed with sustained treatment. After removal of the depolarizing treatment, p65 levels returned to control values after 24 h. P65 joins a growing number of molecules whose expression is regulated by membrane depolarization.
通过在特定培养基中培养新生大鼠的颈上神经节，并使用去极化剂来操纵神经元活动，评估了膜去极化在调节神经元特异性蛋白表达中的作用。P65是突触小泡的一种整合膜蛋白，可作为一般神经元成熟的标志物。使用单克隆抗体通过间接放射免疫测定法定量P65抗原水平。当用去极化剂藜芦碱或高钾处理培养物时，神经节外植体中p65的表达增加了40%-100%。藜芦碱的作用可通过同时用钠通道阻滞剂河豚毒素（TTX）处理来阻断。直到去极化处理开始后36小时，p65的升高才明显，并在48小时后达到峰值水平，持续处理未观察到进一步增加。去除去极化处理后，24小时内p65水平恢复到对照值。P65加入了越来越多其表达受膜去极化调节的分子行列。
