Ambron R T, Schacher S, Rayport S G
J Neurosci. 1985 Nov;5(11):2866-73. doi: 10.1523/JNEUROSCI.05-11-02866.1985.
The cell body of R2, a giant cholinergic neuron of Aplysia californica, resides in the abdominal ganglion, whereas its synapses are on thousands of unicellular mucus glands located in the skin. Due to the great spatial separation between the site of macromolecular synthesis and the presynaptic terminals, rapid axonal transport can be used to segregate synaptic proteins from those to be used elsewhere in the cell. The proteins of R2 were labeled by incubating the abdominal ganglion in [35S]methionine for 5 hr in a chamber separated from the rest of the isolated central nervous system. After 50 hr, 28 radiolabeled proteins were reproducibly found by one- and two-dimensional polyacrylamide gel electrophoresis to be transported to the distal regions of peripheral nerves P6, P7, and P8 that innervate the parapodia and middle body wall. We are sure that R2 is the source of these proteins since radioautography of sections taken throughout the nervous system, complemented by cobalt tracings, showed that R2 is the only neuron in the abdominal ganglion with axons in these nerves. Nine of the 28 transported proteins are glycoproteins since they were also labeled after injecting R2's cell body with [3H]-L-fucose. There is evidence that the proteins and glycoproteins are destined for R2's presynaptic terminals. For example, in experiments in which the body wall and parapodium remained attached to the nerves, the proteins were transported to the skin region that contains the glands. Moreover, analyses of the distribution of the rapidly transported proteins by qualitative radioautography and by extrusion of axoplasm indicated that none are constituents of the axolemma.(ABSTRACT TRUNCATED AT 250 WORDS)
加州海兔的巨大胆碱能神经元R2的细胞体位于腹神经节,而其突触则位于皮肤中数千个单细胞黏液腺上。由于大分子合成位点与突触前终末之间存在巨大的空间间隔,快速轴突运输可用于将突触蛋白与细胞其他部位使用的蛋白区分开来。通过在与分离的中枢神经系统其他部分隔开的腔室中,将腹神经节在[35S]甲硫氨酸中孵育5小时来标记R2的蛋白质。50小时后,通过一维及二维聚丙烯酰胺凝胶电泳可重复性地发现,有28种放射性标记蛋白被运输到支配副足和身体中壁的外周神经P6、P7和P8的远端区域。我们确定R2是这些蛋白质的来源,因为对整个神经系统切片进行放射自显影,并辅以钴追踪显示,R2是腹神经节中唯一轴突位于这些神经中的神经元。28种被运输的蛋白质中有9种是糖蛋白,因为在用[3H]-L-岩藻糖注射R2的细胞体后它们也被标记了。有证据表明这些蛋白质和糖蛋白是运往R2的突触前终末的。例如,在体壁和副足与神经相连的实验中,蛋白质被运输到含有腺体的皮肤区域。此外,通过定性放射自显影和轴浆挤出对快速运输蛋白质的分布进行分析表明,它们都不是轴膜的成分。(摘要截取自250词)