Rieske E, Schubert P, Kreutzberg G W
Brain Res. 1975 Feb 14;84(3):365-82. doi: 10.1016/0006-8993(75)90759-3.
Intracellular application of tritiated precursors by means of microiontophoresis was performed on nerve cells in isolated segmental ganglia of the leech ventral nerve cord. Incorporation as well as intra- and interneuronal transport were studied by autoradiography after injection of fucose, glucosamine, glycine, leucine, orotic acid and uridine. With several minutes of intraneuronal injection the precursors were incorporated into macromolecules. Depending upon the tracer used, the radioactive material was distributed in a specific pattern over the cell somata and then released into the nerve processes. After application of orotic acid and uridine a transport of radioactive material, presumably RNA, could be observed in the processes of the injected neurons at a distance of about 200-500 mum. Fucose and glucosamine injection resulted in the most extended labeling of the nerve cell processes, indicating a transport rate of about 11 mm/day. When the radiochemicals were injected into one of the two electrically coupled giant nerve cells -- the so-called Retzius cells (Rc) -- a specific labeling not only of the injected Rc but also of the coupled but not injected Rc was found. Injection of protein or glycoprotein precursors into one Rc produced heavy labeling of both Rcs including their processes; a slight labeling of other ganglion compartments was only found after increasing the dosage of the amino acids glycine and leucine. With orotic acid and uridine this interneuronal transfer was confined to the electrically coupled Rc twin. Intracellular injection of one Rc with puromycin followed by injection of amino acids or fucose into the same Rc or into the coupled Rc resulted in an inhibition of precursor incorporation within the puromycin-injected Rc and an exclusive labeling of the coupled Rc, thus indicating that the precursors themselves were transferred. It is suggested that after microiontophoretic application an interneuronal transfer of relatively low molecular weight material takes place, probably across the low-resistance junction through which the Rcs are electrically coupled.
通过微离子电泳法将氚标记的前体物质注入水蛭腹神经索离体节段神经节的神经细胞内。在注入岩藻糖、葡糖胺、甘氨酸、亮氨酸、乳清酸和尿苷后,通过放射自显影研究其掺入以及神经元内和神经元间的转运情况。经过几分钟的神经元内注射,前体物质被掺入大分子中。根据所使用的示踪剂不同,放射性物质以特定模式分布在细胞体上,然后释放到神经突起中。注入乳清酸和尿苷后,在注射神经元的突起中,距注射点约200 - 500微米处可观察到放射性物质(可能是RNA)的转运。注入岩藻糖和葡糖胺导致神经细胞突起的标记范围最广,表明转运速率约为11毫米/天。当将放射化学物质注入两个电耦合的巨神经细胞之一——即所谓的雷丘斯细胞(Rc)时,不仅发现注入的Rc有特异性标记,而且未注射但与之耦合的Rc也有特异性标记。向一个Rc中注入蛋白质或糖蛋白前体,会使两个Rc及其突起都产生强烈标记;仅在增加甘氨酸和亮氨酸氨基酸剂量后,才在其他神经节区域发现轻微标记。对于乳清酸和尿苷,这种神经元间的转移仅限于电耦合的Rc对。先向一个Rc内注射嘌呤霉素,然后再向同一个Rc或与之耦合的Rc中注射氨基酸或岩藻糖,结果导致嘌呤霉素注射的Rc内前体物质掺入受到抑制,并使耦合的Rc出现特异性标记,从而表明前体物质本身发生了转移。有人提出,微离子电泳注入后,分子量相对较低的物质可能通过Rc之间电耦合的低电阻连接进行神经元间转移。