Regulated secretion and cytoskeleton in synapses.

点击翻译按钮获取中文摘要

1.Synthesis of oligopeptides of the neurotransmitter transporter and production of the anti-peptide antibodiesThe neurotransmitter transporters presenting in the presynaptic membrane work to reuptake the neurotransmitter released into the synaptic cleft and thereby to terminate the postsynaptic response. cDNA of many neurotransmitter transporters have been cloned. In the present study dopamine transporter, serotonin transporter and glutamate transporter were examined. The specific parts of their amino acid sequences were designed and oligopeptides were chemically synthesized. Then, the antipeptide antibodies were produced with in vitro immunization method using these oligopeptides as antigens. These antibodies were characterized the specificity to the original transporter proteins by Western blotting.2.Expression of dopamine transporter in the PC12 cells during differentiation.The expression and subcellular localization of dopamine transporter(DAT)was examined in the PC12 cells (h-clone) treated with NGF.DAT was immunohistochemically detected on the surface of the PC12 cells. When the cells were treated with NGF,the expression of DAT was significantly emphasized first in the Golgi area and then on the surface of the cell. The surface area, then, extended outwards forming neurites. The tips of the neurites, growth cones, were strongly labeled with the antibody. Many small spots positively responsing to the antibody were detected in the cytoplasm of the cell body and in the neurites.