Regulated secretion and cytoskeleton in synapses.

1.神经递质转运蛋白寡肽的合成和抗肽抗体的产生存在于突触前膜中的神经递质转运蛋白作用于重新摄取释放到突触间隙中的神经递质，从而终止突触后反应。许多神经递质转运蛋白的cDNA已被克隆。本研究检测了多巴胺转运体、5-羟色胺转运体和谷氨酸转运体。设计了它们的氨基酸序列的特定部分，并化学合成了寡肽。以这些寡肽为抗原，采用体外免疫的方法制备抗肽抗体。2.多巴胺转运蛋白在PC12细胞分化过程中的表达。用NGF处理后的PC12细胞(h克隆)中检测多巴胺转运蛋白(DAT)的表达和亚细胞定位。在NGF作用下，DAT的表达首先在高尔基体区明显增强，然后在细胞表面表达。然后，表面积向外延伸，形成轴突。神经突起的尖端，即生长锥体，被强烈地标记着抗体。胞体胞浆和突起内可见许多抗体阳性反应的小斑点。

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.