RECONSTITUTION OF A NEURONAL TRANSMITTER RELEASE SYSTEM

这个项目的目标是提高我们对发射机的理解 神经突触前事件的释放过程和调控手段 细胞 这种方法需要重建一种神经递质 释放系统使用非洲爪蟾卵母细胞。 这些卵母细胞可以是 显微注射各种材料，重要的是， 显示翻译外源提供的信使RNA。 重建 释放过程包括三个阶段。 首先，卵母细胞将 注射来自鱼雷神经元的信使RNA， 纳辛 这一步可能是至关重要的赋予卵母细胞的成分 需要支持递质的胞吐放电。 卵母细胞将 然后注射从电泳板分离的突触囊泡， Torpedo或Narcine组织。 最后，卵母细胞将被刺激 以确定是否诱发释放 乙酰胆碱（acetylcholine）可以被检测到。 成功的这项 重组工作将有助于详细审查 参与递质分泌的分子成分。 所得 观察可能为研究生物化学相关性开辟新的途径， 突触可塑性 此外，广泛的神经功能缺损（例如， 痴呆症、帕金森综合症、精神发育迟滞）可能会受到详细审查 以一种新的眼光。

The goal of this project is to improve our understanding of the transmitter release process and the means of regulating presynaptic events in nerve cells. The approach entails the reconstitution of a neuronal transmitter release system using the oocytes of Xenopus laevis. These oocytes can be microinjected with a wide range of materials, and importantly, have been shown to translate exogenously supplied messenger RNA. The reconstruction of the release process involves three phases. First, oocytes will be injected with messenger RNA from electromotor neurons of Torpedo or Narcine. This step may be crucial for endowing oocytes with components needed to support the exocytotic discharge of transmitter. The oocyte will then be injected with synaptic vesicles isolated from the electroplax tissue of Torpedo or Narcine. Finally, the oocytes will be stimulated electrically or chemically to determine whether evoked release of transmitter (acetylcholine) can be detected. Success of this reconstitution effort will facilitate a detailed examination of the molecular components involved in transmitter secretion. The resulting observations may open new avenues to the study of biochemical correlates of synaptic plasticity. Moreover, a wide range of neurological deficits (eg., dementia, Parkinsonism, mental retardation) might be amenable to scrutiny in a new light.