RECONSTITUTION OF A NEURONAL TRANSMITTER RELEASE SYSTEM

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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.