G PROTEIN MODULATION OF N-TYPE CALCIUM CHANNELS

描述：(申请人描述) 这项研究建议的长期目标是了解 与N-型钙通道调控相关的分子事件 Gbetagamma亚基。需要检验的主要假设是，亚基 Gbetagamma二聚体的组成，即Gbeta和Gbeta的特定组合 GGamma对电压依赖的N型钙通道具有特异性 调制。因此，特定的目标是1)识别哪种G蛋白 β亚基对大鼠电压依赖性N型钙通道的抑制作用 交感神经元；2)确定哪种G蛋白结合 赋予电压依赖的N型钙通道调节的特异性；以及3) 鉴定天然表达的G蛋白β亚基和伽马亚基 用Western blotting方法对大鼠交感神经元进行免疫组织化学染色。通过 利用电生理、分子和生化方法， 将描述钙通道调节中的Gbetagamma组成 更好地理解。 N型钙通道的电压依赖性调制已在 中枢神经系统和外周神经系统的神经元种类繁多。 尽管大多数研究都是在神经元胞体上进行的，但有证据表明 神经末梢的直接研究支持这种形式的神经末梢 调制在突触前突触机制中起重要作用 变速箱。因此，有很大的兴趣进一步发展我们的 对参与N型钙通道的分子事件的理解 调制。

DESCRIPTION: (Applicant's Description) The long-term goal of this research proposal is to gain an understanding of the molecular events associated with the modulation of N-type Ca2+ channels by Gbetagamma subunits. The main hypothesis to be tested is that the subunit composition of the Gbetagamma dimer, i.e., specific combinations of Gbeta and Ggamma confer specificity in regard to voltage-dependent N-type Ca2+ channel modulation. Accordingly, the specific aims are 1) to identify which G protein beta subunit produces voltage-dependent N-type Ca2+ channel inhibition in rat sympathetic neurons; 2) to determined which G protein betagamma combinations confer specificity for voltage-dependent N-type Ca2+ channel modulation; and 3) to identify the G protein beta and gamma subunits that are natively expressed in rat sympathetic neurons by employing the Western blotting method. By employing electrophysiological, molecular, and biochemical methods, the role of the Gbetagamma composition in modulation of Ca2+ channels will be delineated and better understood. Voltage-dependent modulation of N-type Ca2+ channels has been demonstrated in a wide variety of neurons from both the central and peripheral nervous system. Although most studies are performed on the neuron soma, there is evidence from direct studies of nerve terminals to support the idea that this form of modulation plays an important role in presynaptic mechanisms of synatic transmission. Thus there is substantial interest in furthering our understanding of the molecular events which contribute to N-type Ca2+ channel modulation.