Synuclein Function in the Synaptic Vesicle Cycle

几个独立的证据已经将富含神经末梢的突触核蛋白家族与帕金森病联系起来。遗传学研究已经将α-突触核蛋白基因的突变与几个家庭的疾病联系起来。突触核蛋白是路易体的主要成分，路易体是帕金森病的标志性组织病理学。突触核蛋白的过度表达是路易体的主要成分，路易体是帕金森病的标志性组织病理学。突触核蛋白在小鼠中的过度表达导致路易体样神经病理学和运动缺陷。虽然有大量的信息的遗传和组织学特征的突触核蛋白，很少的信息存在的蛋白质在正常功能的神经末梢的功能。我们建议研究这些蛋白质在神经末梢中的功能，使用经典的神经生理学实验在一个孤立的神经末梢中使用经典的神经生理学实验在一个孤立的神经末梢。将修改突触核蛋白功能的药物注射到原始脊椎动物（七鳃鳗）的巨大前末端。将采用电生理学和超微结构方法分析终末前注射几种突触核蛋白影响剂的影响。受感染的病原体包括：七鳃鳗突触核蛋白的全长和片段，含有帕金森病相关突变（A53 T和A30 P）的重组人α-突触核蛋白。模拟酪蛋白激酶I和src激酶磷酸化（S129 E/S87 E）人α-突触核蛋白和针对七鳃鳗突触核蛋白的抗体。使用成对的细胞内记录之间的前和突触后的元素，我们将检查注射对由内oxonal刺激产生的ESPS的影响。突触囊泡释放动力学的许多参数将被检查。此外，将在电子显微镜下检查接受药物注射后接受不同水平刺激的巨大突触前元件。注射对突触囊泡周期的影响可以通过神经末梢形态的特征性变化来确定。建立突触核蛋白在正常神经递质释放中的功能，将有助于阐明帕金森病中蛋白质的功能和功能障碍。

Several independent lines of evidence have linked the nerve terminal- enriched synuclein family of proteins to Parkinson's disease. Genetic studies have linked mutations in the alpha-synuclein gene to the disease in several families. Synucleins are a major constituent of Lewy bodies, a hallmark histopathology of Parkinson's disease. Over-expression of synucleins are a major constituent of Lewy bodies, a hallmark histopathology of Parkinson's disease. Over-expression of synucleins in mice causes Lewy body-like neuropathology and motor deficits. While there is a wealth of information on the genetic and histological features of synucleins, very little information exists on the function of the proteins in normally functioning nerve terminals. We propose to study the function of these proteins in nerve terminals using classic neurophysiologic experiments in an isolated nerve terminals using classic neurophysiologic experiments in an isolated nerve terminal. Pre-synaptic injection of agents that will modify synuclein function will be made into the giant pre-terminal of a primitive vertebrate (lamprey). The effects of pre-terminal injections of several synuclein affecting agents will be analyzed with electrophysiologic and ultrastructural methods. Inj3ected agents will include: full length and fragments of lamprey synuclein, recombinant human alpha-synuclein containing Parkinson's disease associated mutations (A53T and A30P). Mock casein kinase I and src kinase phosphorylated (S129E/S87E) human alpha-synuclein and antibodies against lamprey synuclein. Using paired intracellular recordings between the pre- and post-synaptic elements we will examine the effects of injections on the ESPS generated by intraoxonal stimulation. Many parameters of synaptic vesicle release kinetics will be examined. In addition, giant pre-synaptic elements receiving injections of agents followed by varying levels of stimulation will be examined under the electron microscope. Effects of injections on the synaptic vesicle cycle can be established by characteristic changes in nerve terminal morphology. Establishing the function of the synucleins in normal neurotransmitter release will shed light on the function and dysfunction of the protein in Parkinson's disease.