Location and Function of VGLUT3

描述（由申请人提供）：谷氨酸是哺乳动物脑中的主要兴奋性神经递质，并被包装到分泌囊泡中，通过囊泡转运蛋白家族（VGLUT 1 -3）进行胞吐释放。令人惊讶的是，VGLUT 3在神经元的离散群体中表达，这些神经元以释放其他经典递质而闻名：多巴胺，血清素GABA和ACh。我的目标是确定VGLUT 3在单胺和胆碱能神经元中的精确细胞和亚细胞位置，特别关注载体与突触释放位点和含有单胺或ACh的囊泡的关系。第二个目的是评估VGLUT 3在谷氨酸释放中的作用。为了实现这一点，我将检查VGLUT 3的位置在小鼠的大脑中，表达荧光报告，仅在单胺神经元的光学和电子显微镜，以及生产和检查VGLUT 3基因敲除小鼠表达的荧光报告下的VGLUT 3启动子的控制。这些小鼠将用于免疫染色和电生理记录。这些发现将有助于我们更好地理解VGLUT 3在单胺神经元中的生理作用，并探索VGLUT 3表达对这些信号通路的影响，并最终影响受单胺神经元影响的行为，如运动控制，情感，奖励和药物成瘾。

DESCRIPTION (provided by applicant): Glutamate is the major excitatory neurotransmitter in the mammalian brain, and is packaged into secretory vesicles for exocytotic release by a family of vesicular transporters (VGLUT1-3). Surprisingly, VGLUT3 is expressed in discreet populations of neurons better known for their release of other classical transmitters: dopamine, serotonin GABA and ACh. I aim to determine the precise cellular and subcellular location of VGLUT3 in monoamine and cholinergic neurons, focusing in particular on the relationship of the carrier to sites of synaptic release and to vesicles that contain monoamines or ACh. A second aim is to evaluate the role of VGLUT3 in glutamate release. To achieve this, I will examine the location of VGLUT3 in the brains of mice that express a fluorescent reporter in only monoamine neurons by light and electron microscopy, as well as to produce and examine VGLUT3 knockout mice that express a fluorescent reporter under the control of the VGLUT3 promoter. These mice will be used for immunostaining and for electrophysiological recordings. These findings will help us to better understand the physiological role of VGLUT3 in monoamine neurons, as well as to explore the impact of VGLUT3 expression on these signaling pathways and ultimately on behaviors that are affected by monoamine neurons, such as motor control, affect, reward and drug addiction.