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Equipment for brain slice patch-clamp electrophysiology

脑切片膜片钳电生理学设备

基本信息

批准号：
374732-2009
负责人：
Nashmi, Raad
金额：
$ 3.59万
依托单位：
University of Victoria
依托单位国家：
加拿大
项目类别：
Research Tools and Instruments - Category 1 (<$150,000)
财政年份：
2008
资助国家：
加拿大
起止时间：
2008-01-01 至 2009-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=412000
关键词：
Equipment brain slice patch clamp

项目摘要

Communication between neurons in the brain is governed by synaptic transmission, which is mediated by the presynaptic release of chemical neurotransmitters and their binding to cell surface receptors on the postsynaptic neuron. The research in our lab focuses on a class of ligand-gated ion channels called nicotinic acetylcholine receptors. Our long term goal is to examine the basic function of nicotinic receptors in modulating neurotransmission in the CNS. We have developed a novel approach to understanding nicotinic receptor mediated signaling in the brain. We have genetically engineered knock-in mice in which we fused a fluorescent protein to the alpha4 nicotinic receptor subunit. To our knowledge this is the first knock-in mouse in which an ion channel has been tagged with a fluorescent protein. Alpha4 beta2 nicotinic receptors constitute the major nicotinic receptor subtype in the CNS. For the first time, we have the ability to quantify accurately at submicron resolution the subcellular distribution of the receptor in neurons in the CNS. This important information will allow us to precisely target those neurons for electrophysiology to ascertain the function of alpha4 subunit containing nicotinic receptors. Furthermore, we have also generated knock-in mice with a gain-of-function mutation to the alpha4 nicotinic receptor gene. This mutation renders the receptor 50 fold more sensitive to activation by nicotine or acetylcholine and expresses four fold greater nicotinic responses thus facilitating the recording of alpha4 nicotinic receptors.

大脑中神经元之间的通信由突触传递控制，突触传递由化学神经递质的突触前释放及其与突触后神经元上的细胞表面受体的结合介导。我们实验室的研究集中在一类称为烟碱乙酰胆碱受体的配体门控离子通道上。我们的长期目标是研究烟碱受体在调节中枢神经系统神经传递中的基本功能。我们已经开发了一种新的方法来了解烟碱受体介导的信号在大脑中。我们有基因工程敲入小鼠，在其中我们融合了荧光蛋白和α 4烟碱受体亚基。据我们所知，这是第一个用荧光蛋白标记离子通道的基因敲入小鼠。α 4 β 2烟碱受体构成CNS中的主要烟碱受体亚型。这是第一次，我们有能力在亚微米分辨率下准确地量化CNS神经元中受体的亚细胞分布。这一重要信息将使我们能够精确地靶向这些神经元进行电生理学研究，以确定含有烟碱受体的α 4亚基的功能。此外，我们还产生了α 4烟碱受体基因功能获得性突变的敲入小鼠。这种突变使受体对尼古丁或乙酰胆碱激活的敏感性增加50倍，并表达4倍的烟碱反应，从而促进α 4烟碱受体的记录。