High-Throughput Automated Patch Clamp System

摘要 高通量的电生理学技术正在彻底改变传统的人工膜片钳研究方法 渠道功能和加快药物发现的步伐。该提案要求购买一个 SyncroPatch 384 i工作站能够同时对384个细胞进行膜片钳测量。这么高- 吞吐量自动电生理系统将使休斯顿-加尔维斯顿地区的研究人员能够 进行大规模离子通道功能分析并筛选通道活性调节剂。初始组的 用户将分享这个仪器系统的不同项目，由25个单独的NIH赠款资助。两个合作 用户实验室将应用SyncroPatch通过筛选来识别和表征新的通道视紫红质变体 从序列数据库的候选人。视紫红质是真核生物的光门控离子通道， 神经科学研究人员广泛使用微生物来控制神经元和肌细胞的兴奋性， 动物模型（光遗传学），并在临床试验中作为光遗传基因疗法，以恢复盲人的视力。的 SyncroPatch将能够高通量选择已知通道视紫红质的突变群体， 优化和扩大其效用。离子通道病是无数损害大脑的人类疾病的原因， 心脏、免疫系统和其他功能。本申请中的十个建议用户研究离子通道， 分子和细胞的方法，以及动物模型，以研究在人类的通道的作用， 疾病和基本的生物机制，这些病理。因此，他们的实验室将应用 要求工作站：（1）筛选电压门控Na+通道、HCN通道、TRP的新型调节剂 通道、BK通道、耳蜗阴离子转运蛋白普雷斯廷（SLC 26 A5）和nAChR通道;（2）绘制耳蜗内的 离子型NMDA受体的构象景观;（3）确定Ca 2+的结构-功能决定因素 通道;（4）筛选胞吐作用的调节剂;和（5）分析突变和翻译后修饰， TRP、ASIC和TPC通道。SyncroPatch 384 i的收购将满足 Houston-Galveston生物医学研究中的自动化高通量膜片钳电生理学 社区，并将使我们能够以前所未有的速度询问离子通道功能，加速 新的光遗传学工具和新疗法的发现，从而拓宽了生物医学研究的视野 和药物开发。

ABSTRACT High-throughput electrophysiology is revolutionizing traditional approaches of manual patch clamp to study ion channel function and accelerating the pace of drug discovery. This proposal requests the purchase of a SyncroPatch 384i workstation capable of simultaneous patch clamp measurements on 384 cells. This high- throughput automated electrophysiology system will enable researchers in the Houston-Galveston area to conduct large-scale analysis of ion channel function and screen channel activity modulators. The initial group of users will share this instrument system for diverse projects, funded by 25 individual NIH grants. Two collaborating user labs will apply the SyncroPatch to identify and characterize new channelrhodopsin variants by screening candidates from sequence databases. Channelrhodopsins are light-gated ion channels from eukaryotic microorganisms widely used by neuroscience researchers to control excitability of neurons and myocytes in animal models (optogenetics), and as optogenetic gene therapy in clinical trials to restore vision to the blind. The SyncroPatch will enable high-throughput selection of mutated populations of known channelrhodopsins to optimize and expand their utility. Ion channelopathies are the cause of myriad human diseases that impair brain, cardiac, the immune system, and other functions. Ten proposed users in this application study ion channels with molecular and cellular methods, as well as animal models, to investigate the role of the channels in human diseases and basic biological mechanisms underlying these pathologies. Accordingly, their labs will apply the requested workstation to: (1) screen for novel modulators of voltage-gated Na+ channels, HCN channels, TRP channels, BK channels, the cochlear anion transporter prestin (SLC26A5) and nAChR channels; (2) map the conformational landscape of ionotropic NMDA receptors; (3) identify structure-function determinants of Ca2+ channels; (4) screen modulators of exocytosis; and (5) analyze mutations and post-translational modifications of TRP, ASIC and TPC channels. The acquisition of the SyncroPatch 384i will answer the urgent need for automated high-throughput patch-clamp electrophysiology in the Houston-Galveston biomedical research community and will enable us to interrogate ion channel function at an unprecedented pace, accelerate discoveries of new optogenetic tools and new therapeutics, and thus broaden the horizon of biomedical research and drug development.