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学科主题临床医学
Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation
Estacion, M.1,2,3; Choi, J. S.1,2,3; Eastman, E. M.1,2,3; Lin, Z.4; Li, Y.4; Tyrrell, L.1,2,3; Yang, Y.4; Dib-Hajj, S. D.1,2,3; Waxman, S. G.1,2,3
刊名JOURNAL OF PHYSIOLOGY-LONDON
2010-06-01
DOI10.1113/jphysiol.2009.186114
588期:11页:1915-1927
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Neurosciences ; Physiology
研究领域[WOS]Neurosciences & Neurology ; Physiology
关键词[WOS]HIGH-THROUGHPUT ELECTROPHYSIOLOGY ; ALPHA-SUBUNIT ; NA(V)1.7 CHANNEL ; SENSORY NEURONS ; VOLTAGE-CLAMP ; DOMAIN-IV ; PAIN ; ERYTHERMALGIA ; INACTIVATION ; MODULATORS
英文摘要

Ion channel missense mutations cause disorders of excitability by changing channel biophysical properties. As an increasing number of new naturally occurring mutations have been identified, and the number of other mutations produced by molecular approaches such as in situ mutagenesis has increased, the need for functional analysis by patch-clamp has become rate limiting. Here we compare a patch-clamp robot using planar-chip technology with human patch-clamp in a functional assessment of a previously undescribed Na(v)1.7 sodium channel mutation, S211P, which causes erythromelalgia. This robotic patch-clamp device can increase throughput (the number of cells analysed per day) by 3- to 10-fold. Both modes of analysis show that the mutation hyperpolarizes activation voltage dependence (-8 mV by manual profiling, -11 mV by robotic profiling), alters steady-state fast inactivation so that it requires an additional Boltzmann function for a second fraction of total current (similar to 20% manual, similar to 40% robotic), and enhances slow inactivation (hyperpolarizing shift -15 mV by human, -13 mV robotic). Manual patch-clamping demonstrated slower deactivation and enhanced (similar to 2-fold) ramp response for the mutant channel while robotic recording did not, possibly due to increased temperature and reduced signal-to-noise ratio on the robotic platform. If robotic profiling is used to screen ion channel mutations, we recommend that each measurement or protocol be validated by initial comparison to manual recording. With this caveat, we suggest that, if results are interpreted cautiously, robotic patch-clamp can be used with supervision and subsequent confirmation from human physiologists to facilitate the initial profiling of a variety of electrophysiological parameters of ion channel mutations.

语种英语
WOS记录号WOS:000278195900020
项目编号NCET-06-0015 ; 111039
资助机构Medical Research Service and Rehabilitation Research Service ; Department of Veterans Affairs ; National Multiple Sclerosis Society ; Erythromelalgia Association ; Program for New Century Excellent Talents in University ; Fok Ying Tong Education Foundation
引用统计
被引频次:23[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/58359
专题北京大学第一临床医学院_皮肤性病科
作者单位1.Yale Univ, Sch Med, Dept Neurol, New Haven, CT 06510 USA
2.Vet Affairs Connecticut Healthcare Syst, Rehabil Res Ctr, West Haven, CT 06516 USA
3.Yale Univ, Sch Med, Ctr Neurosci & Regenerat Res, New Haven, CT 06510 USA
4.Peking Univ, Hosp 1, Dept Dermatol, Beijing 100034, Peoples R China
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Estacion, M.,Choi, J. S.,Eastman, E. M.,et al. Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation[J]. JOURNAL OF PHYSIOLOGY-LONDON,2010,588(11):1915-1927.
APA Estacion, M..,Choi, J. S..,Eastman, E. M..,Lin, Z..,Li, Y..,...&Waxman, S. G..(2010).Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation.JOURNAL OF PHYSIOLOGY-LONDON,588(11),1915-1927.
MLA Estacion, M.,et al."Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation".JOURNAL OF PHYSIOLOGY-LONDON 588.11(2010):1915-1927.
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