Can robots patch-clamp as well as humans? Characterization of a novel sodium channel mutation.

The Journal of Physiology
M EstacionS G Waxman

Abstract

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 Nav1.7 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 (approximately 20% manual, approximately 40% robotic), and enhances slow inactivation (hyperpolarizing shift--15 mV by human,--13 mV robotic). Manual patch-clamping demonstrated slower deactivation and enhanced (approximately 2-fold) ramp response for the mutant channel ...Continue Reading

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Citations

Feb 10, 2010·The Journal of Physiology·Roman CreggJing Zhao
Apr 9, 2010·The Journal of Physiology·Dimitri M Kullmann, Stephen G Waxman
Jul 12, 2013·Annals of the New York Academy of Sciences·Suhasa B KodandaramaiahCraig R Forest
Oct 1, 2015·Orphanet Journal of Rare Diseases·Zhaoli TangHong Jiang
Feb 5, 2013·Biochemical and Biophysical Research Communications·Mark EstacionStephen G Waxman
May 14, 2016·Scientific Reports·Martin HamplAngelika Lampert
Oct 29, 2011·Vascular Medicine·Nedaa SkeikRandal C Richardson
Jan 28, 2012·Therapeutic Advances in Gastroenterology·Arthur Beyder, Gianrico Farrugia
May 23, 2014·Lab on a Chip·Brandon R BruhnMichael Mayer
Feb 27, 2017·BMC Systems Biology·Dimos KapetisUNKNOWN PROPANE Study Group
May 17, 2018·Brain and Neuroscience Advances·Luca A Annecchino, Simon R Schultz
Jul 1, 2020·Pflügers Archiv : European journal of physiology·Mark D Baker, Mohammed A Nassar
Sep 22, 2019·International Journal of Molecular Sciences·Cosmin I CiotuMichael J M Fischer
Sep 12, 2020·Biochimica Et Biophysica Acta. Molecular Basis of Disease·Laura SoléMichael M Tamkun
Apr 25, 2021·BMC Bioinformatics·Makros N XenakisHubert J Smeets

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