One-channel cell-attached patch-clamp recording

Bruce A. Maki, Kirstie A. Cummings, Meaghan A. Paganelli, Swetha E. Murthy, Gabriela K. Popescu

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate depends on properties intrinsic to each channel protein as well as the mechanism by which it is generated and controlled and represents an important area of current research. Information about the operational dynamics of ion channel proteins can be obtained by observing long stretches of current produced by a single molecule. Described here is a protocol for obtaining one-channel cell-attached patch-clamp current recordings for a ligand gated ion channel, the NMDA receptor, expressed heterologously in HEK293 cells or natively in cortical neurons. Also provided are instructions on how to adapt the method to other ion channels of interest by presenting the example of the mechano-sensitive channel PIEZO1. This method can provide data regarding the channel's conductance properties and the temporal sequence of open-closed conformations that make up the channel's activation mechanism, thus helping to understand their functions in health and disease.

Original languageEnglish (US)
JournalJournal of Visualized Experiments
Issue number88
DOIs
StatePublished - Jun 9 2014
Externally publishedYes

Keywords

  • Biophysics
  • Electrophysiology
  • Gating
  • Ion channels
  • Issue 88
  • Kinetic analysis
  • Neuroscience
  • NMDA receptors
  • Patch-clamp
  • Single-channel recording

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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