Unified patch clamp protocol for the characterization of Pannexin 1 channels in isolated cells and acute brain slices

Christina Gründken, Julian Hanske, Stephanie Wengel, Wiebke Reuter, Amr Abdulazim, Valery I. Shestopalov, Rolf Dermietzel, Georg Zoidl, Nora Prochnow

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

In the central nervous system, Pannexin 1 (Panx1) channels are implicated in a variety of physiological and pathological conditions. One of the prerequisites to enlighten the role of Panx1 is the development and standardization of reliable methods. Here, we address the applicability of voltage clamp protocols to identify Panx1 channel mediated currents in neurons of acutely dissected brain slices. We improved an established protocol and report on a modified paradigm that robustly evokes Panx1 channel currents. Crucial advances are the use of physiologic ion gradient conditions and a preconditioning step of depolarizing membrane potential ramps of long duration. This new paradigm provides significant impact on membrane current generation at hypo- and depolarized holding potential steps post voltage ramp preconditioning in heterologous expression systems and primary hippocampal CA1 neurons of mouse brain slices in vitro. Finally, we demonstrate that under these conditions the analysis of tail currents elicited by repolarization of the cells from preconditioning holding potential depolarization permits an independent method to isolate Panx1 mediated channel activity. In summary, this study provides a comprehensive methodological improvement in the biophysical analysis of Panx1 channels with a particular focus on investigations under physiological conditions in complex tissues.

Original languageEnglish (US)
Pages (from-to)15-25
Number of pages11
JournalJournal of Neuroscience Methods
Volume199
Issue number1
DOIs
StatePublished - Jul 15 2011

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Keywords

  • Hippocampus
  • N2a cells
  • Neuron
  • Pannexin 1
  • Tail current

ASJC Scopus subject areas

  • Neuroscience(all)

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