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Methods Mol Biol. 2014;1124:121-58. doi: 10.1007/978-1-62703-845-4_9.

Analysis of electrophysiological properties and responses of neutrophils.

Author information

1
Department of Molecular Biophysics and Physiology, Rush University Medical Center, Chicago, IL, USA.

Abstract

The past decade has seen increasing use of the patch-clamp technique on neutrophils and eosinophils. The main goal of these electrophysiological studies has been to elucidate the mechanisms underlying the phagocyte respiratory burst. NADPH oxidase activity, which defines the respiratory burst in granulocytes, is electrogenic because electrons from NADPH are transported across the cell membrane, where they reduce oxygen to form superoxide anion (O2 (-)). This passage of electrons comprises an electrical current that would rapidly depolarize the membrane if the charge movement were not balanced by proton efflux. The patch-clamp technique enables simultaneous recording of NADPH oxidase-generated electron current and H(+) flux through the closely related H(+) channel. Increasing evidence suggests that other ion channels may play crucial roles in degranulation, phagocytosis, and chemotaxis, highlighting the importance of electrophysiological studies to advance knowledge of granulocyte function. Several configurations of the patch-clamp technique exist. Each has advantages and limitations that are discussed here. Meaningful measurements of ion channels cannot be achieved without an understanding of their fundamental properties. We describe the types of measurements that are necessary to characterize a particular ion channel.

PMID:
24504950
PMCID:
PMC3981097
DOI:
10.1007/978-1-62703-845-4_9
[Indexed for MEDLINE]
Free PMC Article

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