NDLI: Patch-Clamp Electrophysiology of Intracellular $Ca^{2+}$ Channels

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Mak, Don-On Daniel Vais, Horia Cheung, King-Ho Foskett, J. Kevin
Description	Country affiliation: United States Author Affiliation: Mak DO ( Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.)
Abstract	The modulation of cytoplasmic free Ca(2+) concentration ([Ca(2+)]i) is a universal intracellular signaling pathway that regulates numerous cellular physiological processes. Ubiquitous intracellular Ca(2+)-release channels localized to the endoplasmic/sarcoplasmic reticulum-inositol 1,4,5-trisphosphate receptor (InsP3R) and ryanodine receptor (RyR) channels-play a central role in [Ca(2+)]i signaling in all animal cells. Despite their intracellular localization, electrophysiological studies of the single-channel permeation and gating properties of these Ca(2+)-release channels using the powerful patch-clamp approach have been possible by application of this technique to isolated nuclei because the channels are present in membranes of the nuclear envelope. Here we provide a concise description of how nuclear patch-clamp experiments have been used to study single-channel properties of different InsP3R channels in the outer nuclear membrane. We compare this with other methods for studying intracellular Ca(2+) release. We also briefly describe application of the technique to InsP3R channels in the inner nuclear membrane and to channels in the outer nuclear membrane of HEK293 cells expressing recombinant RyR.
File Format	HTM / HTML
ISSN	19403402
e-ISSN	15596095
DOI	10.1101/pdb.top066217
Journal	Cold Spring Harbor Protocols
Issue Number	9
Volume Number	2013
Language	English
Publisher	Cold Spring Harbor Laboratory Press
Publisher Date	2013-09-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Clinical Laboratory Techniques Calcium Channels Physiology Calcium Metabolism Cytological Techniques Inositol 1,4,5-trisphosphate Receptors Nuclear Envelope Patch-clamp Techniques Cations, Divalent Cell Line
Content Type	Text
Resource Type	Article
Subject	Biochemistry, Genetics and Molecular Biology

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