The inositol 1,4,5-trisphosphate receptor (InsP3R) is an intracellular Ca2+-release channel localized in endoplasmic reticulum (ER) with a central role in complex Ca2+ signaling in most cell types. A family of InsP3Rs encoded by several genes has been identified with different primary sequences, subcellular locations, variable ratios of expression, and heteromultimer formation. This diversity suggests that cells require distinct InsP3Rs, but the functional correlates of this diversity are largely unknown. Lacking are single-channel recordings of the recombinant type 3 receptor (InsP3R-3), a widely expressed isoform also implicated in plasma membrane Ca2+ influx and apoptosis. Here, we describe functional expression and single-channel recording of recombinant rat InsP3R-3 in its native membrane environment. The approach we describe suggests a novel strategy for expression and recording of recombinant ER-localized ion channels in the ER membrane. Ion permeation and channel gating properties of the rat InsP3R-3 are strikingly similar to those of Xenopus type 1 InsP3R in the same membrane. Using two different two-electrode voltage clamp protocols to examine calcium store-operated calcium influx, no difference in the magnitude of calcium influx was observed in oocytes injected with rat InsP3R-3 cRNA compared with control oocytes. Our results suggest that if cellular expression of multiple InsP3R isoforms is a mechanism to modify the temporal and spatial features of [Ca2+]i signals, then it must be achieved by isoform-specific regulation or localization of various types of InsP3Rs that have relatively similar Ca2+ permeation properties.
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1 March 2000
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February 14 2000
Single-Channel Properties in Endoplasmic Reticulum Membrane of Recombinant Type 3 Inositol Trisphosphate Receptor
Don-On Daniel Mak,
Don-On Daniel Mak
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Sean McBride,
Sean McBride
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Viswanathan Raghuram,
Viswanathan Raghuram
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Yun Yue,
Yun Yue
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Suresh K. Joseph,
Suresh K. Joseph
cDepartment of Pathology, Thomas Jefferson University School of Medicine, Philadelphia, Pennsylvania 19107
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J. Kevin Foskett
J. Kevin Foskett
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
bFrom the Institute for Human Gene Therapy, University of Pennsylvania, Philadelphia, Pennsylvania 19104
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Don-On Daniel Mak
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Sean McBride
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Viswanathan Raghuram
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Yun Yue
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Suresh K. Joseph
cDepartment of Pathology, Thomas Jefferson University School of Medicine, Philadelphia, Pennsylvania 19107
J. Kevin Foskett
aFrom the Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
bFrom the Institute for Human Gene Therapy, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Abbreviations used in this paper: Ab-1, InsP3R-1–specific antibody; ER, endoplasmic reticulum; InsP3, inositol 1,4,5-trisphosphate; InsP3R, InsP3 receptor; SOC, store-operated Ca2+; TEV, two-electrode voltage-clamp.
Received:
November 10 1999
Revision Requested:
January 07 2000
Accepted:
January 18 2000
Online ISSN: 1540-7748
Print ISSN: 0022-1295
© 2000 The Rockefeller University Press
2000
The Rockefeller University Press
J Gen Physiol (2000) 115 (3): 241–256.
Article history
Received:
November 10 1999
Revision Requested:
January 07 2000
Accepted:
January 18 2000
Citation
Don-On Daniel Mak, Sean McBride, Viswanathan Raghuram, Yun Yue, Suresh K. Joseph, J. Kevin Foskett; Single-Channel Properties in Endoplasmic Reticulum Membrane of Recombinant Type 3 Inositol Trisphosphate Receptor. J Gen Physiol 1 March 2000; 115 (3): 241–256. doi: https://doi.org/10.1085/jgp.115.3.241
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