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J Biol Chem, Vol. 273, Issue 17, 10402-10410, April 24, 1998
Characterization of a Ca2+ Release-activated
Nonselective Cation Current Regulating Membrane Potential and
[Ca2+]i Oscillations in Transgenically
Derived -Cells
Michael W.
Roe ,
Jennings F.
Worley III§,
Feng
Qian¶,
Natalia
Tamarina ,
Anshu A.
Mittal ,
Flora
Dralyuk ,
Nathaniel T.
Blair ,
Robert J.
Mertz§,
Louis H.
Philipson , and
Iain D.
Dukes§
From the Departments of Medicine and
¶ Pharmacology and Physiology, University of Chicago, Chicago,
Illinois 60637 and the § Department of Cell Physiology,
Glaxo Wellcome Research Institute,
Research Triangle Park, North Carolina 27709
Although stimulation of insulin secretion by
glucose is regulated by coupled oscillations of membrane potential and
intracellular Ca2+ ([Ca2+]i),
the membrane events regulating these oscillations are incompletely
understood. In the presence of glucose and tetraethylammonium, transgenically derived -cells ( TC3-neo) exhibit coupled voltage and [Ca2+]i oscillations strikingly similar to
those observed in normal islets in response to glucose. Using these
cells as a model system, we investigated the membrane conductance
underlying these oscillations. Alterations in delayed
rectifier or Ca2+-activated K+ channels
were excluded as a source of the conductance oscillations, as they
are completely blocked by tetraethylammonium. ATP-sensitive K+ channels were also excluded, since the ATP-sensitive
K+ channel blocker tolbutamide substituted for glucose in
inducing [Ca2+]i oscillations. Thapsigargin,
which depletes intracellular Ca2+ stores, and maitotoxin,
an activator of nonselective cation channels, both converted the
glucose-dependent [Ca2+]i
oscillations into a sustained elevation. On the other hand, both SKF
96365, a blocker of Ca2+ store-operated channels, and
external Na+ removal suppressed the glucose-stimulated
[Ca2+]i oscillations. Maitotoxin activated a
nonselective cation current in TC3 cells that was attenuated by
removal of extracellular Na+ and by SKF 96365, in the same
manner to a current activated in mouse -cells following depletion of
intracellular Ca2+ stores. Currents similar to these are
produced by the mammalian trp-related channels, a gene
family that includes Ca2+ store-operated channels and
inositol 1,4,5-trisphosphate-activated channels. We found several of
the trp family genes were expressed in TC3 cells by
reverse transcriptase polymerase chain reaction using specific primers,
but by Northern blot analysis, mtrp-4 was the predominant
message expressed. We conclude that a conductance underlying
glucose-stimulated oscillations in -cells is provided by a
Ca2+ store depletion-activated nonselective cation current,
which is plausibly encoded by homologs of trp genes.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 1998 by the American Society for Biochemistry and Molecular Biology.
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