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Journal of Neuroscience, Vol 15, 2961-2971, Copyright © 1995 by Society for Neuroscience
ATP causes release of intracellular Ca2+ via the phospholipase C beta/IP3 pathway in astrocytes from the dorsal spinal cord
MW Salter and JL Hicks
Hospital for Sick Children, Department of Physiology, University of Toronto, Ontario, Canada.
Calcium signaling within astrocytes in the CNS may play a role comparable
to that of electrical signaling within neurons. ATP is a molecule known to
produce Ca2+ responses in astrocytes, and has been implicated as a mediator
of intercellular Ca2+ signaling in other types of nonexcitable cells. We
characterized the signal transduction pathway for ATP-evoked Ca2+ responses
in cultured astrocytes from the dorsal spinal cord. Nearly 100% of these
astrocytes respond to extracellularly applied ATP, which causes release of
Ca2+ from an intracellular pool that is sensitive to thapsigargin and
insensitive to caffeine. We found that intracellular administration of IP3
also caused release of Ca2+ from a thapsigargin-sensitive intracellular
pool, and that IP3 abolished the response to ATP. The ATP-evoked Ca2+
response was blocked by the IP3 receptor antagonist heparin, applied
intracellularly, but not by N-desulfated heparin, which is not an
antagonist at these receptors. The Ca2+ response caused by ATP was also
blocked by a phospholipase C inhibitor, U-73122, but not by its inactive
analog, U- 73343. Increases in [Ca2+]i were elicited by intracellular
application of activators of heterotrimeric G-proteins, GTP gamma S and
AIF4-. On the other hand, [Ca2+], was unaffected by a G-protein inhibitor,
GDP beta S, but it did abolish the Ca2+ response to ATP. Pretreating the
cultures with pertussis toxin did not affect responses to ATP. Our results
indicate that in astrocytes ATP-evoked release of intracellular Ca2+ is
mediated by IP3 produced as a result of activating phospholipase C coupled
to ATP receptors via a G-protein that is insensitive to pertussis toxin.
ATP is known to be released under physiological and pathological
circumstances, and therefore signaling via the PLC-IP3 pathway in
astrocytes is a potentially important mechanism by which ATP may play a
role in CNS function.
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