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J. Biol. Chem., Vol. 275, Issue 22, 16626-16631, June 2, 2000
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From the Department of Physiology and Pharmacology, University of
Strathclyde, Glasgow G4 ONR and Pituitary adenylyl cyclase-activating
peptide (PACAP) stimulates calcium transients and catecholamine
secretion in adrenal chromaffin and PC12 cells. The PACAP type 1 receptor in these cells couples to both adenylyl cyclase and
phospolipase C pathways, but although phospolipase C has been
implicated in the response to PACAP, the role of adenylyl cyclase is
unclear. In this study, we show that PACAP38 stimulates
Ca2+ influx in PC12 cells by activating a cation
current that depends upon the dual activation of both the PLC and
adenylyl cyclase signaling pathways but does not involve protein kinase
C. In activating the current, PACAP38 has to overcome an inhibitory
effect of Ras. Thus, in cells expressing a dominant negative form of
Ras (PC12asn17-W7), PACAP38 induced larger, more rapidly activating
currents. This effect of Ras could be overidden by intracellular
guanosine-5'-O-3-(thio)triphosphate (GTP
Pituitary Adenylyl Cyclase-activating Peptide Activates Multiple
Intracellular Signaling Pathways to Regulate Ion Channels in PC12
Cells*
,§, and Division of Biochemistry
and Molecular Biology and Department of Medicine and Therapeutics,
University of Glasgow, Glasgow G12 8QQ, United Kingdom
S), suggesting
that it was mediated by inhibition of downstream G proteins. Ras may
also inhibit the current through a G protein-independent mechanism,
because cAMP analogues activated the current in PC12asn17-W7 cells,
provided GTPS was present, but not in PC12 cells expressing wild
type Ras. We conclude that coupling of PACAP to both adenylyl cyclase
and phospholipase C is required to activate Ca2+ influx in
PC12 cells and that tonic inhibition by Ras delays and limits the response.
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
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