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J Biol Chem, Vol. 275, Issue 19, 14691-14699, May 12, 2000
From the Departments of Capacitative Ca2+ entry
stimulates cAMP synthesis in mouse parotid acini, suggesting that one
of the Ca2+-sensitive adenylyl cyclases (AC1 or AC8) may
play an important role in the regulation of parotid function (Watson,
E. L., Wu, Z., Jacobson, K. L., Storm, D. R., Singh,
J. C., and Ott, S. M. (1998) Am. J. Physiol.
274, C557-C565). To evaluate the role of AC1 and AC8 in
Ca2+ stimulation of cAMP synthesis in parotid cells, acini
were isolated from AC1 mutant (AC1-KO) and AC8 mutant (AC8-KO) mice and
analyzed for Ca2+ stimulation of intracellular cAMP levels.
Although Ca2+ stimulation of intracellular cAMP levels in
acini from AC1-KO mice was indistinguishable from wild type mice, acini
from AC8-KO mice showed no Ca2+-stimulated cAMP
accumulation. This indicates that AC8, but not AC1, plays a major role
in coupling Ca2+ signals to cAMP synthesis in parotid
acini. Interestingly, treatment of acini from AC8-KO mice with agents,
i.e. carbachol and thapsigargin that increase intracellular
Ca2+, lowered cAMP levels. This decrease was dependent upon
Ca2+ influx and independent of phosphodiesterase
activation. Immunoblot analysis revealed that AC5/6 and AC3 are
expressed in parotid glands. Inhibition of calmodulin (CaM) kinase II
with KN-62, or inclusion of the CaM inhibitor, calmidazolium, did not
prevent agonist-induced inhibition of stimulated cAMP accumulation.
In vitro studies revealed that Ca2+,
independently of CaM, inhibited isoproterenol-stimulated AC. Data
suggest that agonist augmentation of stimulated cAMP levels is due to
activation of AC8 in mouse parotid acini, and strongly support a role
for AC5/6 in the inhibition of stimulated cAMP levels.
The Type 8 Adenylyl Cyclase Is Critical for Ca2+
Stimulation of cAMP Accumulation in Mouse Parotid Acini*
§¶,,,,, Oral Biology and
§ Pharmacology, University of Washington,
Seattle, Washington 98195
*
This work was supported by National Institutes of Health
Grants DE-05249 and NS-20498.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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