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J Biol Chem, Vol. 274, Issue 47, 33348-33354, November 19, 1999
From the Department of Biophysical Genetics, Kanazawa University
Graduate School of Medicine, Kanazawa 920-8640, Japan
We examined the role of cyclic ADP-ribose
(cADP-ribose) as a second messenger downstream of adrenergic receptors
in the heart after excitation of sympathetic neurons. To address this
question, ADP-ribosyl cyclase activity was measured as the rate of
[3H]cADP-ribose formation from
[3H]NAD+ in a crude membrane fraction of rat
ventricular myocytes. Isoproterenol at 1 µM increased
ADP-ribosyl cyclase activity by 1.7-fold in ventricular muscle; this
increase was inhibited by propranolol. The stimulatory effect on the
cyclase was mimicked by 10 nM GTP and 10 µM
guanosine 5'-3-O-(thio)triphosphate, whereas 10 µM GTP inhibited the cyclase. Cholera toxin blocked the
activation of the cyclase by isoproterenol and GTP. The above effects
of isoproterenol and GTP in ventricular membranes were confirmed by
cyclic GDP-ribose formation fluorometrically. These results demonstrate
the existence of a signal pathway from 
-adrenergic receptors to
membrane-bound ADP-ribosyl cyclase via G protein in the ventricular
muscle cells and suggest that increased cADP-ribose synthesis is
involved in up-regulation of cardiac function by sympathetic stimulation.
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