© 1997 American Heart Association, Inc.
Articles |
Actions of cADP-Ribose and Its Antagonists on Contraction in Guinea Pig Isolated Ventricular Myocytes
Influence of Temperature
From the Department of Pharmacology, University of Oxford (UK).
Correspondence to Dr D.A. Terrar, Department of Pharmacology, University of Oxford, Mansfield Road, Oxford OX1 3QT, UK. E-mail derek.terrar{at}pharm.ox.ac.uk
Abstract Although it is becoming widely accepted that
cADP-ribose (cADPR) can regulate calcium release from the endoplasmic
reticulum in sea urchin eggs and in a variety of mammalian cell types,
it remains controversial whether this substance might influence calcium
release during excitation-contraction coupling in cardiac muscle. We
have investigated possible actions of cADPR in intact cells isolated
from guinea pig ventricle, paying particular attention to the possible
influence of temperature. At 36°C, myocyte contraction was influenced
by cytosolic application of cADPR in a concentration-dependent manner
(showing an 
30% increase in contraction with 5 µmol/L cADPR
applied via a patch pipette in myocytes stimulated to fire action
potentials at 1 Hz). Calcium transients measured with fura 2 were also
increased by 5 µmol/L cADPR. Antagonists of cADPR
reduced contraction at 36°C (by 35% with either 50 µmol/L
8-Br-cADPR or 5 µmol/L 8-amino-cADPR applied via the patch
pipette). At room temperature (20°C to 24°C), no significant
effects on contraction were detected with either cADPR or its
antagonists. At 36°C, treatment of the cells with a
mixture of 2 µmol/L ryanodine and 1 µmol/L thapsigargin
to suppress function of the sarcoplasmic reticulum stores of calcium
prevented the action of 5 µmol/L cADPR applied via a patch
pipette. These observations are consistent with an action of
cytosolic cADPR to enhance calcium-induced calcium release from the
sarcoplasmic reticulum in guinea pig ventricular myocytes
at 36°C. The observed influence of temperature under the conditions
of our experiments is one factor that might help to account for failure
to detect actions of cADPR and its analogues in some previous studies.
Key Words: cADP-ribose • ryanodine receptor • cardiac muscle • sarcoplasmic reticulum
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