TY - JOUR
T1 - Measurement of adenylylcyclase activity in the AV nodal region of the canine heart
T2 - Evidence for inhibition by adenosine and acetylcholine
AU - Sugiyama, Atsushi
AU - McKnite, Scott
AU - Adkisson, Wayne O
AU - Lurie, Keith G
PY - 1997/6
Y1 - 1997/6
N2 - Although it is essential to cardiac conduction, little is known about the biochemistry underlying postreceptor adrenergic, cholinergic and purinergic processes in the AV node. To study these mechanisms, we adapted a new and highly sensitive fluorometric assay for cyclic adenosine monophosphate (AMP) to characterize regional adenylylcyclase activity (cyclic AMP production in pmol/min/mg of protein) in membrane preparations made from 20-50 pieces of freeze-dried, 20-μn thick, microdissected samples of tissue from canine tight atrium, the AV nodal region, and left ventricle. Basal and NaF-stimulated adenylylcyclase activity (mean ± SEM, n = 6) were 7.2 ± 0.4 and 72.4 ± 7.5 in atrial, 15.6 ± 1.3 and 58.8 ± 4.7 in AV nodal, and 6,4 ± 0.9 and 66.7 ± 5.0 in ventricular tissues, respectively. Isoproterenol (10-7-10-4 M) increased adenylylcyclase activity in a dose-dependent fashion in three different regions. The isoproterenol (10-6 M)-stimulated adenylylcy]clase activity (n=6) was 14.4 ± 1.3 in atrial, 21.9 ± 1.6 in AV nodal and 13.4 ± 1.4 in ventricular tissues. Adenosine (10-3 M) and carbachol (10-5 M) inhibited isoproterenol (10-6 M)-stimulated adenylylcyclase activity to 10.1 ± 1.1, 12.9 ± 1.3 in atrial, 15.1 ± 1.6, 15.5 ± 1.2 in AV nodal, and 7.5 ± 0.7, 11.9 ± 1.2 in ventricular tissues, respectively. The results demonstrate that there are regional differences in adenylylcyclase activity under basal conditions and after adrenergic, purinergic, and cholinergic stimulation in the heart. Unlike adenosine, the inhibitory effects of cholinergic stimulation appear to be more specific for the AV node.
AB - Although it is essential to cardiac conduction, little is known about the biochemistry underlying postreceptor adrenergic, cholinergic and purinergic processes in the AV node. To study these mechanisms, we adapted a new and highly sensitive fluorometric assay for cyclic adenosine monophosphate (AMP) to characterize regional adenylylcyclase activity (cyclic AMP production in pmol/min/mg of protein) in membrane preparations made from 20-50 pieces of freeze-dried, 20-μn thick, microdissected samples of tissue from canine tight atrium, the AV nodal region, and left ventricle. Basal and NaF-stimulated adenylylcyclase activity (mean ± SEM, n = 6) were 7.2 ± 0.4 and 72.4 ± 7.5 in atrial, 15.6 ± 1.3 and 58.8 ± 4.7 in AV nodal, and 6,4 ± 0.9 and 66.7 ± 5.0 in ventricular tissues, respectively. Isoproterenol (10-7-10-4 M) increased adenylylcyclase activity in a dose-dependent fashion in three different regions. The isoproterenol (10-6 M)-stimulated adenylylcy]clase activity (n=6) was 14.4 ± 1.3 in atrial, 21.9 ± 1.6 in AV nodal and 13.4 ± 1.4 in ventricular tissues. Adenosine (10-3 M) and carbachol (10-5 M) inhibited isoproterenol (10-6 M)-stimulated adenylylcyclase activity to 10.1 ± 1.1, 12.9 ± 1.3 in atrial, 15.1 ± 1.6, 15.5 ± 1.2 in AV nodal, and 7.5 ± 0.7, 11.9 ± 1.2 in ventricular tissues, respectively. The results demonstrate that there are regional differences in adenylylcyclase activity under basal conditions and after adrenergic, purinergic, and cholinergic stimulation in the heart. Unlike adenosine, the inhibitory effects of cholinergic stimulation appear to be more specific for the AV node.
KW - Acetylcholine
KW - Adenosine
KW - Adenosine 3':5'
KW - Adenylylcyclase
KW - Atrioventricular node
KW - Isoproterenol
KW - Monophosphate
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U2 - 10.1097/00005344-199706000-00005
DO - 10.1097/00005344-199706000-00005
M3 - Article
C2 - 9234653
AN - SCOPUS:0030852285
SN - 0160-2446
VL - 29
SP - 734
EP - 739
JO - Journal of Cardiovascular Pharmacology
JF - Journal of Cardiovascular Pharmacology
IS - 6
ER -