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J. Biol. Chem., Vol. 266, Issue 24, 15910-15916, 08, 1991
CA Sei, CE Irons, AB Sprenkle, PM McDonough, JH Brown and CC Glembotski
It has been shown recently that alpha-adrenergic agonists can stimulate
atrial natriuretic factor (ANF) expression in ventricular cardiac myocytes;
however, little is known about the intracellular signals mediating this
activation. The present study focused on the potential roles of
calcium-regulated kinases and calcium influx in the alpha- adrenergic
stimulation of ANF gene expression in ventricular myocardial cell cultures.
Myocardial cells maintained for 48 h in serum-free medium supplemented with
phenylephrine (PE) possessed up to 15-fold higher levels of ANF peptide and
ANF mRNA than control cells. The removal of PE, or the addition of
nifedipine, resulted in a rapid decline in ANF expression, suggesting that
the sustained elevation of some intracellular messenger (e.g. calcium
and/or phospholipid hydrolysis products) was required for the adrenergic
response. The calcium channel agonist BAY K 8644 was capable of increasing
ANF expression in a nifedipine-sensitive manner; however, unlike PE, it did
not stimulate phosphoinositide hydrolysis. The protein kinase C inhibitor,
H7, caused an approximate 75% reduction in PE-stimulated ANF expression,
but had no effect on BAY K-stimulated expression. W7, a calcium/calmodulin
inhibitor, completely blocked the effects of both PE and BAY K 8644. The
addition of either H7 or W7 24 h after the PE addition resulted in a
decline of ANF expression. These results indicate that alpha-adrenergic
agonists augment ANF gene expression through at least two pathways, one
that is H7-sensitive, perhaps involving the sustained activation of protein
kinase C, and the other that is W7-sensitive, perhaps involving the
sustained activation of calmodulin-regulated kinases. Further, it appears
that BAY K 8644- mediated increases in ANF expression are independent of
protein kinase C activation and dependent on calmodulin-regulated events.
The alpha-adrenergic stimulation of atrial natriuretic factor expression in cardiac myocytes requires calcium influx, protein kinase C, and calmodulin-regulated pathways
Department of Biology, San Diego State University, California 92182.
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