The effects of cyclopiazonic acid on intracellular Ca2+ in aortic smooth muscle cells from DOCA-hypertensive rats

We tested the hypothesis that cyclopiazonic acid (CPA), an inhibitor of the sarcoplasmic reticulum (SR) Ca2+-ATPase, increases intracellular Ca2+ concentration ([Ca2+](i)) in aortic myocytes and that the increase in [Ca2+](i) is higher in aortic cells from deoxycorticosterone acetate (DOCA)-hypertensive rats. Male Sprague-Dawley rats, 250-300 g, underwent uninephrectomy, received a silastic implant containing DOCA (200 mg/kg) and had free access to water supplemented with 1.0% NaCl and 0.2% KCl. Control rats were also uninephrectomized, received normal tap water, but no implant. Intracellular Ca2+ measurements were performed in aortic myocytes isolated from normotensive (Systolic blood pressure = 120 +/- 3 mmHg; body weight = 478 +/- 7 g, N = 7) and DOCA-hypertensive rats (195 +/- 10 mmHg; 358 +/- 16 g, N = 7). The effects of CPA on resting [Ca2+](i) and on caffeine-induced increase in [Ca2+](i) after [Ca2+](i) depletion and reloading were compared in aortic cells from DOCA and normotensive rats. The phasic increase in [Ca2+](i) induced by 20 mM caffeine in Ca2+-free buffer was significantly higher in DOCA aortic cells (329 +/- 36 nM, N = 5) compared to that in normotensive cells (249 +/- 16 nM, N = 7, P<0.05). CPA (3 mu M) inhibited caffeine-induced increases in [Ca2+](i) in both groups. When the cells were placed in normal buffer (1.6 mM Ca2+, loading period), after treatment with Ca2+-free buffer (depletion period), an increase in [Ca2+](i) was observed in DOCA aortic cells (45 +/- 11 nM, N = 5) while no changes were observed in normotensive cells. CPA (3 mu M) potentiated the increase in [Ca2+](i) (122 +/- 30 nM, N = 5) observed in DOCA cells during the loading period while only a modest increase in [Ca2+](i) (23 +/- 10 nM, N = 5) was observed in normotensive cells. CPA-induced increase in [Ca2+](i) did not occur in the absence of extracellular Ca2+ or in the presence of nifedipine. These data show that CPA induces Ca2+ influx in aorta from both normotensive and DOCA-hypertensive rats. However, the increase in [Ca2+](i) is higher in DOCA aortic cells possibly due to an impairment in the mechanisms that control [Ca2+](i). The large increase in [Ca2+](i) in response to caffeine in DOCA cells probably reflects a greater storage of Ca2+ in the SR.