Effect of extracellular calcium depletion on the two-step ANP secretion in perfused rabbit atria

Employing isolated perfused rabbit atrial model we have found that stretch-activated atrial natriuretic peptide (ANP) secretion takes place in two steps: release of ANP from myocytes into surrounding intercellular space, and then translocation of the released ANP into atrial lumen along with the extracellular fluid (ECF) translocated upon releasing the stretch. Ca2+, one of the most important factors regulating secretory processes, has been reported by many workers to have influence on the ANP secretion, but at large variance. In the present study, therefore, we undertook to clarify the influence of Ca2+ depletion on ANP secretion and further to define which of the two steps is affected by Ca2+ removal. Extracellular Ca2+ depletion resulted in increased basal secretion and in accentuated secretion of immunoreactive (ir) ANP in response to mechanical stimuli. The translocation of ECF increased in response to atrial stretch-and-release, but it was not affected by Ca2+ depletion. The irANP concentration in the extracellular space calculated as the amount of irANP secreted in relation to the ECF translocated significantly increased when extracellular Ca2+ was depleted. These results indicate that extracellular Ca2+ depletion accentuates the stretch-induced ANP secretion through the augmentation of ANP release into the interstitium without changes in the ECF translocation.