Functional mitochondria are required for O2but not CO2sensing in immortalized adrenomedullary chromaffin cells

Abstract

Catecholamine (CAT) release from adrenomedullary chromaffin cells (AMC) in response to stressors such as low O2(hypoxia) and elevated CO2/H+is critical during adaptation of the newborn to extrauterine life. Using a surrogate model based on a v -myc immortalized adrenal chromaffin cell line (i.e., MAH cells), combined with genetic perturbation of mitochondrial function, we tested the hypothesis that functional mitochondria are required for O2sensing. Wild-type MAH cells responded to both hypoxia and increased CO2(hypercapnia) with K+current inhibition and membrane depolarization. Additionally, these stimuli caused a rise in cytosolic Ca2+and CAT secretion, determined by fura-2 spectrofluorimetry and carbon fiber amperometry, respectively. In contrast, mitochondria-deficient (ρ0) MAH cells were hypoxia insensitive, although responses to hypercapnia and expression of several markers, including carbonic anhydrase II, remained intact. Rotenone (1 μM), a mitochondrial complex I blocker known to mimic and occlude the effects of hypoxia in primary AMC, was effective in wild-type but not ρ0MAH cells. These data demonstrate that functional mitochondria are involved in hypoxia-sensing by adrenal chromaffin cells. We also show for the first time that, like their neonatal chromaffin cell counterparts, MAH cells are CO2sensors; however, this property is independent of functional mitochondria.