Functional mitochondria are required for O2 but not CO2 sensing in immortalized adrenomedullary chromaffin cells.
Catecholamine (CAT) release from adrenomedullary chromaffin cells (AMC) in response to stressors such as low O(2) (hypoxia) and elevated CO(2)/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 O(2) sensing. Wild-type MAH cells responded to both hypoxia and increased CO(2) (hypercapnia) with K(+) current inhibition and membrane depolarization. Additionally, these stimuli caused a rise in cytosolic Ca(2+) and CAT secretion, determined by fura-2 spectrofluorimetry and carbon fiber amperometry, respectively. In contrast, mitochondria-deficient (rho(0)) MAH cells were hypoxia insensitive, although responses to hypercapnia and expression of several markers, including carbonic anhydrase II, remained intact. Rotenone (1 microM), a mitochondrial complex I blocker known to mimic and occlude the effects of hypoxia in primary AMC, was effective in wild-type but not rho(0) MAH 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 CO(2) sensors; however, this property is independent of functional mitochondria.
Buttigieg
J
J
Department of Biology, McMaster University, Hamilton, Ontario, Canada.
Brown
S T
ST
Lowe
M
M
Zhang
M
M
Nurse
C A
CA
D000313:Adrenal Medulla / Q000166:cytology*; D000818:Animals; D002245:Carbon Dioxide / Q000378:metabolism*; D002395:Catecholamines / Q000378:metabolism; D015687:Cell Hypoxia; D002460:Cell Line; D019439:Chromaffin Cells / Q000187:drug effects / Q000378:metabolism*; D045222:Electron Transport Chain Complex Proteins / Q000037:antagonists & inhibitors; D008928:Mitochondria / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D051381:Rats; D012402:Rotenone / Q000494:pharmacology
D002395:Catecholamines; D045222:Electron Transport Chain Complex Proteins; D012402:Rotenone; D002245:Carbon Dioxide; D010100:Oxygen