Voltage dependent calcium and potassium currents in Y-1 adrenocortical cells are unresponsive to ACTH.
In this report we use both whole cell and perforated patch clamp recording techniques to characterize calcium and potassium channels in Y-1 adrenocortical cells in order to assess their responsiveness to ACTH. Both transient and long-lasting components of an inward calcium current were identified which were similar to T and L-type Ca2+ currents. With Ba2+ as the charge carrier, the transient current activated at voltages more hyperpolarized than -50 mV with V1/2 for activation at -78.1 mV, and for steady state inactivation at -52.3 mV. The L-type current activated at -20 mV, with a V1/2 for activation at -29.9 mV and steady state inactivation at -44.2 mV. Under perforated patch conditions the response was shifted to more depolarized voltages. Both currents were responsive to agents which usually affect T- or L-type Ca2+ currents. The transient current was completely blocked by 50 microM lanthanum or 200 microM nickel and partially blocked by 300 mM amiloride. Cadmium (100 microM) and nifedipine (300 nM) completely blocked the long-lasting current while omega-conotoxin GVIA (1992 nM) inhibited the current by only 20-25%. The agonist, Bay K 8644 was stimulatory at 50 nM. Both transient and sustained outward potassium currents similar to A-type and delayed rectifier currents, respectively, were present. The transient current demonstrated fast activation at voltages more positive than -10 mV, inactivation with continued depolarization and steady state inactivation at V1/2 = -50 mV. The sustained current activated rapidly and had minimal inactivation with continued depolarization. The transient current was blocked by 5 mM 4AP and the sustained by 25 mM TEA. While Y-1 cells contain both calcium and potassium currents similar to those found in other adrenocortical cells, none of the currents were affected by ACTH or AII, secretagogues which stimulate steroidogenesis. These data, combined with the inability of both Ca2+ and K+ channel blockers to alter ACTH-induced steroidogenesis as reported earlier, suggests that neither calcium nor potassium currents are responsive to ACTH in Y-1 cells.
Coyne
M D
MD
Department of Biological Sciences, Wellesley College, MA 02181, USA.
Rodriguez
O
O
Wilson
Y
Y
Wang
G
G
Lemos
J R
JR
D000302:Adrenal Cortex / Q000166:cytology*; D000306:Adrenal Cortex Neoplasms; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000145:classification / Q000187:drug effects*; D004095:Dihydropyridines / Q000494:pharmacology; D004558:Electric Stimulation; D004594:Electrophysiology; D051379:Mice; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000494:pharmacology; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000145:classification / Q000187:drug effects*; D014407:Tumor Cells, Cultured; D020866:omega-Conotoxin GVIA
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D004095:Dihydropyridines; D010455:Peptides; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D000804:Angiotensin II; D000324:Adrenocorticotropic Hormone; D020866:omega-Conotoxin GVIA