title: Ca2+ and K+ channels of normal human adrenal zona fasciculata cells: properties and modulation by ACTH and AngII.
abstract: In whole cell patch clamp recordings, we found that normal human adrenal zona fasciculata (AZF) cells express voltage-gated, rapidly inactivating Ca(2+) and K(+) currents and a noninactivating, leak-type K(+) current. Characterization of these currents with respect to voltage-dependent gating and kinetic properties, pharmacology, and modulation by the peptide hormones adrenocorticotropic hormone (ACTH) and AngII, in conjunction with Northern blot analysis, identified these channels as Cav3.2 (encoded by CACNA1H), Kv1.4 (KCNA4), and TREK-1 (KCNK2). In particular, the low voltage-activated, rapidly inactivating and slowly deactivating Ca(2+) current (Cav3.2) was potently blocked by Ni(2+) with an IC50 of 3 µM. The voltage-gated, rapidly inactivating K(+) current (Kv1.4) was robustly expressed in nearly every cell, with a current density of 95.0 ± 7.2 pA/pF (n = 64). The noninactivating, outwardly rectifying K(+) current (TREK-1) grew to a stable maximum over a period of minutes when recording at a holding potential of -80 mV. This noninactivating K(+) current was markedly activated by cinnamyl 1-3,4-dihydroxy-α-cyanocinnamate (CDC) and arachidonic acid (AA) and inhibited almost completely by forskolin, properties which are specific to TREK-1 among the K2P family of K(+) channels. The activation of TREK-1 by AA and inhibition by forskolin were closely linked to membrane hyperpolarization and depolarization, respectively. ACTH and AngII selectively inhibited the noninactivating K(+) current in human AZF cells at concentrations that stimulated cortisol secretion. Accordingly, mibefradil and CDC at concentrations that, respectively, blocked Cav3.2 and activated TREK-1, each inhibited both ACTH- and AngII-stimulated cortisol secretion. These results characterize the major Ca(2+) and K(+) channels expressed by normal human AZF cells and identify TREK-1 as the primary leak-type channel involved in establishing the membrane potential. These findings also suggest a model for cortisol secretion in human AZF cells wherein ACTH and AngII receptor activation is coupled to membrane depolarization and the activation of Cav3.2 channels through inhibition of hTREK-1.
authors.affiliation: Department of Neuroscience, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA. enyeart.1@osu.edu
authors.forename: John J
authors.initials: JJ
authors.lastname: Enyeart
authors.forename: Judith A
authors.initials: JA
authors.lastname: Enyeart
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000293:Adolescent; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000328:Adult; D000804:Angiotensin II / Q000494:pharmacology*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020747:Calcium Channels, T-Type / Q000235:genetics / Q000378:metabolism*; D002648:Child; D005260:Female; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism; D015640:Ion Channel Gating; D051665:Kv1.4 Potassium Channel / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008297:Male; D008564:Membrane Potentials; D008875:Middle Aged; D026902:Potassium Channel Blockers / Q000494:pharmacology; D024683:Potassium Channels, Tandem Pore Domain / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D015383:Zona Fasciculata / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology
chemical_list: C479952:CACNA1H protein, human; D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; C496977:KCNA4 protein, human; D051665:Kv1.4 Potassium Channel; D026902:Potassium Channel Blockers; D024683:Potassium Channels, Tandem Pore Domain; C103742:potassium channel protein TREK-1; D000804:Angiotensin II; D000324:Adrenocorticotropic Hormone; D006854:Hydrocortisone