title: Adrenocorticotropin evokes transient elevations in intracellular free calcium ([Ca2+]i) and increases basal [Ca2+]i in resting chondrocytes through a phospholipase C-dependent mechanism.
abstract: Both clinical and in vitro evidence points to the involvement of the melanocortin peptide, ACTH, in the terminal differentiation of chondrocytes. Terminal differentiation along the endochondral pathway is responsible for linear growth, but also plays a role in osteoarthritic cartilage degeneration. Chondrocyte terminal differentiation is associated with an incremental increase in chondrocyte basal intracellular free calcium ([Ca(2+)](i)), and ACTH agonism of melanocortin receptors is known to mobilize [Ca(2+)](i.) Using differentiated resting chondrocytes highly expressing type II collagen and aggrecan, we examined the influence of both ACTH and dexamethasone treatment on matrix gene transcription and [Ca(2+)](i). Resting chondrocytes treated concurrently with dexamethasone and ACTH expressed matrix gene transcripts in a pattern consistent with that of rapid terminal differentiation. Using the fluorescent Ca(2+) indicator, fura-2, we determined that ACTH evokes transient increases in [Ca(2+)](i) and elevates basal Ca(2+) levels in resting chondrocytes. The transient increases were initiated intracellularly, were abrogated by the phospholipase C-specific inhibitor, U73122, and were partly attenuated by myo-inositol 1,4,5-triphosphate receptor inhibition via 10 mm caffeine. The initial intracellular release also resulted in store-operated calcium entry, presumably through store-operated channels. Dexamethasone priming increased both the initial ACTH-evoked [Ca(2+)](i) release and the subsequent store-operated calcium entry. These data demonstrate roles for ACTH and glucocorticoid in the regulation of chondrocyte terminal differentiation. Because the actions of ACTH are mediated through known G protein-coupled receptors, the melanocortin receptors, these data may provide a new therapeutic target in the treatment of growth deficiencies and cartilage degeneration.
authors.affiliation: Department of Medicine, Winthrop University Hospital, Mineola, New York 11501, USA. jevans@winthrop.org
authors.forename: Jodi F
authors.initials: JF
authors.lastname: Evans
authors.forename: Chwan-L
authors.initials: CL
authors.lastname: Shen
authors.forename: Simcha
authors.initials: S
authors.lastname: Pollack
authors.forename: John F
authors.initials: JF
authors.lastname: Aloia
authors.forename: James K
authors.initials: JK
authors.lastname: Yeh
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000502:physiology; D002460:Cell Line; D019902:Chondrocytes / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004351:Drug Resistance; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005938:Glucocorticoids / Q000494:pharmacology; D053496:Inositol 1,4,5-Trisphosphate Receptors; D007425:Intracellular Membranes / Q000378:metabolism*; D009074:Melanocyte-Stimulating Hormones / Q000008:administration & dosage / Q000494:pharmacology; D009994:Osmolar Concentration; D051381:Rats; D017207:Rats, Sprague-Dawley; D018043:Receptors, Corticotropin / Q000037:antagonists & inhibitors; D018160:Receptors, Cytoplasmic and Nuclear / Q000502:physiology; D019284:Thapsigargin / Q000494:pharmacology; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D010738:Type C Phospholipases / Q000502:physiology*
chemical_list: D015220:Calcium Channels; D004791:Enzyme Inhibitors; D005938:Glucocorticoids; D053496:Inositol 1,4,5-Trisphosphate Receptors; D018043:Receptors, Corticotropin; D018160:Receptors, Cytoplasmic and Nuclear; C103272:SHU 9119; D019284:Thapsigargin; D003907:Dexamethasone; D000324:Adrenocorticotropic Hormone; D009074:Melanocyte-Stimulating Hormones; D010738:Type C Phospholipases; D002118:Calcium