title: Effects of hypoxia and dithionite on catecholamine release from isolated type I cells of the rat carotid body.
abstract: 1. Amperometric recordings were conducted to investigate the ability of hypoxia and anoxia to evoke quantal catecholamine secretion from isolated type I cells of the rat carotid body. 2. Hypoxia (PO2 8-14 mmHg) consistently failed to evoke catecholamine secretion from type I cells, when cells were perfused either at room temperature (21-24 C) or at 35-37 C, and regardless of whether Hepes- or HCO3-/CO2-buffered solutions were used. 3. Elevating extracellular [K+] caused concentration-dependent secretion from individual type I cells, with a threshold concentration of approximately 25 mM. In the presence of this level of extracellular K+, hypoxia (PO2 8-14 mmHg) caused a marked enhancement of secretion which was fully blocked by 200 microM Cd2+, a non-specific blocker of voltage-gated Ca2+ channels. 4. Anoxia (N2-equilibrated solution containing 0.5 mM dithionite) evoked exocytosis from type I cells when extracellular [K+] was 5 mM. This secretion was completely inhibited by removal of extracellular Ca2+, but was not significantly affected by Cd2+ (200 microM), Ni2+ (2 mM), Zn2+ (1 mM) or nifedipine (2 microM). Secretion was also observed when 0.5 mM dithionite was added to air-equilibrated solutions. 5. Anoxia also evoked secretion from chemoreceptive phaeochromocytoma (PC12) cells, which was wholly Ca2+ dependent, but unaffected by Cd2+ (200 microM). 6. Our results suggest that hypoxia can evoke catecholamine secretion from isolated type I cells, but only in the presence of elevated extracellular [K+]. This may be due to the cells being relatively hyperpolarized following dissociation. In addition, we have shown that dithionite evokes catecholamine release regardless of PO2 levels, and this release is due mainly to an artefactual Ca2+ influx pathway activated in the presence of dithionite.
authors.affiliation: Institute for Cardiovascular Research, University of Leeds, Leeds LS2 9JT and Department of Pharmacology, University College London, London WC1E 6BT, UK.
authors.forename: E
authors.initials: E
authors.lastname: Carpenter
authors.forename: C J
authors.initials: CJ
authors.lastname: Hatton
authors.forename: C
authors.initials: C
authors.lastname: Peers
mesh_terms: D000818:Animals; D002104:Cadmium / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D020746:Calcium Channels, L-Type / Q000187:drug effects; D002344:Carotid Body / Q000166:cytology / Q000378:metabolism*; D002395:Catecholamines / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002469:Cell Separation; D004227:Dithionite / Q000494:pharmacology*; D004594:Electrophysiology / Q000379:methods; D005089:Exocytosis; D005110:Extracellular Space / Q000378:metabolism; D000860:Hypoxia / Q000378:metabolism*; D009994:Osmolar Concentration; D016716:PC12 Cells / Q000378:metabolism; D011188:Potassium / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature
chemical_list: D020746:Calcium Channels, L-Type; D002395:Catecholamines; D002104:Cadmium; D004227:Dithionite; D011188:Potassium; D002118:Calcium