Chronic hypoxia up-regulates alpha1H T-type channels and low-threshold catecholamine secretion in rat chromaffin cells

J Physiol. 2007 Oct 1;584(Pt 1):149-65. doi: 10.1113/jphysiol.2007.132274. Epub 2007 Aug 9.

Abstract

alpha(1H) T-type channels recruited by beta(1)-adrenergic stimulation in rat chromaffin cells (RCCs) are coupled to fast exocytosis with the same Ca(2+) dependence of high-threshold Ca(2+) channels. Here we show that RCCs exposed to chronic hypoxia (CH) for 12-18 h in 3% O(2) express comparable densities of functional T-type channels that depolarize the resting cells and contribute to low-voltage exocytosis. Following chronic hypoxia, most RCCs exhibited T-type Ca(2+) channels already available at -50 mV with the same gating, pharmacological and molecular features as the alpha(1H) isoform. Chronic hypoxia had no effects on cell size and high-threshold Ca(2+) current density and was mimicked by overnight incubation with the iron-chelating agent desferrioxamine (DFX), suggesting the involvement of hypoxia-inducible factors (HIFs). T-type channel recruitment occurred independently of PKA activation and the presence of extracellular Ca(2+). Hypoxia-recruited T-type channels were partially open at rest (T-type 'window-current') and contributed to raising the resting potential to more positive values. Their block by 50 microm Ni(2+) caused a 5-8 mV hyperpolarization. The secretory response associated with T-type channels could be detected following mild cell depolarizations, either by capacitance increases induced by step depolarizations or by amperometric current spikes induced by increased [KCl]. In the latter case, exocytotic bursts could be evoked even with 2-4 mm KCl and spike frequency was drastically reduced by 50 microm Ni(2+). Chronic hypoxia did not alter the shape of spikes, suggesting that hypoxia-recruited T-type channels increase the number of secreted vesicles at low voltages, without altering the mechanism of catecholamine release and the quantal content of released molecules.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism
  • Calcium Channels, T-Type / metabolism*
  • Catecholamines / metabolism*
  • Cells, Cultured
  • Chromaffin Cells / metabolism*
  • Deferoxamine
  • Exocytosis / physiology*
  • Hypoxia / metabolism*
  • Hypoxia-Inducible Factor 1 / metabolism
  • Membrane Potentials / physiology
  • Potassium Chloride / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Siderophores
  • Time Factors
  • Up-Regulation

Substances

  • Calcium Channels, T-Type
  • Catecholamines
  • Hypoxia-Inducible Factor 1
  • Siderophores
  • Potassium Chloride
  • Deferoxamine
  • Calcium