Adenosine receptor-mediated calcium mobilization in cortical collecting tubule cells

L. J. Arend, M. A. Burnatowska-Hledin, W. S. Spielman

Research output: Contribution to journalArticlepeer-review

Abstract

To investigate the cellular mechanisms underlying the epithelial actions of adenosine, we studied adenosine receptor-effector coupling in cultured rabbit cortical collecting tubule (RCCT) cells. We previously reported, in RCCT cells isolated by immunodissection, that a potent A2 adenosine analogue [5'-N-ethylcarboxamideadenosine (NECA)] stimulates cAMP production [effective concentration (EC 50)=1μM], and potent A1 analogues [N6-cyclohexyladenoisine) 50% (CHA) and R-N6-phenylisopropyladenosine (PIA)] inhibit basal and AVP-stimulated cAMP production (EC50 = 5 nM). The present study was undertaken to determine whether adenosine receptors in RCCT cells are also coupled to a signal transduction system leading to the mobilization of intracellular free calcium. RCCT cells were loaded with the fluorescent calcium indicator, fura-2, and were treated with the adenosine analogues NECA, CHA, and PIA. All three adenosine analogues produced dose-dependent (1 nM-0.1 mM), transient increases in intracellular calcium concentration with equal potency (EC50 = 0.5 μM). Chelation of extracellular calcium with ethyleneglycol-bis(β-aminoethyl ether)N,N,N',N' tetraacetic acid (EGTA) did not abolish the increase in calcium. The adenosine receptor antagonists, 1,3-diethyl-8-propylxanthine and 8-cyclopentyl-1,3-dipropylxanthine, and pretreatment of RCCT cells with pertussis toxin blocked the increase in calcium. These results demonstrate that RCCT cells have, in addition to adenosine receptors associated with the stimulation and inhibition of cAMP, a pertussis-toxin sensitive receptor system that leads to the mobilization of intracellular calcium.

Original languageEnglish (US)
Pages (from-to)24/5
JournalAmerican Journal of Physiology - Cell Physiology
Volume255
Issue number5
StatePublished - 1988

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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