Adenosine-sensitive phosphoinositide turnover in a newly established renal cell line

Lois J Arend, J. S. Handler, J. S. Rhim, F. Gusovsky, W. S. Spielman

Research output: Contribution to journalArticle

Abstract

To aid in characterizing adenosine receptors in renal cells, primary cultures of rabbit cortical collecting tubule (RCCT) cells were infected with an adenovirus 12-simian virus 40 hybrid, resulting in a continuous cell line. The cells, designated RCCT-28A, retained their epithelial morphology and reacted with a monoclonal antibody specific for rabbit collecting tubule. Adenosine 3',5'-cyclic monophosphate (cAMP) accumulation was stimulated by vasopresin (AVP), isoproterenol, prostaglandin E2 (PGE2), calcitonin, parathyroid hormone, and a potent adenosine A1- and A2-receptor agonist, 5'-N-ethylcarboxamidoadenosine (NECA). A more selective adenosine A1-receptor agonist, N6-cyclohexyladenosine (CHA) inhibited basal and AVP-stimulated cAMP accumulation. Cytosolic free calcium was transiently elevated by bradykinin, PGE2, NECA, and CHA. To examine the mechanism by which adenosine analogues increase intracellular free calcium, phosphoinositide (PI) turnover was assessed in the 28A cells after labeling with myo-[3H]inositol. NECA and CHA increased [3H]inositol phosphate formation with an approximate half-maximal effective concentration of 0.1 μM for both analogues. The increase in PI turnover was blocked by the selective adenosine A1-receptor antagonist, 8-cyclopentyl-1,3-dipropylxanthine and pretreatment of the 28A cells with pertussis toxin. These results suggest that adenosine analogues increase cytosolic free calcium by stimulating PI turnover.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Fluid and Electrolyte Physiology
Volume256
Issue number6
StatePublished - 1989

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Adenosine-5'-(N-ethylcarboxamide)
Phosphatidylinositols
Adenosine
Adenosine A1 Receptor Agonists
Kidney
Cell Line
Rabbits
Calcium
Dinoprostone
Adenosine A2 Receptor Agonists
Adenosine A1 Receptor Antagonists
Purinergic P1 Receptors
Inositol Phosphates
Primary Cell Culture
Simian virus 40
Pertussis Toxin
Calcitonin
Bradykinin
Inositol
Parathyroid Hormone

ASJC Scopus subject areas

  • Physiology

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Adenosine-sensitive phosphoinositide turnover in a newly established renal cell line. / Arend, Lois J; Handler, J. S.; Rhim, J. S.; Gusovsky, F.; Spielman, W. S.

In: American Journal of Physiology - Renal Fluid and Electrolyte Physiology, Vol. 256, No. 6, 1989.

Research output: Contribution to journalArticle

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