Nucleotide-regulated calcium signaling in lung fibroblasts and epithelial cells from normal and P2Y2 receptor (-/-) mice

L. Homolya, William C. Watt, Eduardo R. Lazarowski, Beverly H. Koller, Richard C. Boucher

Research output: Contribution to journalArticle

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Abstract

To test for the role of the P2Y2 receptor(P2Y2-R) in the regulation of nucleotide-promoted Ca2+ signaling in the lung, we generated P2Y2-R- deficient (P2Y2-R(-/-)) mice and measured intracellular Ca2+(i) responses (ΔCa2+(i)) to nucleotides in cultured lung fibroblasts and nasal and tracheal epithelial cells from wild type and P2Y2-R(-/-) mice. In the wild type fibroblasts, the rank order of potencies for nucleotide-induced ΔCa2+(i) was as follows: UTP ≥ ATP >> ADP > UDP. The responses induced by these agonists were completely absent in the P2Y2R(-/-) fibroblasts. Inositol phosphate responses paralleled those of ACa2+(i) in both groups. ATP and UTP also induced Ca2+(i) responses in wild type airway epithelial cells. In the P2Y2-R(-/-) airway epithelial cells, UTP was ineffective. A small fraction (25%) of the ATP response persisted. Adenosine and α,β- methylene ATP were ineffective, and ATP responses were not affected by adenosine deaminase or by removal of extracellular Ca2+, indicating that neither P1 nor P2X receptors mediated this residual ATP response. In contrast, 2-methylthio-ADP promoted a substantial Ca2+(i) response in P2Y2-R(-/-) cells, which was inhibited by the P2Y1 receptor antagonist adenosine 3'-5'-diphosphate. These studies demonstrate that P2Y2-R is the dominant purinoceptor in airway epithelial cells, which also express a P2Y1 receptor, and that the P2Y2-R is the sole purinergic receptor subtype mediating nucleotide-induced inositol lipid hydrolysis and Ca2+ mobilization in mouse lung fibroblasts.

Original languageEnglish
Pages (from-to)26454-26460
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number37
DOIs
Publication statusPublished - Sep 10 1999

Fingerprint

Purinergic P2Y2 Receptors
Calcium Signaling
Fibroblasts
Nucleotides
Epithelial Cells
Calcium
Adenosine Triphosphate
Lung
Uridine Triphosphate
Purinergic P2Y1 Receptors
Purinergic Receptors
Adenosine
Adenosine Deaminase
Inositol Phosphates
Uridine Diphosphate
Diphosphates
Inositol
Nose
Adenosine Diphosphate
Hydrolysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Nucleotide-regulated calcium signaling in lung fibroblasts and epithelial cells from normal and P2Y2 receptor (-/-) mice. / Homolya, L.; Watt, William C.; Lazarowski, Eduardo R.; Koller, Beverly H.; Boucher, Richard C.

In: Journal of Biological Chemistry, Vol. 274, No. 37, 10.09.1999, p. 26454-26460.

Research output: Contribution to journalArticle

Homolya, L. ; Watt, William C. ; Lazarowski, Eduardo R. ; Koller, Beverly H. ; Boucher, Richard C. / Nucleotide-regulated calcium signaling in lung fibroblasts and epithelial cells from normal and P2Y2 receptor (-/-) mice. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 37. pp. 26454-26460.
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