Characterization of signaling pathways of P(2Y) and P(2U) purinoceptors in bovine pulmonary artery endothelial cells

B. C. Chen, C. M. Lee, Y. T. Lee, Wan Wan Lin

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The actions of ATP on the endothelium are mediated by P2 purinoceptors. We have shown that P(2Y) and P(2U) purinoceptors coexist in bovine pulmonary artery endothelial cells (CPAE), where they induce phosphoinositide (Pl) turnover and Ca2+ mobilization. The relative order of potency (based on the threshold concentration) of nucleotide analogues (1-100 μM) in stimulating the accumulation of inositol phosphate (IP) was 2-methylthio-ATP (2MeSATP) = 2-methylthio-ADP (2MeSADP) ≤ 2C1ATP > UTP = ATP = ADP. α,β-methylene ATP, β,γ-methylene ATP, UDP, adenosine-5'-tetraphospho-5'-adenosine, and adenosine-5'-pentaphospho-5'-adenosine had no effect at concentrations as high as 100 μM. At maximal concentrations, the IP responses to 2MeSATP and UTP were additive, whereas those to ATP and either 2MeSATP or UTP were not. Moreover, the maximal response to 2MeSADP was additive to that to UTP but not to that of 2MeSATP. Pretreatment with pertussis toxin slightly inhibited 2MeSATP-and UTP-stimulated IP generation by 15%. Under Ca2+-free conditions, UTP-induced IP formation was inhibited more markedly than that induced by 2MeSATP. Short-term treatment of the cells with phorbol 12- myristate-13-acetate (PMA) resulted in a dose-dependent inhibition of 2MeSATP-induced IP formation greater and more sensitive than that induced by UTP; similar results were obtained for the sensitivity of inhibition by suramin and reactive blue. Stimulation of the cells with either 2MeSATP or UTP induced a rapid increase in intracellular Ca2+ level, followed by a slow decrease to basal levels, followed by Ca2+ level oscillation. In the absence of extracellular Ca2+, [Ca2+](i) responses were quantitatively less and did not show the slow phase and oscillation. Together these results suggest that both P(2Y) and P(2U) purinoceptors are expressed in bovine pulmonary artery endothelial cells and are coupled to phospholipase C (PLC) activation and Ca2+ mobilization through pertussis toxin-insensitive G proteins.

Original languageEnglish
Pages (from-to)192-199
Number of pages8
JournalJournal of Cardiovascular Pharmacology
Volume28
Issue number2
DOIs
Publication statusPublished - 1996
Externally publishedYes

Fingerprint

Purinergic Receptors
Uridine Triphosphate
Pulmonary Artery
Endothelial Cells
Inositol Phosphates
Adenosine Triphosphate
Pertussis Toxin
Adenosine
Reactive Inhibition
2-methylthio-ATP
Purinergic P2 Receptors
Suramin
Uridine Diphosphate
Type C Phospholipases
Phosphatidylinositols
GTP-Binding Proteins
Adenosine Diphosphate
Endothelium
Acetates
Nucleotides

Keywords

  • Ca mobilization
  • P Receptor subtypes
  • Phosphoinositide turnover
  • Pulmonary artery endothelial cells

ASJC Scopus subject areas

  • Pharmacology
  • Cardiology and Cardiovascular Medicine

Cite this

Characterization of signaling pathways of P(2Y) and P(2U) purinoceptors in bovine pulmonary artery endothelial cells. / Chen, B. C.; Lee, C. M.; Lee, Y. T.; Lin, Wan Wan.

In: Journal of Cardiovascular Pharmacology, Vol. 28, No. 2, 1996, p. 192-199.

Research output: Contribution to journalArticle

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PY - 1996

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AB - The actions of ATP on the endothelium are mediated by P2 purinoceptors. We have shown that P(2Y) and P(2U) purinoceptors coexist in bovine pulmonary artery endothelial cells (CPAE), where they induce phosphoinositide (Pl) turnover and Ca2+ mobilization. The relative order of potency (based on the threshold concentration) of nucleotide analogues (1-100 μM) in stimulating the accumulation of inositol phosphate (IP) was 2-methylthio-ATP (2MeSATP) = 2-methylthio-ADP (2MeSADP) ≤ 2C1ATP > UTP = ATP = ADP. α,β-methylene ATP, β,γ-methylene ATP, UDP, adenosine-5'-tetraphospho-5'-adenosine, and adenosine-5'-pentaphospho-5'-adenosine had no effect at concentrations as high as 100 μM. At maximal concentrations, the IP responses to 2MeSATP and UTP were additive, whereas those to ATP and either 2MeSATP or UTP were not. Moreover, the maximal response to 2MeSADP was additive to that to UTP but not to that of 2MeSATP. Pretreatment with pertussis toxin slightly inhibited 2MeSATP-and UTP-stimulated IP generation by 15%. Under Ca2+-free conditions, UTP-induced IP formation was inhibited more markedly than that induced by 2MeSATP. Short-term treatment of the cells with phorbol 12- myristate-13-acetate (PMA) resulted in a dose-dependent inhibition of 2MeSATP-induced IP formation greater and more sensitive than that induced by UTP; similar results were obtained for the sensitivity of inhibition by suramin and reactive blue. Stimulation of the cells with either 2MeSATP or UTP induced a rapid increase in intracellular Ca2+ level, followed by a slow decrease to basal levels, followed by Ca2+ level oscillation. In the absence of extracellular Ca2+, [Ca2+](i) responses were quantitatively less and did not show the slow phase and oscillation. Together these results suggest that both P(2Y) and P(2U) purinoceptors are expressed in bovine pulmonary artery endothelial cells and are coupled to phospholipase C (PLC) activation and Ca2+ mobilization through pertussis toxin-insensitive G proteins.

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KW - P Receptor subtypes

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