Stimulation of P2Y receptors activates c-fos gene expression and inhibits DNA synthesis in cultured cardiac fibroblasts

Jing Sheng Zheng, Lydia O'Neill, Xilin Long, Tania E. Webb, Eric A. Barnard, Edward Lakatta, Marvin O. Boluyt

Research output: Contribution to journalArticle

Abstract

Objectives: The aims of this study were to determine (1) whether neonatal rat cardiac fibroblasts (CAFB) express P2Y receptors; (2) whether CAFB respond to extracellular ATP by inducing expression of c-fos mRNA; and (3) whether extracellular ATP modulates norepinephrine (NE)-stimulated cell growth in CAFB. Methods: Expression of P2Y1 and P2Y2 receptors and induction of c-fos were examined by Northern blot analysis. CAFB growth was assessed by measuring [3H]thymidine incorporation and DNA content. P2Y receptor pharmacology was studied using various ATP analogues. Results: Northern blot analysis of polyA enriched RNA confirmed that at least 2 subtypes of P2Y receptors (P2Y1 and P2Y2) are expressed in cultured CAFB. Extracellular ATP induced the expression of c-fos mRNA through a pathway that was sensitive to inhibitors of protein kinase C (PKC), but not to inhibitors of intracellular Ca2+ signaling. Extracellular ATP inhibited the NE- stimulated increases in DNA content and in [3H]thymidine incorporation into DNA. Whereas the potency order for stimulation of c-fos expression was ATP = UTP > ADP > adenosine, the potency order to inhibit the NE-induced increase of [3H]thymidine incorporation into DNA was ATP > ADP > UTP > adenosine. Conclusions: These data demonstrate that CAFB express both P2Y1 and P2Y2 receptor mRNA and that CAFB respond to P2Y receptor stimulation by induction of c-fos and inhibition of DNA synthesis. These findings suggest that the effects of ATP on [3H]thymidine incorporation into DNA and on expression of c-fos mRNA are exerted via distinct P2Y receptor subtypes.

Original languageEnglish (US)
Pages (from-to)718-728
Number of pages11
JournalCardiovascular Research
Volume37
Issue number3
DOIs
StatePublished - Mar 1998
Externally publishedYes

Fingerprint

fos Genes
Fibroblasts
Adenosine Triphosphate
Gene Expression
Purinergic P2Y2 Receptors
Purinergic P2Y1 Receptors
DNA
Thymidine
Norepinephrine
Messenger RNA
Uridine Triphosphate
Northern Blotting
Adenosine
Adenosine Diphosphate
Growth
Protein Kinase C
Pharmacology
RNA

Keywords

  • C-fos
  • Cell proliferation
  • P2Y receptor
  • Rat cardiac fibroblast

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Stimulation of P2Y receptors activates c-fos gene expression and inhibits DNA synthesis in cultured cardiac fibroblasts. / Zheng, Jing Sheng; O'Neill, Lydia; Long, Xilin; Webb, Tania E.; Barnard, Eric A.; Lakatta, Edward; Boluyt, Marvin O.

In: Cardiovascular Research, Vol. 37, No. 3, 03.1998, p. 718-728.

Research output: Contribution to journalArticle

Zheng, Jing Sheng ; O'Neill, Lydia ; Long, Xilin ; Webb, Tania E. ; Barnard, Eric A. ; Lakatta, Edward ; Boluyt, Marvin O. / Stimulation of P2Y receptors activates c-fos gene expression and inhibits DNA synthesis in cultured cardiac fibroblasts. In: Cardiovascular Research. 1998 ; Vol. 37, No. 3. pp. 718-728.
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AU - Zheng, Jing Sheng

AU - O'Neill, Lydia

AU - Long, Xilin

AU - Webb, Tania E.

AU - Barnard, Eric A.

AU - Lakatta, Edward

AU - Boluyt, Marvin O.

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AB - Objectives: The aims of this study were to determine (1) whether neonatal rat cardiac fibroblasts (CAFB) express P2Y receptors; (2) whether CAFB respond to extracellular ATP by inducing expression of c-fos mRNA; and (3) whether extracellular ATP modulates norepinephrine (NE)-stimulated cell growth in CAFB. Methods: Expression of P2Y1 and P2Y2 receptors and induction of c-fos were examined by Northern blot analysis. CAFB growth was assessed by measuring [3H]thymidine incorporation and DNA content. P2Y receptor pharmacology was studied using various ATP analogues. Results: Northern blot analysis of polyA enriched RNA confirmed that at least 2 subtypes of P2Y receptors (P2Y1 and P2Y2) are expressed in cultured CAFB. Extracellular ATP induced the expression of c-fos mRNA through a pathway that was sensitive to inhibitors of protein kinase C (PKC), but not to inhibitors of intracellular Ca2+ signaling. Extracellular ATP inhibited the NE- stimulated increases in DNA content and in [3H]thymidine incorporation into DNA. Whereas the potency order for stimulation of c-fos expression was ATP = UTP > ADP > adenosine, the potency order to inhibit the NE-induced increase of [3H]thymidine incorporation into DNA was ATP > ADP > UTP > adenosine. Conclusions: These data demonstrate that CAFB express both P2Y1 and P2Y2 receptor mRNA and that CAFB respond to P2Y receptor stimulation by induction of c-fos and inhibition of DNA synthesis. These findings suggest that the effects of ATP on [3H]thymidine incorporation into DNA and on expression of c-fos mRNA are exerted via distinct P2Y receptor subtypes.

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KW - Cell proliferation

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