Identification of contractile P2Y1, P2Y6 and P2Y12 receptors in rat intrapulmonary artery using selective ligands

Callum Mitchell, Nawazish-I-Husain Syed, Asrin Tengah, Alison Gurney, Charles Kennedy

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

ATP and UDP constrict rat intrapulmonary arteries, but which receptors mediate these actions is unclear. Here we used selective agonists and antagonists, along with measurements of P2Y receptor expression, to characterise the receptor subtypes involved. Isometric tension was recorded from endothelium-denuded rat intrapulmonary artery rings (i.d. 200-500 μm) mounted on a wire myograph. Expression of P2Y receptor subtype expression was determined using RT-PCR with receptor-specific oligonucleotide primers. The selective P2Y(1) agonist MRS2365 induced small, concentration-dependent contractions that were inhibited by the P2Y(1) antagonist MRS2179. Contractions evoked by ATP were unaffected by MRS2179, but inhibited by about one third by the P2Y(12) antagonist AR-C69931MX. Combined blockade of P2X1 and P2Y(12) receptors virtually abolished the response to ATP. ADP also evoked contractions that were abolished by AR-C69931MX. The selective P2Y(6) receptor agonist, PSB 0474, evoked concentration-dependent contractions and was approximately 3-times more potent than UDP, but the P2Y(14) agonist, UDP-glucose, had no effect. Contractions evoked by UDP were inhibited by the P2Y(6) receptor antagonist, MRS2578, but not the CysLT(1) antagonist, MK571. Higher concentrations of MRS2578 inhibited contractions to KCl and so were not studied further. mRNA for P2Y(1), P2Y(6) and P2Y(12) receptors was identified, but this could not be correlated with protein expression. Our working model is that P2Y(12) and P2X1 receptors are present in rat intrapulmonary arteries and together mediate ATP-induced vasoconstriction. Contractile P2Y(6), but not P2Y(14) or CysLT(1) receptors are also present and are a major site through which UDP evokes constriction.
LanguageEnglish
Pages755-762
Number of pages8
JournalJournal of Pharmacology and Experimental Therapeutics
Volume343
Issue number3
Early online date18 Sep 2012
DOIs
Publication statusPublished - Dec 2012

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Uridine Diphosphate
Arteries
Adenosine Triphosphate
Ligands
Purinergic P2X1 Receptors
Uridine Diphosphate Glucose
DNA Primers
Vasoconstriction
Constriction
Adenosine Diphosphate
Endothelium
Polymerase Chain Reaction
Messenger RNA
purinoceptor P2Y6
Proteins
N(6)-methyl-2'-deoxyadenosine 3',5'-diphosphate
N,N''-1,4-butanediylbis(N'-(3-isothiocyanatophenyl))thiourea
cangrelor

Keywords

  • P2X receptors
  • P2Y receptors
  • pulmonary pharmacology
  • purinergic receptors
  • purines
  • smooth muscle

Cite this

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title = "Identification of contractile P2Y1, P2Y6 and P2Y12 receptors in rat intrapulmonary artery using selective ligands",
abstract = "ATP and UDP constrict rat intrapulmonary arteries, but which receptors mediate these actions is unclear. Here we used selective agonists and antagonists, along with measurements of P2Y receptor expression, to characterise the receptor subtypes involved. Isometric tension was recorded from endothelium-denuded rat intrapulmonary artery rings (i.d. 200-500 μm) mounted on a wire myograph. Expression of P2Y receptor subtype expression was determined using RT-PCR with receptor-specific oligonucleotide primers. The selective P2Y(1) agonist MRS2365 induced small, concentration-dependent contractions that were inhibited by the P2Y(1) antagonist MRS2179. Contractions evoked by ATP were unaffected by MRS2179, but inhibited by about one third by the P2Y(12) antagonist AR-C69931MX. Combined blockade of P2X1 and P2Y(12) receptors virtually abolished the response to ATP. ADP also evoked contractions that were abolished by AR-C69931MX. The selective P2Y(6) receptor agonist, PSB 0474, evoked concentration-dependent contractions and was approximately 3-times more potent than UDP, but the P2Y(14) agonist, UDP-glucose, had no effect. Contractions evoked by UDP were inhibited by the P2Y(6) receptor antagonist, MRS2578, but not the CysLT(1) antagonist, MK571. Higher concentrations of MRS2578 inhibited contractions to KCl and so were not studied further. mRNA for P2Y(1), P2Y(6) and P2Y(12) receptors was identified, but this could not be correlated with protein expression. Our working model is that P2Y(12) and P2X1 receptors are present in rat intrapulmonary arteries and together mediate ATP-induced vasoconstriction. Contractile P2Y(6), but not P2Y(14) or CysLT(1) receptors are also present and are a major site through which UDP evokes constriction.",
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Identification of contractile P2Y1, P2Y6 and P2Y12 receptors in rat intrapulmonary artery using selective ligands. / Mitchell, Callum; Syed, Nawazish-I-Husain; Tengah, Asrin; Gurney, Alison; Kennedy, Charles.

In: Journal of Pharmacology and Experimental Therapeutics, Vol. 343, No. 3, 12.2012, p. 755-762.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Identification of contractile P2Y1, P2Y6 and P2Y12 receptors in rat intrapulmonary artery using selective ligands

AU - Mitchell, Callum

AU - Syed, Nawazish-I-Husain

AU - Tengah, Asrin

AU - Gurney, Alison

AU - Kennedy, Charles

PY - 2012/12

Y1 - 2012/12

N2 - ATP and UDP constrict rat intrapulmonary arteries, but which receptors mediate these actions is unclear. Here we used selective agonists and antagonists, along with measurements of P2Y receptor expression, to characterise the receptor subtypes involved. Isometric tension was recorded from endothelium-denuded rat intrapulmonary artery rings (i.d. 200-500 μm) mounted on a wire myograph. Expression of P2Y receptor subtype expression was determined using RT-PCR with receptor-specific oligonucleotide primers. The selective P2Y(1) agonist MRS2365 induced small, concentration-dependent contractions that were inhibited by the P2Y(1) antagonist MRS2179. Contractions evoked by ATP were unaffected by MRS2179, but inhibited by about one third by the P2Y(12) antagonist AR-C69931MX. Combined blockade of P2X1 and P2Y(12) receptors virtually abolished the response to ATP. ADP also evoked contractions that were abolished by AR-C69931MX. The selective P2Y(6) receptor agonist, PSB 0474, evoked concentration-dependent contractions and was approximately 3-times more potent than UDP, but the P2Y(14) agonist, UDP-glucose, had no effect. Contractions evoked by UDP were inhibited by the P2Y(6) receptor antagonist, MRS2578, but not the CysLT(1) antagonist, MK571. Higher concentrations of MRS2578 inhibited contractions to KCl and so were not studied further. mRNA for P2Y(1), P2Y(6) and P2Y(12) receptors was identified, but this could not be correlated with protein expression. Our working model is that P2Y(12) and P2X1 receptors are present in rat intrapulmonary arteries and together mediate ATP-induced vasoconstriction. Contractile P2Y(6), but not P2Y(14) or CysLT(1) receptors are also present and are a major site through which UDP evokes constriction.

AB - ATP and UDP constrict rat intrapulmonary arteries, but which receptors mediate these actions is unclear. Here we used selective agonists and antagonists, along with measurements of P2Y receptor expression, to characterise the receptor subtypes involved. Isometric tension was recorded from endothelium-denuded rat intrapulmonary artery rings (i.d. 200-500 μm) mounted on a wire myograph. Expression of P2Y receptor subtype expression was determined using RT-PCR with receptor-specific oligonucleotide primers. The selective P2Y(1) agonist MRS2365 induced small, concentration-dependent contractions that were inhibited by the P2Y(1) antagonist MRS2179. Contractions evoked by ATP were unaffected by MRS2179, but inhibited by about one third by the P2Y(12) antagonist AR-C69931MX. Combined blockade of P2X1 and P2Y(12) receptors virtually abolished the response to ATP. ADP also evoked contractions that were abolished by AR-C69931MX. The selective P2Y(6) receptor agonist, PSB 0474, evoked concentration-dependent contractions and was approximately 3-times more potent than UDP, but the P2Y(14) agonist, UDP-glucose, had no effect. Contractions evoked by UDP were inhibited by the P2Y(6) receptor antagonist, MRS2578, but not the CysLT(1) antagonist, MK571. Higher concentrations of MRS2578 inhibited contractions to KCl and so were not studied further. mRNA for P2Y(1), P2Y(6) and P2Y(12) receptors was identified, but this could not be correlated with protein expression. Our working model is that P2Y(12) and P2X1 receptors are present in rat intrapulmonary arteries and together mediate ATP-induced vasoconstriction. Contractile P2Y(6), but not P2Y(14) or CysLT(1) receptors are also present and are a major site through which UDP evokes constriction.

KW - P2X receptors

KW - P2Y receptors

KW - pulmonary pharmacology

KW - purinergic receptors

KW - purines

KW - smooth muscle

U2 - 10.1124/jpet.112.198051

DO - 10.1124/jpet.112.198051

M3 - Article

VL - 343

SP - 755

EP - 762

JO - Journal of Pharmacology and Experimental Therapeutics

T2 - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

SN - 0022-3565

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