Differential Regulation and Relocalization of the Platelet P2Y Receptors after Activation: A Way to Avoid Loss of Hemostatic Properties?

Baurand, Anthony; Eckly, Anita; Hechler, Béatrice; Kauffenstein, Gilles; Galzi, Jean‐Luc; Cazenave, Jean‐Pierre; Léon, Catherine; Gachet, Christian

doi:10.1124/mol.104.004846

Cited by 70 publications

(79 citation statements)

References 36 publications

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“…As with other GPCRs, the responsiveness of P2Y 1 and P2Y 12 receptors is tightly regulated. Recently, we showed that both P2Y 1 and P2Y 12 receptor responses desensitize in human platelets [4], in agreement with the observed desensitization of platelet responses following prolonged exposure to ADP [5][6][7]. Mechanisms underlying desensitization are complex and can involve phosphorylation of the receptor, uncoupling from G proteins, internalization, and ultimately intracellular downregulation [3,[8][9][10][11][12].…”

Section: Introductionmentioning

confidence: 54%

Rapid resensitization of purinergic receptor function in human platelets

Mundell

Barton

Mayo-Martin

et al. 2008

Journal of Thrombosis and Haemostasis

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Summary. Background: Adenosine diphosphate (ADP) is a critical regulator of platelet activation, mediating its actions through two G protein-coupled receptors (GPCRs), the P2Y 1 and P2Y 12 purinergic receptors. Recently, we demonstrated that both receptors desensitize and internalize in human platelets by differential kinase-dependent mechanisms. Objectives: To demonstrate whether responses to P2Y 1 and P2Y 12 purinergic receptors resensitize in human platelets and determine the role of receptor traffic in this process. Methods: These studies were undertaken either in human platelets or in cells stably expressing epitope-tagged P2Y 1 and P2Y 12 purinergic receptor constructs. Results: In this study we show for the first time that responses to both of these receptors can rapidly resensitize following agonistdependent desensitization in human platelets. Further, we show that in human platelets or in 1321N1 cells stably expressing receptor constructs, the disruption of receptor internalization, dephosphorylation or subsequent receptor recycling is sufficient to block resensitization of purinergic receptor responses. We also show that, in platelets, internalization of both these receptors is dependent upon dynamin, and that this process is required for resensitization of responses. Conclusions: This study is therefore the first to show that both P2Y 1 and P2Y 12 receptor activities are rapidly and reversibly modulated in human platelets, and it reveals that the underlying mechanism requires receptor trafficking as an essential part of this process.

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Section: Introductionmentioning

confidence: 54%

Rapid resensitization of purinergic receptor function in human platelets

Mundell

Barton

Mayo-Martin

et al. 2008

Journal of Thrombosis and Haemostasis

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“…Both receptors (P2Y 1 and P2Y 12 ) are internalized, however, with different kinetics and through different pathways. This resulted in a greater persistence of the P2Y 12 receptor at the plasma membrane in the presence of nucleotide in the medium, while the P2Y 1 receptor remained internalized [24][25][26].…”

Section: Discussionmentioning

confidence: 99%

Regulation of death and survival in astrocytes by ADP activating P2Y1 and P2Y12 receptors

Mamedova

Gao

2006

Biochemical Pharmacology

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ADP is the endogenous agonist for both P2Y 1 and P2Y 12 receptors, which are important therapeutic targets. It was previously demonstrated that ADP and a synthetic agonist, 2-methylthioadenosine 5′-diphosphate (2MeSADP), can induce apoptosis by activating the human P2Y 1 receptor heterologously expressed in astrocytoma cells. However, it was not known whether the P2Y 12 receptor behaved similarly. We demonstrated here that, unlike with the G q -coupled P2Y 1 receptor, activation of the G i -coupled P2Y 12 receptor does not induce apoptosis. Furthermore, activation of the P2Y 12 receptor by either ADP or 2MeSADP significantly attenuates the tumor necrosis factor α (TNFα)-induced apoptosis in 1321N1 human astrocytoma cells. This protective effect was blocked by the P2Y 12 receptor antagonist 2-methylthioAMP and by inhibitors of phospholipase C (U73122) and protein kinase C (chelerythrin), but not by the P2Y 1 receptor antagonist MRS2179. Toward a greater mechanistic understanding, we showed that hP2Y 12 receptor activation by 10 nM 2MeSADP, activates Erk1/2, Akt, and JNK by phosphorylation. However, at a lower protective concentration of 100 pM 2MeSADP, activation of the hP2Y 12 receptor involves only phosphorylated Erk1/2, but not Akt or JNK. This activation is hypothesized as the major mechanism for the protective effect induced by P2Y 12 receptor activation. Apyrase did not affect the ability of TNFα to induce apoptosis in hP2Y 12 -1321N1 cells, suggesting that the endogenous nucleotides are not involved. These results may have important implications for understanding the signaling cascades that follow activation of P2Y 1 and P2Y 12 receptors and their opposing effects on cell death pathways.

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“…The molecular mechanisms of this phenomenon have been studied in detail, but consensus has not been reached, and two different views have not yet been reconciled. On the one hand, it is thought that the phenomenon of platelet refractoriness to ADP is due to selective desensitization and internalization of the P2Y 1 receptor, while the P2Y 12 receptor remains functional with the ability of ADP to induce amplification of the platelet aggregation induced by other agonists [89][90][91]. Desensitization of the P2Y 1 receptor has been shown to be dependent on receptor C-terminal phosphorylation sites, β-arrestin-2 interaction and protein kinase C (PKC) activity [92,93].…”

Section: Desensitizationmentioning

confidence: 99%

P2 receptors and platelet function

Hechler

Gachet

2011

Purinergic Signalling

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Following vessel wall injury, platelets adhere to the exposed subendothelium, become activated and release mediators such as TXA 2 and nucleotides stored at very high concentration in the so-called dense granules. Released nucleotides and other soluble agents act in a positive feedback mechanism to cause further platelet activation and amplify platelet responses induced by agents such as thrombin or collagen. Adenine nucleotides act on platelets through three distinct P2 receptors: two are G proteincoupled ADP receptors, namely the P2Y 1 and P2Y 12 receptor subtypes, while the P2X 1 receptor ligand-gated cation channel is activated by ATP. The P2Y 1 receptor initiates platelet aggregation but is not sufficient for a full platelet aggregation in response to ADP, while the P2Y 12 receptor is responsible for completion of the aggregation to ADP. The latter receptor, the molecular target of the antithrombotic drugs clopidogrel, prasugrel and ticagrelor, is responsible for most of the potentiating effects of ADP when platelets are stimulated by agents such as thrombin, collagen or immune complexes. The P2X 1 receptor is involved in platelet shape change and in activation by collagen under shear conditions. Each of these receptors is coupled to specific signal transduction pathways in response to ADP or ATP and is differentially involved in all the sequential events involved in platelet function and haemostasis. As such, they represent potential targets for antithrombotic drugs.

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Differential Regulation and Relocalization of the Platelet P2Y Receptors after Activation: A Way to Avoid Loss of Hemostatic Properties?

Cited by 70 publications

References 36 publications

Rapid resensitization of purinergic receptor function in human platelets

Rapid resensitization of purinergic receptor function in human platelets

Regulation of death and survival in astrocytes by ADP activating P2Y1 and P2Y12 receptors

P2 receptors and platelet function

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