UTP Controls Cell Surface Distribution and Vasomotor Activity of the Human P2Y2 Receptor through an Epidermal Growth Factor Receptor-transregulated Mechanism

Abstract

Extracellular nucleotides transmit signals into the cells through the P2 family of cell surface receptors. These receptors are amply expressed in human blood vessels and participate in vascular tone control; however, their signaling mechanisms remain unknown. Here we show that in smooth muscle cells of isolated human chorionic arteries, the activation of the P2Y 2 receptor (P2Y 2 R) induces not only its partition into membrane rafts but also its rapid internalization. Cholesterol depletion with methyl-␤-cyclodextrin reduced the association of the agonist-activated receptor into membrane rafts but did not affect either the UTPmediated vasoconstrictions or the vasomotor responses elicited by both serotonin and KCl. Ex vivo perfusion of human chorionic artery segments with 1-10 M UTP, a selective P2Y 2 R agonist, displaced the P2Y 2 R localization into membrane rafts within 1 min, a process preceded by the activation of both RhoA and Rac1 GTPases. AG1478, a selective and potent inhibitor of the epidermal growth factor receptor tyrosine kinase activity, not only blocked the UTP-induced vasomotor activity but also abrogated both RhoA and Rac1 activation, the P2Y 2 R association with membrane rafts, and its internalization. Altogether, these results show for the first time that the plasma membrane distribution of the P2Y 2 R is transregulated by the epidermal growth factor receptor, revealing an unsuspected functional interplay that controls both the membrane distribution and the vasomotor activity of the P2Y 2 R in intact human blood vessels.

Caveolin-1 Regulates the P2Y2 Receptor Signaling in Human 1321N1 Astrocytoma Cells

“…Use of BV-2 cells, a Cav-1 devoid microglial cell line expressing P2Y 2 Rs, substantiates this finding, because P2Y 2 R was found to reside in high density membrane fractions (Fig. 2) (67).…”

Interaction of purinergic receptors with GPCRs, ion channels, tyrosine kinase and steroid hormone receptors orchestrates cell function

“…There, the use of AG1478, a selective and potent inhibitor of the EGFR tyrosine kinase activity, not only blocked the UTP-induced vasomotor activity but also abrogated both RhoA and Rac1 activation, the P2Y 2 receptor association with membrane rafts, and its internalization. These results reveal an unsuspected functional interplay that controls both the membrane distribution and the vasomotor activity of the P2Y 2 receptor in intact human blood vessels [88].…”

Uridine adenosine tetraphosphate-induced contraction is increased in renal but not pulmonary arteries from DOCA-salt hypertensive rats

“…Various signal transduction pathways (which are associated with vasoconstriction) activated by P2Y receptor stimulation have been proposed, including MAPK, Rho A/Rho kinase, and EGF receptor (EGFR) transactivation (47,54). Moreover, several reports have demonstrated that these signal transduction pathways are altered in various animal models, including mineralocorticoid hypertension (5,18,19,59).…”