title: P2X1 ion channels promote neutrophil chemotaxis through Rho kinase activation.
abstract: ATP, released at the leading edge of migrating neutrophils, amplifies chemotactic signals. The aim of our study was to investigate whether neutrophils express ATP-gated P2X(1) ion channels and whether these channels could play a role in chemotaxis. Whole-cell patch clamp experiments showed rapidly desensitizing currents in both human and mouse neutrophils stimulated with P2X(1) agonists, alphabeta-methylene ATP (alphabetaMeATP) and betagammaMeATP. These currents were strongly impaired or absent in neutrophils from P2X(1)(-/-) mice. In Boyden chamber assays, alphabetaMeATP provoked chemokinesis and enhanced formylated peptide- and IL-8-induced chemotaxis of human neutrophils. This agonist similarly increased W-peptide-induced chemotaxis of wild-type mouse neutrophils, whereas it had no effect on P2X(1)(-/-) neutrophils. In human as in mouse neutrophils, alphabetaMeATP selectively activated the small RhoGTPase RhoA that caused reversible myosin L chain phosphorylation. Moreover, the alphabetaMeATP-elicited neutrophil movements were prevented by the two Rho kinase inhibitors, Y27632 and H1152. In a gradient of W-peptide, P2X(1)(-/-) neutrophils migrated with reduced speed and displayed impaired trailing edge retraction. Finally, neutrophil recruitment in mouse peritoneum upon Escherichia coli injection was enhanced in wild-type mice treated with alphabetaMeATP, whereas it was significantly impaired in the P2X(1)(-/-) mice. Thus, activation of P2X(1) ion channels by ATP promotes neutrophil chemotaxis, a process involving Rho kinase-dependent actomyosin-mediated contraction at the cell rear. These ion channels may therefore play a significant role in host defense and inflammation.
authors.affiliation: GIGA-Research Human Genetics Unit, University of Liège, Liège, Belgium.
authors.forename: Christelle
authors.initials: C
authors.lastname: Lecut
authors.forename: Kim
authors.initials: K
authors.lastname: Frederix
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Johnson
authors.forename: Christophe
authors.initials: C
authors.lastname: Deroanne
authors.forename: Marc
authors.initials: M
authors.lastname: Thiry
authors.forename: Céline
authors.initials: C
authors.lastname: Faccinetto
authors.forename: Raphaël
authors.initials: R
authors.lastname: Marée
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Evans
authors.forename: Paul G A
authors.initials: PG
authors.lastname: Volders
authors.forename: Vincent
authors.initials: V
authors.lastname: Bours
authors.forename: Cécile
authors.initials: C
authors.lastname: Oury
mesh_terms: D000205:Actomyosin / Q000502:physiology; D000255:Adenosine Triphosphate / Q000502:physiology; D000818:Animals; D002634:Chemotaxis, Leukocyte / Q000235:genetics / Q000276:immunology*; D004789:Enzyme Activation / Q000276:immunology; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D020556:Neutrophil Infiltration / Q000235:genetics / Q000276:immunology*; D009504:Neutrophils / Q000166:cytology / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D010529:Peritoneal Cavity / Q000166:cytology; D018048:Receptors, Purinergic P2 / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D058469:Receptors, Purinergic P2X; D020742:rhoA GTP-Binding Protein / Q000378:metabolism* / Q000502:physiology
chemical_list: D018048:Receptors, Purinergic P2; D058469:Receptors, Purinergic P2X; D000255:Adenosine Triphosphate; D000205:Actomyosin; D020742:rhoA GTP-Binding Protein