Publication: Corneal Re-epithelialization Stimulated by Diadenosine Polyphosphates Recruits RhoA/ROCK and ERK1/2 Pathways
Loading...
Official URL
Full text at PDC
Publication Date
2008-11
Advisors (or tutors)
Editors
Journal Title
Journal ISSN
Volume Title
Publisher
The Association for Research in Vision and Ophthalmology, Inc.
Citations
Exportar
Abstract
Purpose. To investigate the role of ERK1/2 and RhoA/ROCK intracellular pathways in the modification of corneal re-epithelialization when stimulated by the diadenosine polyphosphates Ap4A and Ap3A.
Methods. In wounded confluent SIRC (Statens Seruminstitut rabbit cornea) cell monolayers and in the presence or absence of Ap4A or Ap3A 100 μM, a battery of P2 receptor antagonists and inhibitors of tyrosin kinases, MAPK, and cytoskeleton pathways (AG1478 100 μM, U0126 100 μM, Y27632 100 nM, and (−)-blebbistatin 10 μM; n = 8 each) were assayed. Also, the activation of ERK1/2 and ROCK-I was examined by Western blot assay after treatment with Ap4A and Ap3A (100 μM), with or without suramin, RB-2, U0126, and Y27632. The intracellular distribution of pERK and ROCK-I was examined in the presence of Ap4A or Ap3A (100 μM) with U0126 and Y27632 (100 nM).
Results. In the presence of Ap4A, U0126, Y27632, AG1478, and (−)-blebbistatin, reduced the migration rate compared to the effect of Ap4A alone (P < 0.0001, P < 0.001, P < 0.01, and P < 0.1 versus Ap4A, respectively). In the presence of Ap3A 100 μM, U0126 and Y27632 accelerated the migration rate when compared with the effect of Ap3A alone, whereas AG1478 and (−)-blebbistatin (P < 0.0001 versus Ap3A) slowed the migration rate. Western blot assays demonstrated that both dinucleotides activated the ERK1/2 pathway but only Ap4A activated the ROCK-I pathway. The intracellular distribution of pERK1/2 and ROCK-I reflected cross-talk between these two pathways.
Conclusions. The activation of the Ap4A/P2Y2 receptor, accelerates corneal epithelial cell migration during wound healing with the activation of MAPK and cytoskeleton pathways, whereas activation of the Ap3A/P2Y6 receptor signals only the MAPK pathway.
Description
En O.A. en la web del editor
UCM subjects
Unesco subjects
Keywords
Citation
1 BurnstockG. The past, present and future of purine nucleotides as signalling molecules. Neuropharmacology. 1997;36(9)1127–1139.
2 McLennanAG. Dinucleoside polyphosphates: friend or foe?. Pharmacol Ther. 2000;87(2–3)73–89.
3 BurnstockG, KnightGE. Cellular distribution and functions of P2 receptor subtypes in different systems. Int Rev Cytol. 2004;240:301–304.
4 RotllanP, Miras-PortugalMT. Adenosine kinase from bovine adrenal medulla. Eur J Biochem. 1985;151(2)365–371.
5 HildermanRH, ChristensenEF. P1, P4-diadenosine 5′ tetraphosphate induces nitric oxide release from bovine aortic endothelial cells. FEBS Lett. 1998;427(3)320–324.
6 ZamecnikPC, KimB, GaoMJ, TaylorG, BlackburnGM. Analogues of diadenosine 5′,5‴-P1, P4-tetraphosphate (Ap4A) as potential anti-platelet-aggregation agents. Proc Natl Acad Sci USA. 1992;89(6)2370–2373.
7 PintorJ, Diaz-HernandezM, GualixJ, Gomez-VillafuertesR, HernandoF, Miras-PortugalMT. Diadenosine polyphosphate receptors: from rat and guinea-pig brain to human nervous system. Pharmacol Ther. 2000;87(2–3)103–115. 8 BurnstockG, KennedyC. Is there a basis for distinguishing two types of P2-purinoceptor?. Gen Pharmacol. 1985;16:433–440.
9 NorthRA. Molecular physiology of P2X receptors. Physiol Rev. 2002;82:1013–1067.
10 NorthRA. P2Y purinoceptor plethora. Semin Neurosci. 1996;8:187–194.
11 RalevicV, BurnstockG. Receptors for purines and pyrimidines. Pharmacol Rev. 1998;50:413–492.
12 KhakhBS. Molecular physiology of P2X receptors and ATP signalling at synapses. Nat Rev Neurosci. 2001;2:165–174.
13 WeismanGA, GonzalezFA, ErbL, GarradRA, TurnerJT. The cloning and expression of G-coupled P2Y receptors.TurnerJT WeismanGA FedanJS eds. The P2 Nucleotide Receptors. 1998;63–79.Humana Press Inc. Totowa, NJ.
14 ChambersJK, MacDonaldLE, SarauHM, et al. A G protein-coupled receptor for UDP-glucose. J Biol Chem. 2000;275(15)10767–10771.
15 ZhangFL, LuoL, GustafsonE, et al. P2Y(13): identification and characterization of a novel Galphai-coupled ADP receptor from human and mouse. J Pharmacol Exp Ther. 2002;301(2)705–713.
16 SakK, WebbTE. A retrospective of recombinant P2Y receptor subtypes and their pharmacology. Arch Biochem Biophys. 2002;397:131–136.
17 BoeynaemsJM, CommuniD, GonzalezNS, RobayeB. Overview of the P2 receptors. Semin Thromb Hemost. 2005;31:139–149.
18 BoeynaemsJM, van GiezenH, SaviP, HerbertJM. P2Y receptor antagonists in thrombosis. Curr Opin Investig Drug. 2005;6:275–282.
19 PintorJ, PeralA, PeláezT, CarracedoG, BautistaA, HoyleCHV. Nucleotides and dinucleotides in ocular physiology: new possibilities of nucleotides as therapeutic agents in the eye. Drug Develop Res. 2003;58:1–10.
20 PintorJ, CarracedoG, AlonsoMC, BautistaA, PeralA. Presence of diadenosine polyphosphates in human tears. Pflugers Arch. 2002;443(3)432–436.
21 PintorJ, PeralA, HoyleCH, et al. Effects of diadenosine polyphosphates on tear secretion in New Zealand white rabbits. J Pharmacol Exp Ther. 2002;300(1)291–297.
22 PintorJ, BautistaA, CarracedoG, PeralA. UTP and diadenosine tetraphosphate accelerate wound healing in the rabbit cornea. Ophthalmic Physiol Opt. 2004;24(3)186–193.
23 MedieroA, PeralA, PintorJ. Dual role of diadenosine polyphosphates on corneal epithelial cell migration. Invest Ophthalmol Vis Sci. 2006;47(10)4500–4506.
24 SharmaGD, HeJ, BazanHE. p38 and ERK1/2 coordinate cellular migration and proliferation in epithelial wound healing: evidence of cross-talk activation between MAP kinase cascades. J Biol Chem. 2003;278(24)21989–21997.
25 SaikaS, OkadaY, MiyamotoT, et al. Role of p38 MAP kinase in regulation of cell migration and proliferation in healing corneal epithelium. Invest Ophthalmol Vis Sci. 2004;45(1)100–109.
26 DengM, ChenWL, TakatoriA, et al. A role for the mitogen-activated protein kinase kinase kinase 1 in epithelial wound healing. Mol Biol Cell. 2006;17(8)3446–3455.
27 WilkinsonMG, MillarJB. Control of the eukaryotic cell cycle by MAP kinase signaling pathways. FASEB J. 2000;14(14)2147–2157.
28 HallA. Ras-related GTPases and the cytoskeleton. Mol Biol Cell. 1992;3(5)475–479.
29 RidleyAJ. Signal transduction through the GTP-binding proteins Rac and Rho. J Cell Sci Suppl. 1994;18:127–131.
30 SanderEE, CollardJG. Rho-like GTPases: their role in epithelial cell-cell adhesion and invasion. Eur J Cancer. 1999;35(14)1905–1911.
31 SundarRajN, KinchingtonPR, WesselH, et al. A Rho-associated protein kinase: differentially distributed in limbal and corneal epithelia. Invest Ophthalmol Vis Sci. 1998;39(7)1266–1272. [PubMed]
32 KlepeisVE, WeingerI, KaczmarekE, Trinkaus-RandallV. P2Y receptors play a critical role in epithelial cell communication and migration. J Cell Biochem. 2004;93:1115–1133.
33 YangL, CrasonD, Trinkaus-RandallV. Cellular injury induces activation of MAPK via P2Y receptors. J Cell Biochem. 2004;91:938–950.
34 WeingerI, KlepeisVE, Trinkaus-RandallV. Tri-nucleotide receptors play a critical role in epithelial cell wound repair. Purinergic Signal. 2005;1:281–292.
35 SauzeauV, Le JeuneH, Cario-ToumaniantzC, et al. P2Y(1), P2Y(2), P2Y(4), and P2Y(6) receptors are coupled to Rho and Rho kinase activation in vascular myocytes. Am J Physiol Heart Circ Physiol. 2000;278(6)H1751–H1761.
36 CrossonCE, KlyceSD, BeuermanRW. Epithelial wound closure in the rabbit cornea: a biphasic process. Invest Ophthalmol Vis Sci. 1986;27(4)464–473.
37 ValsterA, TranNL, NakadaM, BerensME, ChanAY, SymonsM. Cell migration and invasion assays. Methods. 2005;37:208–215.
38 AshleyN, HarrisD, PoultonJ. Detection of mitochondrial DNA depletion in living human cells using PicoGreen staining. Exp Cell Res. 2005;303(2)432–446.
39 DesaiLP, AryalAM, CeacareanuB, HassidA, WatersCM. RhoA and Rac1 are both required for efficient wound closure of airway epithelial cells. Am J Physiol Lung Cell Mol Physiol. 2005;287:L1134–L1144.
40 LeeJG, KayEP. FGF-2-induced wound healing in corneal endothelial cells requires Cdc42 activation and rho inactivation through the phosphatidylinositol 3-kinase pathway. Invest Ophthalmol Vis Sci. 2006;47(4)1376–1386.
41 BrunetA, RouxD, LenormandP, DowdS, KeyseS, PouysségurJ. Nuclear translocation of p42/p44 mitogen-activated protein kinase is required for growth factor-induced gene expression and cell cycle entry. EMBO J. 1999;18(3)664–674.
42 Guzman-AranguezA, CrookeA, PeralA, HoyleCHV, PintorJ. Dinucleoside polyphosphates in the eye: from physiology to therapeutics. Prog Retin Eye Res. 2007;26(6)674–687.
43 AhmadS, AhmadA, GhoshM, LeslieCC, WhiteCW. Extracellular ATP-mediated signaling for survival in hyperoxia-induced oxidative stress. J Biol Chem. 2004;279:16317–16325.
44 DelicadoEG, JimenezAI, CarrasqueroLMG, CastroE, Miras-PortugalMT. Cross-talk among epidermal growth factor, Ap(5)A, and nucleotide receptors causing enhanced ATP Ca(2+) signaling involves extracellular kinase activation in cerebellar astrocytes. J Neurosci Res. 2005;81:789–796.
45 DouilletCD, RobinsonWP, 3rd, MilanoPM, BoucherRC, RichPB. Nucleotides induce IL-6 release from human airway epithelia via P2Y2 and p38 MAPK-dependent pathways. Am J Physiol Lung Cell Mol Physiol. 2006;291:L734–L746.
46 BessardA, CoutantA, RescanC, et al. An MLCK-dependent window in late G1 controls S phase entry of proliferating rodent hepatocytes via ERK-p70S6K pathway. Hepatology. 2006;44(1)152–163.
47 PawlakG, HelfmanDM. MEK mediates v-Src-induced disruption of the actin cytoskeleton via inactivation of the Rho-ROCK-LIM kinase pathway. J Biol Chem. 2002;277(30)26927–26933.
48 RooversK, AssoianRK. Effects of rho kinase and actin stress fibers on sustained extracellular signal-regulated kinase activity and activation of G(1) phase cyclin-dependent kinases. Mol Cell Biol. 2003;23(12)4283–4294.
49 SuzukiK, SaitoJ, YanaiR, et al. Cell-matrix and cell-cell interactions during corneal epithelial wound healing. Prog Retin Eye Res. 2003;22(2)113–133.
50 WillardFS, CrouchMF. MEK, ERK, and p90RSK are present on mitotic tubulin in Swiss 3T3 cells: a role for the MAP kinase pathway in regulating mitotic exit. Cell Signal. 2001;13(9)653–664.
51 WangR, HeG, Nelman-GonzalezM, AshornCL, et al. Regulation of Cdc25C by ERK-MAP kinases during the G2/M transition. Cell. 2007;128:1119–1132.