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http://dx.doi.org/10.14348/molcells.2015.2357

The Signaling Mechanism of Contraction Induced by ATP and UTP in Feline Esophageal Smooth Muscle Cells  

Kwon, Tae Hoon (Department of Pharmacology, College of Pharmacy, Chung-Ang University)
Jung, Hyunwoo (Department of Pharmacology, College of Pharmacy, Chung-Ang University)
Cho, Eun Jeong (Department of Pharmacology, College of Pharmacy, Chung-Ang University)
Jeong, Ji Hoon (Department of Pharmacology, College of Medicine, Chung-Ang University)
Sohn, Uy Dong (Department of Pharmacology, College of Pharmacy, Chung-Ang University)
Publication Information
Molecules and Cells / v.38, no.7, 2015 , pp. 616-623 More about this Journal
Abstract
P2 receptors are membrane-bound receptors for extracellular nucleotides such as ATP and UTP. P2 receptors have been classified as ligand-gated ion channels or P2X receptors and G protein-coupled P2Y receptors. Recently, purinergic signaling has begun to attract attention as a potential therapeutic target for a variety of diseases especially associated with gastroenterology. This study determined the ATP and UTP-induced receptor signaling mechanism in feline esophageal contraction. Contraction of dispersed feline esophageal smooth muscle cells was measured by scanning micrometry. Phosphorylation of $MLC_{20}$ was determined by western blot analysis. ATP and UTP elicited maximum esophageal contraction at 30 s and $10{\mu}M$ concentration. Contraction of dispersed cells treated with $10{\mu}M$ ATP was inhibited by nifedipine. However, contraction induced by $0.1{\mu}M$ ATP, $0.1{\mu}M$ UTP and $10{\mu}M$ UTP was decreased by U73122, chelerythrine, ML-9, PTX and $GDP{\beta}S$. Contraction induced by $0.1{\mu}M$ ATP and UTP was inhibited by $G{\alpha}i_3$ or $G{\alpha}q$ antibodies and by $PLC{\beta}_1$ or $PLC{\beta}_3$ antibodies. Phosphorylated $MLC_{20}$ was increased by ATP and UTP treatment. In conclusion, esophageal contraction induced by ATP and UTP was preferentially mediated by P2Y receptors coupled to $G{\alpha}i_3$ and $G{\alpha}q$ proteins, which activate $PLC{\beta}_1$ and $PLC{\beta}_3$. Subsequently, increased intracellular $Ca^{2+}$ and activated PKC triggered stimulation of MLC kinase and inhibition of MLC phosphatase. Finally, increased $pMLC_{20}$ generated esophageal contraction.
Keywords
ATP; contraction; Esophagus; P2Y receptor; UTP;
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1 Fredholm, B.B., Abbracchio, M.P., Burnstock, G., Daly, J.W., Harden, T.K., Jacobson, K.A., Leff, P., and Williams, M. (1994). Nomenclature and classification of purinoceptors. Pharmacol. Rev. 46, 143-156.
2 Fredholm, B.B., Abbracchio, M.P., Burnstock, G., Dubyak, G.R., Harden, T.K., Jacobson, K.A., Schwabe, U., and Williams, M. (1997). Towards a revised nomenclature for P1 and P2 receptors. Trends Pharmacol. Sci.18, 79-82.
3 Gilman, A.G. (1987). G proteins: transducers of receptor-generated signals. Annu. Rev. Biochem. 56, 615-649.   DOI   ScienceOn
4 Haeberle, J.R., Hathaway, D.R., and DePaoli-Roach, A.A. (1985). Dephosphorylation of myosin by the catalytic subunit of a type-2 phosphatase produces relaxation of chemically skinned uterine smooth muscle. J. Biol. Chem. 260, 9965-9968.
5 Harden, T.K., Boyer, J.L., and Nicholas, R.A. (1995). P2-purinergic receptors: subtype-associated signaling responses and structure. Annu. Rev. Pharmacol.Toxicol. 35, 541-579.   DOI   ScienceOn
6 Horowitz, A., Clement-Chomienne, O., Walsh, M.P., and Morgan, K.G. (1996). Epsilon-isoenzyme of protein kinase C induces a Ca(2+)-independent contraction in vascular smooth muscle. Am. J. Physiol. 271, C589-594.   DOI
7 Huang, J., Zhou, H., Mahavadi, S., Sriwai, W., Lyall, V., and Murthy, K.S. (2005). Signaling pathways mediating gastrointestinal smooth muscle contraction and MLC20 phosphorylation by motilin receptors. Am. J. Physiol. Gastrointest. Liver Physiol. 288, G23-31.   DOI   ScienceOn
8 Ijzer, J., Kisjes, J.R., Penning, L.C., Rothuizen, J., and van, den, Ingh, T.S. (2009). The progenitor cell compartment in the feline liver: an (immuno)histochemical investigation. Vet. Pathol. 46, 614-621.   DOI   ScienceOn
9 Ikebe, M., Hartshorne, D.J., and Elzinga, M. (1987). Phosphorylation of the 20,000-dalton light chain of smooth muscle myosin by the calcium-activated, phospholipid-dependent protein kinase. Phosphorylation sites and effects of phosphorylation. J. Biol. Chem. 262, 9569-9573.
10 Jiang, L.H., Kim, M., Spelta, V., Bo, X., Surprenant, A., and North, R.A. (2003). Subunit arrangement in P2X receptors. J. Neurosci. 23, 8903-8910.
11 Lazarowski, E.R., and Harden, T.K. (1994). Identification of a uridine nucleotide-selective G-protein-linked receptor that activates phospholipase-C. J. Biol. Chem. 269, 11830-11836.
12 Lechleiter, J., Hellmiss, R., Duerson, K., Ennulat, D., David, N., Clapham, D., and Peralta, E. (1990). Distinct sequence elements control the specificity of G protein activation by muscarinic acetylcholine receptor subtypes. EMBO J. 13, 4381-4390.
13 Lee, H.Y., Bardini, M., and Burnstock, G. (2000). P2X receptor immunoreactivity in the male genital organs of the rat. Cell Tissue Res. 300, 321-330.   DOI
14 Lefebvre, R.A. (1993). Non-adrenergic non-cholinergic neurotransmission in the proximal stomach. Gen. Pharmac. 24, 257-266.   DOI   ScienceOn
15 Lundberg, J.M. (1996). Pharmacology of cotransmission in the autonomic nervous system: integrative aspects on amines, neuropeptides, adenosine triphosphate, amino acids and nitric oxide. Pharmacol. Rev. 48, 113-178.
16 Matsuda, N.M., and Miller, S.M. (2010). Non-adrenergic noncholinergic inhibition of gastrointestinal smooth muscle and its intracellular mechanism(s). Fundam. Clin. Pharmacol. 24, 261-268.
17 Murthy, K.S., McHenry, L., Grider, J.R., and Makhlouf, G.M. (1995). Adenosine A1 and A2b receptors coupled to distinct interactive signaling pathways in intestinal muscle cells. J. Pharmacol. Exp. Ther. 274, 300-306.
18 Nam, Y.S., Suh, J.S., Song, H.J., and Sohn, U.D. (2013). Signaling pathway of lysophosphatidic Acid-induced contraction in feline esophageal smooth muscle cells. Korean J. Physiol. Pharmacol. 17, 139-147.   DOI   ScienceOn
19 Murthy, K.S., and Makhlouf, G.M. (1998). Coexpression of ligandgated P2X and G protein-coupled P2Y receptors in smooth muscle. Preferential activation of P2Y receptors coupled to phospholipase C (PLC)-beta1 via Galphaq/11 and to PLC-beta3 via Gbetagammai3. J. Biol. Chem. 273, 4695-4704.   DOI   ScienceOn
20 Murthy, K.S., Zhou, H., Grider, J.R., Brautigan, D.L., Eto, M., and Makhlouf, G.M. (2003). Differential signalling by muscarinic receptors in smooth muscle: m2-mediated inactivation of myosin light chain kinase via Gi3, Cdc42/Rac1 and p21-activated kinase 1 pathway, and m3-mediated MLC20 (20 kDa regulatory light chain of myosin II) phosphorylation via Rho-associated kinase /myosin phosphatase targeting subunit 1 and protein kinase C/CPI-17 pathway. Biochem. J. 374, 145-155.   DOI
21 Puetz, S., Lubomirov, L.T., and Pfitzer, G. (2009). Regulation of Smooth Muscle Contraction by Small GTPases. Physiology 24, 342-356.   DOI   ScienceOn
22 Ralevic, V., and Burnstock, G. (1998). Receptors for purines and pyrimidines. Pharmacol. Rev. 50, 413-492.
23 Shim, J.O., Shin, C.Y., Lee, T.S., Yang, S.J., An, J.Y., Song, H.J., Kim, T.H., Huh, I.H., and Sohn, U.D. (2002). Signal transduction mechanism via adenosine A1 receptor in the cat esophageal smooth muscle cells. Cell. Signal. 14, 365-372.   DOI   ScienceOn
24 Sohn, U.D., Harnett, K.M., De Petris, G., Behar, J., and Biancani, P. (1993). Distinct muscarinic receptors, G proteins and phospholipases in esophageal and lower esophageal sphincter circular muscle. J. Pharmacol. Exp. Ther. 267, 1205-1214.
25 Somlyo, A.P., and Somlyo, A.V. (1994). Signal transduction and regulation in smooth muscle. Nature 372, 231-236.   DOI   ScienceOn
26 Sohn, U.D., Han, B., Tashjian, A.H., Jr., Behar, J., and Biancani, P. (1995a). Agonist-independent, muscle-type-specific signal-transduction pathways in cat esophageal and lower eso-phageal sphincter circular smooth-muscle. J. Pharmacol. Exp. Ther. 273, 482-491.
27 Sohn, U.D., Han, B., Tashjian, A.H., Jr., Behar, J., and Biancani, P. (1995b). Agonist-independent, muscle-type-specific signal transduction pathways in cat esophageal and lower esophageal sphincter circular smooth muscle. J. Pharmacol Exp. Ther. 273, 482-491.
28 Sohn, U.D., Harnett, K.M., Cao, W., Rich, H., Kim, N., Behar, J., and Biancani, P. (1997). Acute experimental esophagitis activates a second signal transduction pathway in cat smooth muscle from the lower esophageal sphincter. J. Pharmacol. Exp. Ther. 283, 1293-1304.
29 Surprenant, A. and North, R.A. (2009) Signaling at Purinergic P2X Receptors. Annu. Rev. Physiol. 71, 333-359.   DOI   ScienceOn
30 van der Weyden, L., Conigrave, A.D., and Morris, M.B. (2000). Signal transduction and white cell maturation via extracellular ATP and the P2Y11 receptor. Immunol. Cell Biol. 78, 369-374.   DOI   ScienceOn
31 Vantrappen, G., Janssens, J., Coremans, G. and Jian, R. (1986). Gastrointestinal motility disorders. Dig. Dis. Sci. 31, 5S-25S.   DOI
32 Webb, R.C. (2003). Smooth muscle contraction and relaxation. Adv. Physiol. Educ. 27, 201-206.   DOI
33 Yang, S.J., An, J.Y., Shim, J.O., Park, C.H., Huh, I.H., and Sohn, U.D. (2000). The mechanism of contraction by 2-chloroadenosine in cat detrusor muscle cells. J. Urol. 163, 652-658.   DOI   ScienceOn
34 Billah, M.M., and Anthes, J.C. (1990). The regulation and cellular functions of phosphatidylcholine hydrolysis. Biochem J. 269, 281-291.   DOI
35 Yiangou, Y., Facer, P., Baecker, P.A., Ford, A.P., Knowles, C.H., Chan, C.L.H., Williams, N.S., and Anand, P. (2001). ATP-gated ion channel P2X(3) is increased in human inflammatory bowel disease. Neurogastroenterol. Motil. 13, 365-369.   DOI   ScienceOn
36 Abbracchio, M.P., Burnstock, G., Boeynaems, J.M., Barnard, E.A., Boyer, J.L., Kennedy, C., Knight, G.E., Fumagalli, M., Gachet, C., Jacobson, K.A., et al. (2006). International union of pharmacology LVIII: Update on the P2Y G protein-coupled nucleotide receptors: From molecular mechanisms and pathophysiology to therapy. Pharmacol. Rev. 58, 281-341.   DOI   ScienceOn
37 Akbar, G.K., Dasari, V.R., Webb, T.E., Ayyanathan, K., Pillarisetti, K., Sandhu, A.K., Athwal, R.S., Daniel, J.L., Ashby, B., Barnard, E.A., et al. (1996). Molecular cloning of a novel P2 purinoceptor from human erythroleukemia cells. J. Biol. Chem. 271, 18363-18367.   DOI   ScienceOn
38 Bialojan, C., Ruegg, J.C., and DiSalvo, J. (1987). A myosin phosphatase modulates contractility in skinned smooth muscle. Pflugers Arch. 410, 304-312.   DOI
39 Biancani, P., Hillemeier, C., Bitar, K.N., and Makhlouf, G.M. (1987). Contraction mediated by $Ca^{2+}$ influx in esophageal muscle and by $Ca^{2+}$ release in the LES. Am. J. Physiol. 253, G760-766.
40 Bitar, K.N., Bradford, P.G., Putney, J.W., Jr., and Makhlouf, G.M. (1986). Stoichiometry of contraction and $Ca^{2+}$ mobilization by inositol 1,4,5-trisphosphate in isolated gastric smooth muscle cells. J. Biol. Chem. 261, 16591-16596.
41 Burnstock, G. (2006). Pathophysiology and therapeutic potential of purinergic signaling. Pharmacol. Rev. 58, 58-86.   DOI   ScienceOn
42 Chang, K., Hanaoka, K., Kumada, M., and Takuwa, Y. (1995). Molecular cloning and functional analysis of a novel P2 nucleotide receptor. J. Biol. Chem. 270, 26152-26158.   DOI   ScienceOn
43 Burnstock, G. (2008). Purinergic receptors as future targets for treatment of functional GI disorders. Gut 57, 1193-1194.   DOI   ScienceOn
44 Burnstock, G., Campbell, G., Satchell, D., and Smythe, A. (1997). Evidence that adenosine triphosphate or a related nucleotide is the transmitter substance released by nonadrenergic inhibitory nerves in the gut (Reprinted from Brit J Pharmacol, vol 40, pp 668-688, 1970). Br. J. Pharmacol.120, 337-357.   DOI   ScienceOn
45 Cao, W., Chen, Q., Sohn, U.D., Kim, N., Kirber, M.T., Harnett, K.M., Behar, J., and Biancani, P. (2001). $Ca^{2+}$-induced contraction of cat esophageal circular smooth muscle cells. Am. J. Physiol. Cell. Physiol. 280, C980-992.   DOI
46 Cho, Y.R., Jang, H.S., Kim, W., Park, S.Y., and Sohn, U.D. (2010). P2X and P2Y receptors mediate contraction induced by electrical field stimulation in feline esophageal smooth muscle. Korean J. Physiol. Pharmacol.14, 311-316.   DOI   ScienceOn
47 Cowen, D.S., Sanders, M., and Dubyak, G. (1990). P2-purinergic receptors activate a guanine nucleotide-dependent phospholipase C in membranes from HL-60 cells. Biochim. Biophys. Acta 1053, 195-203.   DOI   ScienceOn
48 Dubyak, G.R., and el-Moatassim, C. (1993). Signal transduction via P2-purinergic receptors for extracellular ATP and other nucleotides. Am. J. Physiol. 265, C577-606.   DOI
49 Fabiato, A., and Fabiato, F. (1979). Calculator programs for computing the composition of the solutions containing multiple metals and ligands used for experiments in skinned muscle cells. J. Physiol. (Paris) 75, 463-505.