Abstract
The A2B adenosine receptor (A2BR) mediates biological responses to extracellular adenosine in a wide variety of cell types. Adenosine deaminase (ADA) can degrade adenosine and bind extracellularly to adenosine receptors. Adenosine modulates chloride secretion in gastric glands and gastric mucosa parietal cells. A close functional link between surface A2BR and ADA has been found on cells of the immune system, but whether this occurs in the gastrointestinal tract is unknown. The goal of this study was to determine whether A2BR and ADA are coexpressed at the plasma membrane of the acid-secreting gastric mucosa parietal cells. We used isolated gastric parietal cells after purification by centrifugal elutriation. The membrane fraction was obtained by sucrose gradient centrifugation. A2BR mRNA expression was analyzed by RT-PCR. The surface expression of A2BR and ADA proteins was evaluated by Western blotting, flow cytometry and confocal microscopy. Our findings demonstrate that A2BR and ADA are expressed in cell membranes isolated from gastric parietal cells. They show a high degree of colocalization that is particularly evident in the surface of contact between parietal cells. The confocal microscopy data together with flow cytometry analysis suggest a tight association between A2BR and ADA that might be specifically linked to glandular secretory function.
Similar content being viewed by others
Abbreviations
- A2BR:
-
A2B adenosine receptor
- ADA:
-
adenosine deaminase
- RT-PCR:
-
reverse transcription-polymerase chain reaction
References
Van, L. A., and Eltzschig, H. K. (2007) Role of pulmonary adenosine during hypoxia: extracellular generation, signaling and metabolism by surface adenosine deaminase/CD26, Expert Opin. Biol. Ther., 7, 1437–1447.
Fredholm, B. B., Izerman, A. P., Jacobson, K. A., Klotz, K. N., and Linden, J. (2001) International Union of Pharmacology. XXV. Nomenclature and classification of adenosine receptors, Pharmacol. Rev., 53, 527–552.
Thimm, D., Schiedel, A. C., Sherbiny, F. F., Hinz, S., Hochheiser, K., Bertarelli, D. C., Maass, A., and Muller, C. E. (2013) Ligand-specific binding and activation of the human adenosine A(2B) receptor, Biochemistry, 52, 726–740.
Kong, T., Westerman, K. A., Faigle, M., Eltzschig, H. K., and Colgan, S. P. (2006) HIF-dependent induction of adenosine A2B receptor in hypoxia, FASEB J., 20, 2242–2250.
Aherne, C. M., Kewley, E. M., and Eltzschig, H. K. (2011) The resurgence of A2B adenosine receptor signaling, Biochim. Biophys. Acta, 1808, 1329–1339.
Franco, R., Valenzuela, A., Lluis, C., and Blanco, J. (1998) Enzymatic and extraenzymatic role of ecto-adenosine deaminase in lymphocytes, Immunol. Rev., 161, 27–42.
Franco, R., Casado, V., Ciruela, F., Saura, C., Mallol, J., Canela, E. I., and Lluis, C. (1997) Cell surface adenosine deaminase: much more than an ectoenzyme, Prog. Neurobiol., 52, 283–294.
Gines, S., Marino, M., Mallol, J., Canela, E. I., Morimoto, C., Callebaut, C., Hovanessian, A., Casado, V., Lluis, C., and Franco, R. (2002) Regulation of epithelial and lymphocyte cell adhesion by adenosine deaminaseCD26 interaction, Biochem. J., 361, 203–209.
Pacheco, R., Martinez-Navio, J. M., Lejeune, M., Climent, N., Oliva, H., Gatell, J. M., Gallart, T., Mallol, J., Lluis, C., and Franco, R. (2005) CD26, adenosine deaminase, and adenosine receptors mediate costimulatory signals in the immunological synapse, Proc. Natl. Acad. Sci. USA, 102, 9583–9588.
Gracia, E., Farre, D., Cortes, A., Ferrer-Costa, C., Orozco, M., Mallol, J., Lluis, C., Canela, E. I., McCormick, P. J., Franco, R., Fanelli, F., and Casado, V. (2013) The catalytic site structural gate of adenosine deaminase allosterically modulates ligand binding to adenosine receptors, FASEB J., 27, 1048–1061.
Mirabet, M., Herrera, C., Cordero, O. J., Mallol, J., Lluis, C., and Franco, R. (1999) Expression of A2B adenosine receptors in human lymphocytes: their role in T cell activation, J. Cell. Sci., 112, 491–502.
Herrera, C., Morimoto, C., Blanco, J., Mallol, J., Arenzana, F., Lluis, C., and Franco, R. (2001) Comodulation of CXCR4 and CD26 in human lymphocytes, J. Biol. Chem., 276, 19532–19539.
Ainz, L. F., Salgado, C., Gandarias, J. M., Gomez, R., Vallejo, A., and Gil-Rodrigo, C. E. (1993) P1(A2/Ra)-purinoceptors may mediate the stimulatory effect of adenosine and adenosine analogs on acid formation in isolated rabbit parietal cells, Pharmacol. Res., 27, 319–334.
Ainz, L. F., Gil-Rodrigo, C. E., Gomez, R., Malillos, M., Requejo, D., and Gandarias, J. M. (1989) Effects of various physiologic adenine derivatives on the secretion of acid in isolated gastric glands in rabbits, Rev. Esp. Fisiol., 45, 281–286.
Gil-Rodrigo, C. E., Galdiz, B., Gandarias, J. M., Gomez, R., and Ainz, L. F. (1990) Characterization of the effects of adenosine, adenosine 5′-triphosphate and related purines on acid secretion in isolated rabbit gastric glands, Pharmacol. Res., 22, 103–113.
Colgan, S. P., Fennimore, B., and Ehrentraut, S. F. (2013) Adenosine and gastrointestinal inflammation, J. Mol. Med. (Berl.), 91, 157–164.
Wang, L., Kolachala, V., Walia, B., Balasubramanian, S., Hall, R. A., Merlin, D., and Sitaraman, S. V. (2004) Agonist-induced polarized trafficking and surface expression of the adenosine 2b receptor in intestinal epithelial cells: role of SNARE proteins, Am. J. Physiol. Gastrointest. Liver Physiol., 287, G1100–G1107.
Rajagopal, M., and Pao, A. C. (2010) Adenosine activates a2b receptors and enhances chloride secretion in kidney inner medullary collecting duct cells, Hypertension, 55, 1123–1128.
Berglindh, T., and Obrink, K. J. (1976) A method for preparing isolated glands from the rabbit gastric mucosa, Acta Physiol. Scand., 96, 150–159.
Fryklund, J., Wallmark, B., Larsson, H., and Helander, H. F. (1984) Effect of omeprazole on gastric secretion in H+,K+-ATPase and in pepsinogen-rich cell fractions from rabbit gastric mucosa, Biochem. Pharmacol., 33, 273–280.
Arin, R. M., Rueda, Y., Casis, O., Gallego, M., Vallejo, A. I., and Ochoa, B. (2014) Basolateral expression of GRP94 in parietal cells of gastric mucosa, Biochemistry (Moscow), 79, 8–15.
Muallem, S., Burnham, C., Blissard, D., Berglindh, T., and Sachs, G. (1985) Electrolyte transport across the basolateral membrane of the parietal cells, J. Biol. Chem., 260, 6641–6653.
Aran, J. M., Colomer, D., Matutes, E., Vives-Corrons, J. L., and Franco, R. (1991) Presence of adenosine deaminase on the surface of mononuclear blood cells: immunochemical localization using light and electron microscopy, J. Histochem. Cytochem., 39, 1001–1008.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 72, 248–254.
Heldsinger, A. A., Vinik, A. I., and Fox, I. H. (1986) Inhibition of guinea-pig oxyntic cell function by adenosine and prostaglandins, J. Pharmacol. Exp. Ther., 237, 351–356.
Gerber, J. G., Nies, A. S., and Payne, N. A. (1985) Adenosine receptors on canine parietal cells modulate gastric acid secretion to histamine, J. Pharmacol. Exp. Ther., 233, 623–627.
Yip, L., and Kwok, Y. N. (2004) Role of adenosine A2A receptor in the regulation of gastric somatostatin release, J. Pharmacol. Exp. Ther., 309, 804–815.
Yang, G. K., Chen, J. F., Kieffer, T. J., and Kwok, Y. N. (2009) Regulation of somatostatin release by adenosine in the mouse stomach, J. Pharmacol. Exp. Ther., 329, 729–737.
Tan-Allen, K. Y., Sun, X. C., and Bonanno, J. A. (2005) Characterization of adenosine receptors in bovine corneal endothelium, Exp. Eye Res., 80, 687–696.
Olanrewaju, H. A., Qin, W., Feoktistov, I., Scemama, J. L., and Mustafa, S. J. (2000) Adenosine A(2A) and A(2B) receptors in cultured human and porcine coronary artery endothelial cells, Am. J. Physiol. Heart Circ. Physiol., 279, H650–656.
Linden, J., Thai, T., Figler, H., Jin, X., and Robeva, A. S. (1999) Characterization of human A(2B) adenosine receptors: radioligand binding, Western blotting, and coupling to G(q) in human embryonic kidney 293 cells and HMC-1 mast cells, Mol. Pharmacol., 56, 705–713.
Richard, E., Alam, S. M., Arredondo-Vega, F. X., Patel, D. D., and Hershfield, M. S. (2002) Clustered charged amino acids of human adenosine deaminase comprise a functional epitope for binding the adenosine deaminase complexing protein CD26/dipeptidyl peptidase IV, J. Biol. Chem., 277, 19720–19726.
Beraudi, A., Traversa, U., Villani, L., Sekino, Y., Nagy, J. I., and Poli, A. (2003) Distribution and expression of A1 adenosine receptors, adenosine deaminase and adenosine deaminase-binding protein (CD26) in goldfish brain, Neurochem. Int., 42, 455–464.
Sakai, H., Okada, Y., Morii, M., and Takeguchi, N. (1989) Anion and cation channels in the basolateral membrane of rabbit parietal cells, Pflugers Arch., 414, 185–192.
Karam, S. M. (2010) A focus on parietal cells as a renewing cell population, World J. Gastroenterol., 16, 538–546.
Chew, C. S. (1994) Parietal cell culture: new models and directions, Annu. Rev. Physiol., 56, 445–461.
Miller, M. L., Andringa, A., Zavros, Y., Bradford, E. M., and Shull, G. E. (2010) Volume density, distribution, and ultrastructure of secretory and basolateral membranes and mitochondria predict parietal cell secretory (dys)function, J. Biomed. Biotechnol., 2010, 394198.
Chew, C. S., Chen, X., Parente, J. A., Jr., Tarrer, S., Okamoto, C., and Qin, H. Y. (2002) Lasp-1 binds to nonmuscle F-actin in vitro and is localized within multiple sites of dynamic actin assembly in vivo, J. Cell. Sci., 115, 4787–4799.
Lapierre, L. A., Avant, K. M., Caldwell, C. M., Ham, A. J., Hill, S., Williams, J. A., Smolka, A. J., and Goldenring, J. R. (2007) Characterization of immunoisolated human gastric parietal cells tubulovesicles: identification of regulators of apical recycling, Am. J. Physiol. Gastrointest. Liver Physiol., 292, G1249–1262.
Geibel, J., Abraham, R., Modlin, I., and Sachs, G. (1995) Gastrin-stimulated changes in Ca2+ concentration in parietal cells depends on adenosine 3′,5′-cyclic monophosphate levels, Gastroenterology, 109, 1060–1067.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published in Russian in Biokhimiya, 2015, Vol. 80, No. 1, pp. 150–156.
Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM14-219, December 28, 2014.
Rights and permissions
About this article
Cite this article
Arin, R.M., Vallejo, A.I., Rueda, Y. et al. The A2B adenosine receptor colocalizes with adenosine deaminase in resting parietal cells from gastric mucosa. Biochemistry Moscow 80, 120–125 (2015). https://doi.org/10.1134/S0006297915010149
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0006297915010149