Abstract
1. The potential neuroprotective actions of the A3 adenosine receptor (A3AR) were investigated using mice with functional deletions of the A3AR (A3AR−/−) in behavioral assessments of analgesia, locomotion, tests predictive of depression and anxiety, and the effects of mild hypoxia on cognition and neuronal survival.
2. Untreated A3AR−/− mice were tested in standard behavioral paradigms, including activity in the open field, performance in the hot-plate, tail-flick, tail-suspension, and swim tests, and in the elevated plus maze. In addition, mice were exposed repeatedly to a hypoxic environment containing carbon monoxide (CO). The cognitive effects of this treatment were assessed using the contextual fear conditioning test. After testing, the density of pyramidal neurons in the CA1, 2, and 3 subfields of the hippocampus was determined using standard histological and morphometric techniques.
3. A3AR−/− mice showed increased locomotion in the open field test, elevated plus maze (number of arm entries) and light/dark box (number of transitions). However, they spent more time immobile in two different tests of antidepressant activity (Swim and tail suspension tests). A3AR−/− mice also showed evidence of decreased nociception in the hot-plate, but not tail-flick tests. Further, A3AR−/− mice were more vulnerable to hippocampal pyramidal neuron damage following episodes of carbon monoxide (CO)-induced hypoxia. One week after exposure to CO a moderate loss of pyramidal neurons was observed in all hippocampal subfields of both wild-type (A3AR+/+) and A3AR−/− mice. However, the extent of neuronal death in the CA2–3 subfields was less pronounced in A3AR+/+ than A3AR−/− mice. This neuronal loss was accompanied by a decline in cognitive function as determined using contextual fear conditioning. These histological and cognitive changes were reproduced in wild-type mice by repeatedly administering the A3AR-selective antagonist MRS 1523 (5-propyl-2-ethyl-4-propyl-3-(ethylsulfanylcarbonyl)-6-phenylpyridine-5-carboxylate 1 mg/kg i.p.).
4. These results indicate that pharmacologic or genetic suppression of A3AR function enhances some aspects of motor function and suppresses pain processing at supraspinal levels, while acting as a depressant in tests predictive of antidepressant action. Consistent with previous reports of the neuroprotective actions of A3AR agonists, A3AR−/− mice show an increase in neurodegeneration in response to repeated episodes of hypoxia.
Similar content being viewed by others
References
Appel, E., Kazimirsky, G., Ashkenazi, E., Kim, S. G., Jacobson, K. A., and Brodie, C. (2001). Roles of BCL-2 and caspase 3 in the adenosine A3 receptor-induced apoptosis. J. Mol. Neurosci. 17:285–292.
Brand, A., Vissiennon, Z., Eschke, D., and Nieber, K. (2001). Adenosine A(1) and A(3) receptors mediate inhibition of synaptic transmission in rat cortical neurons. Neuropharmacology 40:85–95.
Cerniway, R. J., Yang, Z., Jacobson, M. A., Linden, J., and Matherne, G. P. (2001). Targeted deletion of A3 adenosine receptors improves tolerance to ischemia-reperfusion injury in mouse myocardium. Am. J. Physiol. 281:H1751-H1758.
Di Iorio, P., Kleywegt, S., Ciccarelli, R., Traversa, U., Andrew, C. M., Crocker, C. E., Werstiuk, E. S., and Rathbone, M. P. (2002). Mechanisms of apoptosis induced by purine nucleosides in astrocytes. Glia 38:179.
Dunwiddie, T. V., Diao, L., Kim, H. O., Jiang, J.-l., and Jacobson, K. A. (1997). Activation of hippocampal adenosine A3 receptors produces a heterologous desensitization of A1 receptor mediated responses in rat hippocampus. J. Neurosci. 17:607–614.
El Yacoubi, M., Ledent, C., Parmentier, M., Bertorelli, R., Ongini, E., Costentin, J., and Vaugeois, J. M. (2001). Adenosine A2A receptor antagonists are potential antidepressants: Evidence based on pharmacology and A2A receptor knockout mice. Br. J. Pharmacol. 134:68–77.
Feoktistov, I., Goldstein, A. E., Ryzhov, S., Zeng, D., Belardinelli, L., Voyno-Yasenetskaya, T., and Biaggioni, I. (2002). Differential expression of adenosine receptors in human endothelial cells: Role of A2B receptors in angiogenic factor regulation. Circ. Res. 90:531–538.
Fishman, P., Madi, L., Bar-Yehuda, S., Barer, F., Del Valle, L., and Khalili, K. (2002). Evidence for involvement of Wnt signaling pathway in IB-MECA mediated suppression of melanoma cells. Oncogene 21:4060–4064.
Fredholm, B. B., IJzerman, A. P., Jacobson, K. A., Klotz, K. N., and Linden, J. (2001). International Union of Pharmacology. XXV. Nomenclature and classification of adeonsine receptors. Pharmacol. Rev. 53:527–552.
Gimenez-Llort, L., Fernandez-Teruel, A., Escorihuela, R. M., Fredholm, B. B., Tobena, A., Pekny, M., and Johansson, B. (2002). Mice lacking the adenosine A1 receptor are anxious and aggressive, but are normal learners with reduced muscle strength and survival rate. Eur. J. Neurosci. 16:547–550.
Guo, Y., Bolli, R., Bao, W., Wu, W.-J., Black, R. G. Jr., Murphree, S. S., Salvatore, C. A., Jacobson, M. A., and Auchampach, J. A. (2001). Targeted deletion of the adenosine A3 receptor confers resistance to myocardial ischemic injury and does not prevent early preconditioning. J. Mol. Cell Cardiol. 33:825–830.
Harrison, G. J., Cerniway, R. J., Peart, J., Berr, S. S., Ashton, K., Regan, S., Paul, Matherne, G., and Headrick, J. P. (2002). Effects of A(3) adenosine receptor activation and gene knock-out in ischemic-reperfused mouse heart. Cardiovasc Res. 53:147.
Ishimaru, H., Nabeshima, T., Katon, A., Suzuki, H., Fukuta, T., and Kameyama, T. (1991). Effect of successive carbon monoxide exposures on delayed neuronal death in mice under the maintenance of normal body temperature. Biochem. Biophys. Res. Commun 179:836–840.
Jacobson, K. A. (1998). Adenosine A3 receptors: Novel ligands and paradoxical effects. Trends Pharmacol. Sci. 19:184–191.
Jacobson, K. A., Nikodijevic, O., Shi, D., Gallo-Rodriguez, C., Olah, M. E., Stiles, G. L., and Daly, J. W. (1993). A role for central A3-adenosine receptors: Mediation of behavioral depressant effects. FEBS Lett. 336:57–60.
Jacobson, K. A., Park, K.-S., Jiang, J.-L., Kim, Y.-C., Olah, M. E., Stiles, G. L., and Ji, X.-D. (1997). Pharmacological characterization of novel A3 adenosine receptor-selective antagonists. Neuropharmacology 36:1157–1165.
Kustova, Y., Sung, E.-G., Morse, D., Sei, Y., and Basile, A. (1999). Histological evidence of neuronal degeneration in mice infected with LP-BM5 murine leukemia virus. Mol. Chem. Neuropathol. 35:39–59.
Lasley, R. D., Narayan, P., Jahania, M. S., Partin, E. L., Kraft, K. R., and Mentzer, R. M. Jr. (1999). Species-dependent hemodynamic effects of adenosine A3-receptor agonists IB-MECA and Cl-IB-MECA. Am. J. Physiol. 276:H2076-H2084.
Macek, T. A., Schaffhauser, H., and Conn, P. J. (1998). Protein kinase C and A3 adenosine receptor activation inhibit presynaptic metabotropic glutamate receptor (mGluR) function and uncouple mGluRs from GTP-binding proteins. J. Neurosci. 18:6138–6146.
Maurice, T., Hiramatsu, M., Kameyama, T., Hasegawa, T., and Nabeshima, T. (1994). Behavioral evidence for a modulating role of sigma ligands in memory processes. II. Reversion of carbon monoxide-induced amnesia. Brain Res. 647:57–64.
Maurice, T., Phan, V. L., Noda, Y., Yamada, K., Privat, A., Nabeshima, T. (1999). The attenuation of learning impairments induced after exposure to CO or trimethyltin in mice by sigma (sigma) receptor ligands involves both sigma1 and sigma2 sites. Br. J. Pharmacol. 127:335–342.
Nabeshima, T., Katon, A., Ishimaru, H., Yoneda, Y., Ogita, K., Murase, K., Ohtsuka, H., Inari, K., Fukuta, T., and Kameyama, T. (1991). Carbon monoxide-induced delayed amnesia, delayed neuronal death and change in acetylcholine concentration in mice. J. Pharmacol. Exp. Ther. 256: 378–384.
Okada, M., Kawata, Y., Murakami, T., Wada, K., Mizuno, K., Kondo, T., and Kaneko, S. (1999). Differential effects of adenosine receptor subtypes on release and reuptake of hippocampal serotonin. Eur. J. Neurosci. 11:1–9
Parsons, M., Young, L., Lee, J.-E., Jacobson, K. A., and Liang, B. T. (2000). Distinct cardioprotective effects of adenosine mediated by differential coupling of receptor subtypes to phospholipases C and D. FASEB J. 14:1423–1431.
Porsolt, R. D., Bertin, A., Blavet, N., Deniel, M., and Jalfre, M. (1979). Immobility induced by forced swimming in rats: Effects of agents which modify central catecholamine and serotonin activity. Eur. J. Pharmacol. 57:201–210.
Ramkumar, V., Stiles, G. L., Beaven, M. A., and Ali, H. (1993). The A3 adenosine receptor is the unique adenosine receptor which facilitates release of allergic mediators in mast cells. J. Biol. Chem. 268:16887–16890.
Rivkees, S. A., Thevananther, S., and Hao, H. (2000). Are A3 adenosine receptors expressed in the brain? Neuroreport 11:1025–1030.
Sajjadi, F. G., Takabayashi, K., Foster, A. C., Domingo, R. C., and Firestein, G. S. (1996). Inhibition of TNF-alpha expression by adenosine: role of A3 adenosine receptors. J. Immunol. 156:3435–3442.
Salvatore, C. A., Tilley, S. L., Latour, A. M., Fletcher, D. S., Koller, B. H., and Jacobson, M. A. (2000). Disruption of the A3 adenosine receptor gene in mice and its effect on stimulated inflammatory cells. J. Biol. Chem. 275:4429–4434.
Shepherd, R. K., Linden, J., and Duling, B. R. (1996). Adenosine-induced vasoconstriction in vivo. Role of the mast cell and A3 adenosine receptor. Circ. Res. 78:627–634.
Sterneck, E., Paylor, R., Jackson-Lewis, V., Libbey, M., Przedborski, S., Tessarollo, L., Crawley, J., and Johnson, P. (1998). Selectively enhanced contextual fear conditioning in mice lacking the transcriptional regulator CCAAT/enhancer binding protein sigma. Proc. Natl. Acad. Sci. U.S.A. 95:10908–10913.
Steru, L., Chermat, R., Thierry, B., and Simon, P. (1985). The tail suspension test: A new method for screening antidepressants in mice. Psychopharmacology 85:367–370.
Talukder, M. A., Morrison, R. R., Jacobson, M. A., Jacobson, K. A., Ledent, C., and Mustafa, S. J. (2002). Targeted deletion of adenosine A(3) receptors augments adenosine-induced coronary flow in isolated mouse heart. Am. J. Physiol. Heart. Circ. Physiol. 282:H2183-H2189.
van Schaick, E. A., Jacobson, K. A., Kim, H. O., IJzerman, A. P., and Danhof, M. (1996). Hemodynamic effects and histamine release elicited by the selective adenosine A3 receptor agonist 2-Cl-IB-MECA in conscious rats. Eur. J. Pharmacol. 308:311–314.
von Lubitz, D. K. J. E., Lin, R. C.-S., Popik, P., Carter, M. F., and Jacobson, K. A. (1994). Adenosine A3 receptor stimulation and cerebral ischemia. Eur. J. Pharmacol. 263:59–67.
Yaar, R., Lamperti, E. D., Toselli, P. A. Ravid, K. (2002). Activity of the A3 adenosine receptor gene promoter in transgenic mice: Characterization of previously unidentified sites of expression. FEBS Lett. 532:267–272.
Zhao, Z., Makaritsis, K., Francis, C. E., Gavras, H., and Ravid, K. (2000). A role for A3 adenosine receptor in determining tissue levels of cAMP and blood pressure: Studies in knock-out mice. Biochem. Biophys. Acta 1500:280–290.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Fedorova, I.M., Jacobson, M.A., Basile, A. et al. Behavioral Characterization of Mice Lacking the A3 Adenosine Receptor: Sensitivity to Hypoxic Neurodegeneration. Cell Mol Neurobiol 23, 431–447 (2003). https://doi.org/10.1023/A:1023601007518
Issue Date:
DOI: https://doi.org/10.1023/A:1023601007518