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Adenosine A1 Receptor-Dependent Antinociception Induced by Inosine in Mice: Pharmacological, Genetic and Biochemical Aspects

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Abstract

Inosine is an endogenous nucleoside that has anti-inflammatory and antinociceptive properties. Inosine is a metabolite of adenosine, and some of its actions suggest the involvement of adenosine A1 receptors (A1Rs). The purpose of this study was to better understand mechanisms of inosine-induced antinociception by investigating the role of A1Rs and purine metabolism inhibitors. Inosine antinociception was evaluated using the formalin test in mice. An A1R-selective antagonist (DPCPX), A1R knockout mice (gene deletion) and mice with A1R reduced expression (antisense oligonucleotides) were used to assess the role of A1Rs in the antinociceptive action of inosine. Binding assays were performed to compare the affinity of inosine and adenosine for A1Rs. Finally, the role of adenosine and inosine breakdown was assessed using deoxycoformycin (DCF) and forodesine (FDS) as enzymatic inhibitors of adenosine deaminase and purine nucleoside phosphorylase, respectively. Inosine induced antinociception in the formalin test when given by systemic, spinal and peripheral routes. Systemically, inosine exhibited a potency similar to adenosine, and its effects were inhibited by DPCPX. Inosine did not induce antinociception in A1R knockout mice or in mice with reduced A1R expression. In binding studies, inosine bound to A1Rs with an affinity similar to adenosine. DCF had no effect on inosine actions. FDS augmented the antinociceptive effect of a low systemic dose of inosine and, at a higher dose, induced antinociception by itself. Collectively, these data indicate that inosine is an agonist for A1Rs with antinociceptive properties and a potency similar to adenosine and can be considered another endogenous ligand for this receptor.

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Acknowledgments

This study was supported by grants from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina (FAPESC), Brazil. We wish to thank Bertil Fredholm (Karolinska Institute, Stockholm, Sweden) who supplied the initial adenosine A1R knockout mice (to JS), and whose laboratory performed polymerase chain reaction genotyping. Also, we would like to thank BioCryst Pharmaceuticals Inc. and Dr. ShantaBantia for the donation of forodesine. F.P.N., S.J.M.J., F.A.P. and A.R.S.S. thank the CNPq for their fellowship support.

Conflict of Interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Laboratório de Neurobiologia da Dor e Inflamação, Departamento de Ciências Fisiológicas, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil

    Francisney Pinto Nascimento, Sérgio José Macedo-Júnior, Murilo Luiz-Cerutti, Marina Machado Córdova & Adair Roberto Soares Santos

  2. Programa de Pós-Graduação em Farmacologia, Departamento de Farmacologia, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil

    Francisney Pinto Nascimento, Sérgio José Macedo-Júnior, Rafael Cypriano Dutra & Adair Roberto Soares Santos

  3. Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Santa Catarina, Campus Universitário, Trindade, Florianópolis, SC, 88040-900, Brazil

    Fabrício Alano Pamplona, Leandra Constantino & Carla Inês Tasca

  4. Instituto D’Or de Pesquisa e Ensino (IDOR), Rio de Janeiro, RJ, 22230-100, Brazil

    Fabrício Alano Pamplona

  5. Centro de Inovação e Ensaios Pré-clínicos, Cachoeira do Bom Jesus, Florianópolis, SC, 88056-000, Brazil

    João B. Calixto

  6. Department of Pharmacology, Dalhousie University, Halifax, NS, Canada

    Allison Reid & Jana Sawynok

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  1. Francisney Pinto Nascimento
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  2. Sérgio José Macedo-Júnior
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  5. Marina Machado Córdova
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  10. Allison Reid
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  12. Adair Roberto Soares Santos
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Correspondence to Adair Roberto Soares Santos.

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Nascimento, F.P., Macedo-Júnior, S.J., Pamplona, F.A. et al. Adenosine A1 Receptor-Dependent Antinociception Induced by Inosine in Mice: Pharmacological, Genetic and Biochemical Aspects. Mol Neurobiol 51, 1368–1378 (2015). https://doi.org/10.1007/s12035-014-8815-5

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