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Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications
Published on: January 27, 2014
Different modulation of inhibitory and stimulatory pathways mediated by adenosine after chronic in vivo agonist
María Angeles Ruiz1, José Luis Albasanz, David León
1Area de Bioquímica, Departamento de Química Inorgánica, Orgánica y Bioquímica, Facultad de Químicas, Centro Regional de Investigaciones Biomédicas, Universidad de Castilla-La Mancha, Avenida Camilo José Cela, 10. 13071 Ciudad Real, Spain.
Abstract:
After 6 days of in vivo treatment with two selective adenosine receptor agonists, 5'-N-ethylcarboxamido adenosine (NECA) and R-N6-phenylisopropiladenosine (R-PIA), we investigated their effects on adenosine receptors/adenylyl cyclase system in synaptic plasma membranes isolated from rat brain. NECA treatment caused a significant loss of NECA-stimulated adenylyl cyclase activity, suggesting a desensitization of the adenosine A2 receptors-mediated pathway. No significant differences in total adenosine A2 receptors were observed, but Gs protein levels were decreased, suggesting Gs down-regulation as a mechanism for desensitization. On the other hand, NECA treatment caused a significant decrease in high-affinity adenosine A1 receptors population; however, no changes in CHA-inhibited adenylyl cyclase activity or Gi protein level were observed. Finally, when we studied the effects of R-PIA, a selective adenosine A1 receptor agonist, on stimulatory pathway of adenosine, low-affinity adenosine A2 binding sites were decreased without affecting the functionality of the pathway. These results show that adenosine A1 and A2 receptors are modulated in a different way after chronic agonist exposure and suggest the existence of cross-talk mechanisms between both stimulatory an inhibitory pathways mediated by adenosine.
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