Adenosine A2a receptor-mediated inhibition of rod opsin mRNA expression in tiger salamander

The neuromodulator adenosine mediates dark-adaptive changes in retinal photoreceptors through A(2a) receptors. In cold-blooded vertebrates, opsin mRNA expression is lower at night than during the day. In the present study, we tested whether adenosine could inhibit opsin mRNA expression in cultured rod cells and if endogenous adenosine acts to suppress opsin mRNA in the intact retina at night. Semi-quantitative in situ hybridization showed that treatment with 100 nm of the A(2a) /A(2b) agonist N (6) -[2-(3,5-dimethoxyphenyl)-2-(2-methylphenyl)-ethyl]adenosine (DPMA) reduced opsin mRNA 41% in cultured rod cells. The effect of DPMA was blocked by 10 mum of the A(2a) antagonist 8-(3-chlorostyryl)caffeine (CSC) but not by 10 mum of the A(2b) antagonist alloxazine. One micromolar adenosine alone had no effect on opsin mRNA. However, in the presence of the adenosine deaminase inhibitor erythro -9-(2-hydroxy-3-nonyl)adenine hydrochloride (EHNA), 1 mum adenosine reduced opsin mRNA 61%. EHNA alone reduced opsin mRNA by 26%. Consistent with an A(2a) receptor mechanism, 100 nm forskolin (adenylate cyclase agonist) decreased opsin mRNA 34%. Finally, northern blots showed that intravitreal injection of 10 mum CSC at night increased opsin I mRNA 38%. Thus, endogenous adenosine suppresses rod opsin I mRNA expression at night; in vitro results indicate this reduction occurs through A(2a) -like receptor binding and stimulation of adenylate cyclase activity.