When tested under conditions reducing the endogenous production of adenosine (presence of adenosine deaminase (ADA) 1.6 IU/ml; and deoxyadenosine triphosphate (d-ATP), and in the presence of both NaCl and GTP, the ADA-resistant analog phenylisopropyladenosine (PIA) inhibited the adenylate cyclase of several brain tissues. These tissues included: (1) 5 brain areas of adult rats (frontal and parietal cortex, cerebellum cortex, hippocampus and striatum)--hypothalamus and mid-brain adenylate cyclases were not inhibited by PIA; (2) astrocytes in primary cultures prepared from cerebral cortex of newborn mice; and (3) neurons in primary cultures prepared from striata of 15-day-old mouse embryos. The specificity profile of the adenosine receptor involved in the inhibition was determined in astrocytes. It was typical of an A1 adenosine receptor (high affinity of PIA; Ka app: 9 +/- 5 X 10(-9) M (n = 4) compared to the affinity of 5'-N-ethylcarboxamide adenosine (NECA); Ka app: 1.3 +/- 0.6 X 10(-7) M (n = 3). There was an excellent correlation between the affinities of several adenosine agonists and antagonists for A1 receptors coupled with an adenylate cyclase in astrocytes and for the receptors labeled with N6-cyclohexyl-[3H]adenosine in brain cortex. In adult rat striatum as well as in astrocytes and striatal neurons in culture the adenylate cyclase was inhibited by low PIA concentrations through A1 receptors and stimulated by higher concentrations through A2 receptors. In contrast, A2 receptors were not detected in adult rat cerebral cortex. In adult rat striatum, A1 and dopamine receptors coupled with an adenylate cyclase seemed to be located on different cell populations. In contrast, in astrocytes A1 and beta-adrenergic receptors coupled with adenylate cyclase were apparently located on the same cells.