Adenosine, which accumulates rapidly during ischemia due to the breakdown of ATP, has beneficial effects in many tissues. We examined whether adenosine induces the production of glial cell line-derived neurotrophic factor (GDNF) in cultured astrocytes. We evaluated GDNF mRNA expression and GDNF production in astrocytes cultured with adenosine and the adenosine selective receptor agonists 5-( N-ethylcarboxamido) adenosine (NECA), N 6-cyclopentyladenosine (CPA) and 2- p-(2-carboxyethyl) phenethylamino-5′- N-ethylcarboxamindo-adenosine hydrochloride (CGS 21680). Moreover, we examined the possibility that the expression of GDNF is regulated differently in cultured astrocytes from the stroke-prone spontaneously hypertensive rat (SHRSP) than in those from Wistar Kyoto rats (WKY). In this study, we confirmed that adenosine and the selective A 2B adenosine receptor agonist NECA induced the expression of GDNF in cultured astrocytes. The A 2B receptor antagonist alloxazine was able to inhibit the increase in extracellular GDNF produced by adenosine. Furthermore, the amounts of GDNF produced were significantly reduced in astrocytes of the adenosine-treated SHRSP compared with those of WKY. These results indicate that adenosine induces the expression of GDNF, and adenosine A 2B receptors participate in the regulation of GDNF levels in astrocytes. This expression was attenuated in astrocytes of SHRSP compared with those of WKY.