OBJECTIVE: The aim of this study is to investigate the role of the purinergic P2Y2 receptor in learning and memory processes. MATERIALS AND METHODS: Behavioral, electrophysiological, and biochemical tests of memory function were conducted in P2Y2 receptor knockout (P2Y2R-KO) mice, and the findings were compared to those of wild-type mice with the help of unpaired Student’s t-test. RESULTS: The findings of the behavioral Y-maze test showed that the P2Y2R-KO mice had impaired memory and cognitive function. Electrophysiological studies on paired-pulse facilitation showed that glutamate release was higher in the P2Y2R-KO mice than in the WT mice. Furthermore, PCR and Western blot analysis revealed that the mRNA and protein expression of acetylcholinesterase E (AChE) and alpha-7 nicotinic acetylcholine receptors (α7 nAChRs) were increased in the hippocampus of P2Y2R-KO mice. CONCLUSIONS: The findings of this study indicate that P2Y2 receptors are important regulators of both glutamatergic and cholinergic systems in the hippocampus. © 2020 Verduci Editore s.r.l. All rights reserved.