Recent　studies　indicate　that　presynaptic　long-term　potentiation　in　the　anterior　cingulate　cortex　may　contribute　to　chronic　pain-related　anxiety.　In　addition　to　the　anterior　cingulate　cortex,　the　insular　cortex　has　also　been　indicated　in　chronic　pain　and　its　related　emotional　disorders.　In　the　present　study,　we　used　a　64-channel　multielectrode　dish　(MED64)　system　to　record　pre-long-term　potentiation　in　the　insular　cortex.　We　showed　that　low-frequency　stimulation　paired　with　a　GluK1-containing　kainate　receptor　agonist　induced　N-methyl-D-aspartic　acid　receptor-independent　pre-long-term　potentiation　in　the　insular　cortex　of　wild-type　mice.　This　form　of　pre-long-term　potentiation　was　blocked　in　the　insular　cortex　of　adenylyl　cyclase　subtype　1　(AC1)　knockout　mice.　Furthermore,　a　selective　AC1　inhibitor　NB001　blocked　pre-long-term　potentiation　in　the　insular　cortex　with　a　dose-dependent　manner.　Taken　together,　our　results　suggest　that　AC1　contributes　to　pre-long-term　potentiation　in　the　insular　cortex　of　adult　mice　and　NB001　may　produce　anxiolytic　effects　by　inhibiting　pre-long-term　potentiation　in　the　anterior　cingulate　cortex　and　insular　cortex.