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Stimulation of α2A-adrenoceptors (ARs) in the prefrontal cortex (PFC) produces a beneficial effect on cognitive functions such as working memory.The α2A-adrenergic agonist guanfacine has been used for treatment of psychiatric disorders such as attention-deficit/hyperactivity disorder (ADHD).A previous study in our laboratory showed that stimulation of α2A-AR suppresses excitatory synaptic transmission in pyramidal cells of the rat medial prefrontal cortex (mPFC).However, the intracellular mechanism underlying the α2A-AR suppression remains unclear.In the present study, we recorded evoked excitatory postsynaptic current (eEPSC) in layer Ⅴ-Ⅵ pyramidal cells of the mPFC, using whole-cell patch-clamp recording technique, and found that the α2A-AR inhibition of excitatory synaptic transmission is mediated via Gi-cAMP-PKA-PP 1 and/or Gi-cAMP-PKA-PP 1-CaMKII signaling pathways.