OBJECTIVE Phosphodiesterase 4(PDE4),specific for cyclicAMP(cAMP)-hydrolyzing,has four isoforms(PDE4A-D) with at least 25 splice variants. PDE4 inhibitors produce definite antidepressant-like and cognitive-enhancing effects. However,none of PDE4 inhibitors has yet been approved for clinical utility so far due to the concomitant side effects. The present research is to explore the splice variants of PDE4 D responsible for antidepressant-like and cognitive-enhancing effects of PDE4 inhibitors but not side effects. METHODS Long-form PDE4 Ds were silenced by the bilateral microinfusion of lentiviral vector containing mi RNAs(4Dmi R) into the prefrontal cortex(PFC),PDE4D4 or D5 was overexpressed by the bilateral microinfusion of lentiviral vector containing full c DNA into hippocampus. Antidepressant-like behaviors were measured by tail-suspension test(TST),forced swimming test(FST)and chronic unpredictable stress model. Cognitive behaviors were measured by the novel object recognition test(NOR) and Morris water maze test(MWM) in both normal mice and the mice with chronic unpredictable stress-induced memory deficits. The emetic potential was evaluated by the assessment of the anaesthetic reversal effect,a surrogate of the emesis test in non-vomiting species. The expressions of PDE4 isoforms/splice variants and cAMP level were examined by Western-blot and ELISA analysis. The dendritic complexity and spine density were assessed by Golgi staining. RESULTS(1)High and specific expression of EGFP(green,indicator of 4Dmi R expression) in PFC was observed under fluorescence microscopy.(2) 4Dmi R significantly down-regulated PDE4D4/5 splice variants,but not PDE4 A,PDE4 B or PDE4D1/2/3.(3) 4Dmi R treatments significantly increased cAMP signaling and dendritic complexity in PFC.(4) Rolipram and/or 4Dmi R treatments significantly decreased immobility in TST and FST.(5) Rolipram and/or 4Dmi R treatments reversed the depressive-like behaviors in chronically stressed mice,including the reduced sucrose preference,prolonged latency to novelty-suppressed feeding and increased immobility in FST.(6) Rolipram and/or 4Dmi R treatments significantly increased the recognition index in NOR task and both the entries and durations in MWM task.(7) Rolipram and/or 4Dmi R treatments reversed the memory deficits in chronically stressed mice,including the reduced the recognition index in NOR task and the decreased durations in MWM task.(8) Rolipram and/or 4DmiR treatments reversed the decreased cA MP signaling,dendritic complexity and spine density.(9) Rolipram or plus 4Dmi R treatment significantly decreased the duration of anaesthesia in the alpha2 adrenergic receptor-mediated anesthesia,but not 4Dmi R treatment alone.(10)Hippocampal overexpression of PDE4D5,but not PDE4D4,produced depressive-like and cognitive defect behaviors,which were reversed by rolipram.The measurements including cAMP signaling,dendritic complexity and in vivo hippocampal LTP,showed the same changes. CONCLUSION Long-form PDE4 Ds,especially the PDE4D5,are the major isoforms responsible for antidepressant-like and cognitive-enhancing effects with little side effects. The critical roles of long-form PDE4 Ds are mediated by their regulation of cAMP signaling pathway and neuroplasticity. OBJECTIVE Phosphodiesterase 4 (PDE4), specific for cyclic AMP (cAMP) -hydrolyzing, has four isoforms (PDE4A-D) with at least 25 splice variants. PDE4 inhibitors produce definite antidepressant-like and cognitive- enhancing effects. However, none of PDE4 inhibitors has yet been approved for clinical utility so far due to the concomitant side effects. The present research is to explore the splice variants of PDE4 D responsible for antidepressant-like and cognitive-enhancing effects of PDE4 inhibitors but not side effects. PDE4 Ds were silenced by the bilateral microinfusion of lentiviral vector containing mi RNAs (4Dmi R) into the prefrontal cortex (PFC), PDE4D4 or D5 was overexpressed by the bilateral microinfusion of lentiviral vector containing full c DNA into hippocampus. Antidepressant-like behaviors were measured by tail-suspension test (TST), forced swimming test (FST) and chronic unpredictable stress model. Cognitive behaviors were measured by the novel object recognition test ( NOR) and Morris water maze test (MWM) in both normal mice and the mice with chronic unpredictable stress-induced memory deficits. The emetic potential was evaluated by the assessment of the anaesthetic reversal effect, a surrogate of the emesis test in non-vomiting The expressions of PDE4 isoforms / splice variants and cAMP level were examined by Western-blot and ELISA analysis. The dendritic complexity and spine density were assessed by Golgi staining. RESULTS (1) High and specific expression of EGFP (green, indicator of (2) 4Dmi R significantly down-regulated PDE4D4 / 5 splice variants, but not PDE4 A, PDE4 B or PDE4D1 / 2 / 3. (3) 4Dmi Rsories significantly increased cAMP signaling and dendritic complexity in PFC. (4) Rolipram and / or 4Dmi R treatments significantly decreased immobility in TST and FST. (5) Rolipram and / or 4Dmi Rsense the depressive-like behaviors in chronically stressed mice, including thereduced sucrose preference, prolonged latency to novelty-suppressed feeding and increased immobility in FST. (6) Rolipram and / or 4Dmi R treatments were significantly increased the recognition index in NOR task and both the entries and durations in MWM task. (7) Rolipram and / or 4Dmi R radicals that have reduced the recognition index in NOR task and the decreased durations in MWM task. (8) Rolipram and / or 4DmiR induced reversed the decreased cA MP signaling, dendritic complexity and (9) Rolipram or plus 4Dmi R treatment significantly decreased the duration of anaesthesia in the alpha2 adrenergic receptor-mediated anesthesia, but not 4Dmi R treatment alone. (10) Hippocampal overexpression of PDE4D5, but not PDE4D4, produced depressive-like and cognitive defect behaviors, which had reversed ciproli. The measurements include cAMP signaling, dendritic complexity and in vivo hippocampal LTP, showed the same changes. CONCLUSI ON long-form PDE4 Ds, especially the PDE4D5, are the major isoforms responsible for antidepressant-like and cognitive-enhancing effects with little side effects. The critical roles of long-form PDE4 Ds are mediated by their regulation of cAMP signaling pathway and neuroplasticity .