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The illuminated compartment contained a 50 W bulb, and its oor was composed of 2 mm stainless steel rods spaced with centres 1 cm apart. A mouse was initially placed inside the illuminated compartment to the acquisition trial, and also the door between the two compartments was opened 10 s later.

Memory impairment was induced by diazepam, a selective antagonist in the benzodiazepine internet site in the GABAA receptor or MK 801, an NMDA receptor channel blocker, which was administered 10 min following tanshinone I or vehicle. Manage animals had been administered vehicle option only. Twenty four hours following a single acquisition trial, the histone deacetylase inhibitor mice were subjected to retention trial and placed again in the illuminated compartment. The times taken for a mouse to enter the dark compartment after door IEM 1754 opening was dened as latency time for both acquisition and retention trials. Latency to enter the dark compartment was recorded for up to 300 s. To investigate the eect of tanshinone I alone on memory, tanshinone I was given to mice 40 min before the acquisition trial. To avoid a ceiling eect in unimpaired animals, foot shock intensity was set at 0.

c. v. injection and anaesthetic IEM 1754 agents also aects those parameters. In the present study, we measured the spontaneous locomotor behaviour, as described previously, to assess whether the anaesthetic agent or stress by i. c. v. injection with or without U0126 changed the general locomotor behaviour, and whether tanshinone I alone or combined with diazepam or MK 801 changed general locomotor behaviour. Briey, the mice were placed in the centre of a horizontal locomotor activity box, and their locomotor activity was measured for 10 min using the video based Ethovision System. All tests were conducted 30 min after the last treatment. Horizontal locomotor activity was converted to total ambulatory distance.

After centrifugation at 18 000 g for 15 min at 4 C, supernatants were subjected to sodium dodecyl sulphate?polyacrylamide gel electrophoresis. Proteins were loaded and size separated by 8?10% SDS?PAGE, and gels were IEM 1754 processed for antigens and blotted onto polyvinylidene diuoride membranes for 1 h. Blots were blocked with Tris buered saline containing 5% non fat dry milk and 0. 01% Tween 20, incubated with anti pERK, anti ERK, anti pCREB, anti CREB or anti BDNF antibodies, and then with secondary antibody conjugated to horseradish peroxidase. Blots were detected using an ECL detection system.