Morris Water Maze

BEHAVIOURAL PARAMETERS:

Morris water maze

Morris water maze consists of a large circular pool (100 cm in diameter, 35 cm in height), filled to a depth of 30 cm with water. Water was made opaque with titanium dioxide. The pool was divided arbitrarily, into four equal quadrants [12]. A clear Plexiglas platform with a diameter of 11cm was submerged 1cm below the water level. The platform was placed near the centre of a quadrant and rats were released into the water from 1 of the 3 remaining quadrants to search for the platform. The time taken to find the hidden platform [escape latency (EL)] was recorded in each trial. Even after 90 seconds if animal did not reach the platform, it was guided to the platform and allowed to remain on platform for

The supernatant was assayed for SOD activity by following the inhibition of epinephrine auto-oxidation. 0.5ml of sample was diluted with 0.5 ml of distilled water, to this 0.25 ml ethanol, 0.5 ml of chloroform (all reagents chilled) was added. The mixture was shaken for 1 min and centrifuged at 2000 rpm for 20 min. The enzymatic activity in supernatant was determined. To 0.05 ml of carbonate buffer (0.05 M, pH 10.2) and 0.5 ml of EDTA (0.49 M) was added. The reaction was initiated by the addition of 0.4ml of epinephrine and the change in optical density/min was measured at 480nm. SOD activity was expressed as units/mg protein change in optical density/min. 50% inhibition of epinephrine to adrenochrome transition by enzyme is taken the enzyme unit. Calibration curve was prepared by using 10 -125 units of

A total of 0.1 ml of supernatant was added to cuvette containing 1.9 ml of 50mM phosphate buffer (pH 7). The reaction was started by the addition of 1 ml freshly prepared 30mM H2O2. The rate of decomposition of H2O2 was measured spectrophotometrically at 240 nm. Catalase values were expressed as n moles H2O2 consumed/min/mg protein.

Measurement of lipid peroxidation

TBARS, a measure of lipid per oxidation, was measured as described by Ohkawa [15]. Briefly, 1 ml of suspension medium was taken from the 10% tissue homogenate. 0.5 ml of 30% Trichloroacetic acid (TCA) was added to it, followed by 0.5 ml of 0.8% thiobarbituric acid (TBA) reagent. The tubes were covered with aluminium foil and kept in shaking water bath for 30 minutes at 80°C. After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes. These were centrifuged at 3000 rpm for 15 minutes.

The absorbance of the supernatant was read at 540 nm at room temperature against appropriate blank. Blank consist of 1 ml distilled water, 0.5 ml of 30% TCA, and 0.5 ml of 0.8% TBA. TBARS values were expressed as n moles malonaldehyde (MDA)/mg protein.

Estimation of