Ed atop the platform.

The results were analyzed as the percentage

Ed atop the platform. The results were analyzed as the percentage of correct choices, using repeated-measures ANOVA.Elevated plus-maze testA three-arm, walled T-maze,constructed of white Plexiglas (600 mm along the stem, 800 mm side at the T-intersection, 400 mm high, with passages 100 mm wide), was situated in one corner of a brightly lit behavioral-testing room separate from the colony. The T-maze was refilled daily with 145 mm of water at 2 so that a platform (140 mm high, 300 mm2 in size), rising from the floor of the maze, was submerged 5 mm below the water line. One day prior to initial training, mice were placed in the maze and allowed to swim for 60 seconds with no platform present. The platform was then inserted in a standardized position 80 mm from the end of a goal arm, and each mouse was placed directly on the platform for 30 seconds. Finally, each mouse was placed at the far end of the stem and allowed to locate the submerged goal-arm platform. On each of four consecutive training days, a forced-choice alternation paradigm required each subject to perform eight replications PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/11834444 of a paired forced-choice/free-choice trial sequence. With either the left or PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/8086425 right goal arm blocked with a guillotine door, Methyl 5-amino-2,4-difluorobenzoate each subject was placed in the far end of the stem, and allowed to ascend the submerged platform located in the goal arm opposite the blocked arm. The animal remained atop the platform for 15 seconds at the conclusion of this forced choice trial. The animal was then removed by the tail and again placed at the end of the stem, while simultaneously,The elevated plus maze was made from polyvinylchloride, and built in the shape of a plus sign, with two open (white) arms (340 ?75 ?10 mm) and two closed (black) arms (340 ?75 ?175 mm) opposite each other. The center of the four arms comprised the middle square (75 ?75 mm). The maze was elevated 510 mm above ground level. Each mouse was placed separately in the centre of the maze, facing an open arm, and allowed to 4-(Benzyloxy)-4-oxobutanoic acid explore the apparatus freely for 5 minutes. Parameters measured included the number of entries into the closed and open arms (an index of motor function), and the length of time spent in the closed and open arms. An entry was counted only after the mouse entered the arm with four paws. Before each test, the box was cleaned with a diluted alcohol solution to eliminate smells. The percentage of entries into the open arms out of the total number of arm entries and the percentage of time spent in the open arms, which are all accepted measures of anxiety levels, were further calculated.Histological studiesMice were anesthetized by intraperitoneal injection of ketamine (100 mg/kg) and xylazine (20 mg/kg) and underwent transcardiac perfusion with phosphate buffer saline followed by perfusion with 4 paraformaldehyde in PBS. Brain tissue was collected, fixed in 4 paraformaldehyde and embedded in paraffin wax. Coronal sections 6 m thick were cut, mounted, and stained with hematoxylin and eosin (H E), Luxol Fast Blue (LFB), and Bielchowsky (BLS) stains to identify histological details and the density of myelin and axons, and the sections were specifically examined to evaluate ischemic pathology such as microinfarcts.Katzav et al. BMC Medicine 2013, 11:92 http://www.biomedcentral.com/1741-7015/11/Page 4 ofImmunohistochemistryParaffin wax-embedded sections were dewaxed and rehydrated in xylene and alcohol solutions, 4-dione 4-Bromo-3-hydroxypyridine Dibenzo[b then rinsed with PBS. Citrate buffer was used for antigen retrieval, and.