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p.) and perfused intracardially with saline solution. Brains were removed and stained by modified Golgi-Cox method as described previously (Alcantara-Gonzalez et al., 2010; Bringas et al., 2012; Flores et al., 2005; Gibb and Kolb, 1998; Juarez et al., 2008). Coronal sections of 200-?m thickness from the PFC, hippocampus and BLA were obtained using a vibratome (Campden Instruments, MA752, Leicester, UK). These sections were collected on clean gelatin-coated microscope slides and treated with ammonium hydroxide for 30 min, followed by 30 min in Kodak film fixer and subsequently washed with distilled water, dehydrated and cleared in successive baths of 50% (1 min), 70% (1 min), 95% (1 min), Oxygenase and 100% (2 �� 5 min) ethanol followed by 15 min in a xylene solution. Subsequently, the slides were covered with balsam resinous medium. The cortical thickness (in mm) was measured in the Golgi-Cox-stained slides using a Zeiss-Jena projector at a magnification of X 10 (Mychasiuk et al., 2012). Three PFC subregions were analyzed, Cg1, prelimbic and infralimbic areas, which were measured for four different plates (3.7, 3.2, 2.7, and 2.2 mm from Bregma; Paxinos and Watson, 1998) in each hemisphere. The CA1 area of the dorsal hippocampus was analyzed. This region was measured for six different plates (?2.6, ?3.14, ?3.3, ?3.6, ?3.8, and 4.16 mm from Bregma; Paxinos and Watson, 1998). Finally, the area of BLA was measured for nine different plates (?1.6, ?1.8, ?1.88, ?2.12, ?2.3, ?2.56, ?2.8, ?3.14, and ?3.3 mm from Bregma; Paxinos and Watson, 1998) by using a software developed by BUAP (with the help of the software MATLAB this website Versi��n 5), using which the separate plates of each region were collapsed to get a representative analysis of each cortical Fasudil concentration thickness region and the area of the total BLA (Fig. 1). The mean values for each brain region were treated as a single measurement for the data analysis (n?=?7�C9). The data were analyzed by two-way ANOVA, followed by the Sidak test for post hoc comparisons, with prenatal VAP exposure and age as independent factors. P?
