In the adult mammalian brain, new neurons continue to be produced

In the adult mammalian brain, new neurons continue to be produced throughout life in two main regions in the brain, the subgranular zone (SGZ) in the hippocampus and the subventricular zone in the walls of the lateral ventricles. cell proliferation, migration and differentiation in the dentate Brivanib gyrus (DG) of the hippocampus of adult mice were analyzed using specific markers. Cognitive recovery was evaluated by screening the animals for acknowledgement, spatial and associative learning and memory. Cognitive function was maintained by CAST overexpression following seizures, while modulation of post-injury neurogenesis was comparable to wild type (WT) mice. Calpain inhibition could still be potentially able to prevent the impairment in the formation of new neurons, given that the levels of calpain activity could be WASF1 reduced under a certain threshold and other harmful effects from the pathological environment could also be controlled. in a 12-h dark:light cycle. All experiments were performed in accordance with institutional and European guidelines (2010/63/EU) for the care and use of laboratory animals. The procedures performed in mice explained in this work have been examined and approved by the Animal Welfare Body of the Center for Neuroscience and Cell Biology and have been approved by the Direc??o Geral de Alimenta??o at the Veterinria (reference 0421/000/000/2013). KA Treatment Hippocampal damage was induced by administering KA subcutaneously (25 mg/kg, in a concentration of 5 mg/ml, in a saline (SAL) answer of 0.9% NaCl), as previously explained by our group (Carreira et al., 2010). After KA administration, the animals went through several stages, according to a well-defined level (Schauwecker and Steward, 1997): immobility (I), tail/forelimb extension/rigid posture (II), repeated movements/head bobbing (III), rearing and falling (IV), continuous rearing and falling (V), severe tonic-clonic seizures (VI). Only mice that reached stage V or higher were used in this study. SAL-treated animals were used as controls. Behavior Analysis and Neuronal Differentiation For the studies of short-term behavioral recovery, three different behavioral assessments were performed Brivanib in WT and hCAST mice, on the 3rdeb week after KA or SAL treatment (Physique ?(Figure1A):1A): object recognition (OR; Physique ?Determine1C),1C), water maze (WM; Physique ?Physique1Deb)1D) and fear conditioning (FC; Physique ?Physique1E),1E), as explained further in more detail. To assess long-term behavioral recovery and neuronal differentiation after lesion, WT and hCAST mice were treated as shown in Physique ?Figure1B.1B. The thymidine analog 5-ethynyl-2-deoxyuridine (EdU) was intraperitoneally given to WT and hCAST mice on days 3, 4 and 5 after KA or SAL treatment, and the animals were sacrificed by transcardial perfusion after 8 weeks, when new neurons are fully mature (Aimone et al., 2014). Behavioral analysis was performed during the last week. Physique 1 Experimental procedures for the study of behavioral recovery and neurogenesis after Brivanib kainic acid (KA) or saline (SAL) treatment. Wild type (WT) and hCAST mice were treated with either SAL or KA (25 mg/kg, s.c.). (A) To study behavioral recovery in the … Object Acknowledgement For the study of acknowledgement memory, the OR test (Bevins and Besheer, 2006) was used. Mice were offered to two comparable objects, following habituation to the apparatus (40 40 40 cm3 acrylic box), after which one of the objects was replaced by a novel one. Each stage of the test experienced the duration of 5 min, 2 h apart, and the apparatus and objects were washed with 10% ethanol between animals. In the presence of the objects, the percentage of time spent with each object (totalizing 100%) was assessed. The analysis was carried out in videos acquired with a GoPro Hero (GoPro, Inc., San Mateo, CA, USA) during screening of 3C5 animals per group (for short-term recovery) and 12C15 animals per group (for long-term recovery), using the Any-maze software (version 4.99, Stoelting Co., Solid wood Dale, IL, USA). Morris Water Maze To evaluate spatial learning and memory, the WM test (Morris et al.,.