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Fig. 1 | Molecular Autism

Fig. 1

From: Hippocampal neurons isolated from rats subjected to the valproic acid model mimic in vivo synaptic pattern: evidence of neuronal priming during early development in autism spectrum disorders

Fig. 1

Experimental design. a Prenatal VPA injection defined the experimental groups. At E10.5, either VPA (450 mg/kg, i.p) or saline were intraperitoneally administrated to pregnant rats, and the corresponding pups were assigned to VPA or control groups, respectively. Postnatal day (PND) 0 was designated the day pups were born. b Early postnatal and juvenile VPA behavioral deficits were determined by the evaluation of the swimming performance (PND8-14) and exploration and social interaction in control and VPA animals (PND30-35). At PND3 and PND35, brains from control and VPA rats were processed for immunofluorescence (IF) assays or transmission electron microscopy (TEM) studies. The in vitro studies involved the dissection of the hippocampus from control and VPA animals to establish primary neuronal cultures (PND1) and for mixed glial primary cultures (PND3). Hippocampal neurons were fixed for immunofluorescence (IF) assays at day in vitro (DIV) 7, 10 and 14. Also, at DIV14, the FM4-64 assay was performed, and neurons were obtained for western blot (WB). Besides, a brief stimulus with glutamate was used to study structural synaptic remodeling at DIV14. In the case of glial cultures, microglial cells were obtained from mixed glial cultures. After 7 DIV, cell size and internal complexity were evaluated by flow cytometry and their morphological response to different stimuli [LPS and synaptosomes (ST)] was analyze by IF assays

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