Experimental Design.(A) For the gene expression qRT-PCR study, triplet rats were assigned to control, chronic exposure, and four- or forty-eight-hour withdrawal. Following a 35-day liquid alcohol exposure, animals were sacrificed and the RNA extracted from their CeA and DVC. (B) For IHC study, four pairs of animals were assigned to either the chronic exposure or 48 hours of alcohol withdrawal following eight months on the alcohol diet. Forty-eight hours prior to the time of sacrifice, the withdrawal animals had their diets removed to initiate withdrawal. Following intracardiac perfusion, both rats were sacrificed and their CeA and DVC collected for immunohistochemistry and confocal microscopy. CeA, central nucleus of the amygdala; DVC, dorsal vagal complex; qRT-PCR, quantitative reverse transcription polymerase chain reaction; IHC, immunohistochemistry.
Freeman et al. Journal of Neuroinflammation 2012 9:97 doi:10.1186/1742-2094-9-97