Transcriptome analysis of the bed nucleus of the stria terminalis (BNST) of mice subjected to the rat exposition stress-induced reinstatement of ethanol seeking

• Grant number: 22/09953-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: September 01, 2022
• End date: November 30, 2024
• Field of knowledge: Biological Sciences - Pharmacology - Neuropsychopharmacology
• Principal Investigator: Fabio Cardoso Cruz
• Grantee: Ben Tagami Rodolpho
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated research grant: 18/15505-4 - Neurobiology study of relapse to alcohol and cocaine seeking: identification of plasticity in neuronal ensembles that encodes addiction-related memories, AP.JP2
• Associated scholarship(s): 23/17928-8 - Analysis of hevin function on ethanol-induced plasticity in dopamine hub synapses in the amygdala., BE.EP.MS

Abstract

Ethanol dependence is a multifactorial disorder influenced by biopsychosocial variants. In this context, exposure to stressful situations during abstinence can increase the individual's vulnerability to relapse to using this substance even after long periods. Although relevant, the neurobiology of the interaction between stress and relapse to ethanol use is still not fully understood. Thus, the present study aims to evaluate whether the exposure of mice to the Rat Exposure Test may promote the reinstatement of ethanol-seeking behavior. Adolescent male mice will be isolated (stress group) or kept grouped (control group) from the post-natal day (PND)28. On PND 60, they will have access to a bottle containing 9% ethanol + 2% sucrose or 2% sucrose for 4 hours daily for seven days in their home cage. Subsequently, they will be trained for operant ethanol self-administration. After the acquisition of the self-administration, the animals will undergo extinction of this behavior, in which responses to the active nose poke will not be reinforced. Finally, the stress-induced reinstatement of ethanol-seeking will be tested by exposing mice to the rat exposure protocol. After the reinstatement test, the animals will be perfused, and the brains will be collected for immunofluorescence analysis for the proteins Fos, CamkII, GAD67, and NeuN, in the interstitial nucleus of the stria terminalis (BNST), in order to characterize the neuronal ensembles present in this structure. In the second experiment, the brains will be collected and processed immediately after the reinstatement test to perform the BNST transcriptome analysis. In the third experiment, after the test, the brains will be collected and processed for gene expression analysis only in neuronal ensembles of the BNST, using flow cytometry for activated cells followed by RT-PCR.