Transcriptomic modulation of CeA and BnST following gastrointestinal vagal deafferentation in male rats

The aim of the study was to investigate the effects of gastrointestinal vagal deafferentation on gene expression in two brain areas related to anxiety-like behavior (the central nucleus of the amygdala CeA and the bed nucleus of the stria terminalis). For this, male Sprague-Dawley rats (n=8, 9 weeks of age) were injected blaterally into the nodose ganglia with saporin (SAP) or cholecystokinin cojugated with SAP (CCK-SAP) to induce gastrointestinal vagal deafferentation as previously described (Diepenbroek et al., Am J Physiol, 2015). After a battery of tests investigating anxiety-like behavior (open field, elevated plus maze, acoustic startle reflex and food neophobia) and a verification of the functionality of the neuronal lesion (attentuated anorexigenic response to ip CCK), rats were sacrificed at 30 weeks of age. After anesthesia, rats were perfused for 3 min with oxygenated aCSF and their brains were immediately extracted and frozen in ice cold isopentane. Brain micropunches of CeA and BnST were collected in a cryostat and total RNA extracted with Trizol. Before RNA sequencing, the integrity of mRNA from brain micropunches was verified using the Agilent 2100 bioanalyzer. Samples with RNA integrity number (RIN) above 8 were further processed and included in the analysis. RNA sequencing was performed by Novogene.