ANALYSIS OF THE DORSAL RAFE NUCLEUS TRANSCRIPTOME AFTER CHALLENGES TO HYDROELETROLYTIC AND ENERGY HOMEOSTASIS
Transcriptome, Sodium appetite, Food deprivation
The sodium ion is as a key element in the regulation of the hydroelectrolytic balance, so that its levels must be kept in a narrow range compatible with life. For such regulation, a series of renal, endocrine and behavioural mechanisms are activated. The dorsal raphe nucleus (DRN) has been implicated as one of the main brain nuclei involved in the modulation of sodium-specific appetite. In this work we investigate the possible molecular mechanisms involved in modulating sodium appetite through the development of transcriptomes. Male Wistar rats, approximately 60 days old, were randomly separated into 4 main experimental groups: a) control (CTRL); b) low sodium diet (LS); c) furosemide (FURO); d) salt loading (SL); and an additional group that was subjected to food deprivation for 48 hours. After the experimental period, euthanasia was performed to collect blood and brain. Animals submitted to treatment with furosemide and saline overload showed an increase in haematocrit (FURO 45.6 ± 6.8; SL 46.6 ± 5.2 vs. CTRL 40.3 ± 3.4) and changes in osmolality (FURO 264.8 ± 20; SL 296.6 ± 23 vs. CTRL 278.3 ± 11.7). The DRN RNA sequencing resulted in approximately 18,600 genes mapped in total. Among the most expressed genes, tryptophan hydroxylase isoform 2 (Tph2), the gene encoding the serotonin transporter SERT (Slc6a4) and the vesicular monoamine transporter (Slc18a2) were found, confirming the prevalence of serotonergic neurons. The comparison of differentially expressed genes using the Venn diagram revealed a low altered number in the FURO group (6 genes), no altered genes in the DP group and 22 genes altered by saline overload. Based on the results, we performed the PCR validation only for the saline overload group. Among the 22 altered genes in the transcriptome, we selected 10 targets for PCR, among which, 7 genes were successfully validated. Finally, the sequencing of the group subjected to food deprivation revealed 111 altered genes, when comparing this result to that obtained in salt loading, we saw that the groups have 6 altered genes in common. Despite the need to validate the results of food deprivation, they indicate a possible overlap in the level of DRN between mechanisms that regulate specific sodium appetite and eating behaviour.