Molecular pathways implicated in antidepressant actions exhibits diurnal fluctuations in the prefrontal cortex and hippocampus of adult male and female mice.

Epidemiological and experimental studies have demonstrated a bidirectional relationship between mood disorders and the circadian system, with disrupted circadian rhythms contributing to depressive states, and their restoration playing a key role in antidepressants effects. In this context, we sought to examine whether selected molecular pathways implicated in antidepressant responses exhibit diurnal regulatory patterns. Naïve adult male and female C57BL/6 mice were euthanized at 3-hour intervals beginning at Zeitgeber Time 0 (ZT0), and hippocampal (HC) and medial prefrontal cortex (mPFC) tissues were collected for RT-qPCR and western blot analyses. We observed statistically significant diurnal rhythmicity in all analyzed transcripts (cFos, Arc, Nr4a1, Dusp1, Dusp5, and Dusp6) in both HC and mPFC samples, with peak expression occurring during the dark (active) phase (ZT15-18). Phosphorylation levels of TrkBY816 (tropomyosin-related kinase) and GSK3βS9 (glycogen synthase kinase 3β) also showed patterns suggestive of temporal rhythmicity, peaking during the light (inactive) phase. Levels of p-ERK2T185/Y187 (extracellular-signal regulated kinase) did not display rhythmic fluctuations, but peaked during the light phase in the HC, especially in males. Collectively, these findings demonstrate that antidepressant-related signaling is subject to diurnal regulation, highlighting the importance of integrating circadian biology and time-of-day as relevant variables in the development of translationally relevant antidepressant research.