Fentanyl exposure alters rat CB1 receptor expression in the insula, nucleus accumbens and substantia nigra

Prolonged periods of opioid use have been shown to cause neuroadaptations in the brain's reward circuitry, contributing to addictive behaviors and drug dependence. Recently, considerable focus has been placed on the role of the endocannabinoid system (ECS) and its CB receptors in opioid-driven behaviors. However, opioid-induced neuroadaptations to the ECS remain understudied. In this study, we systematically assessed CB1 receptor (CB1R) protein expression within the cortico-mesolimbic-basal ganglia circuit in rats following chronic fentanyl exposure. Male and female Long Evans rats were administered increasing daily doses of fentanyl or saline for 14 days. During naloxone-precipitated withdrawal, fentanyl-treated rats exhibited significantly higher withdrawal symptoms than saline-treated controls. Using Western Blotting, we demonstrated that the fentanyl-treated rats had significantly higher CB1R expression in the insula and significantly lower CB1R expression in the nucleus accumbens and substantia nigra compared to saline-treated rats. No significant differences in CB1R expression were detected between saline and fentanyl-treated rats in the prefrontal cortex, dorsal striatum, medial septum, hypothalamus, amygdala, hippocampus, ventral tegmental area, periaqueductal gray area, pedunculopontine tegmentum, and laterodorsal tegmentum (LDT). These findings suggest that chronic fentanyl exposure leads to region-specific neuroadaptations of CB1R protein expression in motivation- and addiction-associated brain regions.