Escalated Oxycodone Self-Administration and Punishment: Differential Expression of Opioid Receptors and Immediate Early Genes in the Rat Dorsal Striatum and Prefrontal Cortex

Abstract

Opioid use disorder (OUD) is characterized by compulsive drug taking despite adverse life consequences. Here, we sought to identify neurobiological consequences associated with the behavioral effects of contingent footshocks administered after escalation of oxycodone self-administration. To reach these goals, Sprague-Dawley rats were trained to self-administer oxycodone for 4 weeks and were then exposed to contingent electric footshocks. This paradigm helped to dichotomize rats into two distinct behavioral phenotypes: (1) those that reduce lever pressing (shock-sensitive) and (2) others that continue lever pressing (shock-resistant) for oxycodone during contingent punishment. The rats were euthanized at 2-h after the last oxycodone plus footshock session. The dorsal striata and prefrontal cortices were dissected for use in western blot and RT-qPCR analyses. All oxycodone self-administration rats showed significant decreased expression of Mu and Kappa opioid receptor proteins only in the dorsal striatum. However, expression of Delta opioid receptor protein was decreased in both brain regions. RT-qPCR analyses documented significant decreases in the expression of c-fos, fosB, fra2, junB, egr1, and egr2 mRNAs in the dorsal striatum (but not in PFC) of the shock-sensitive rats. In the PFC, junD expression was reduced in both phenotypes. However, egr3 mRNA expression was increased in the PFC of only shock-resistant rats. These results reveal that, similar to psychostimulants and alcohol, footshocks can dichotomize rats that escalated their intake of oxycodone into two distinct behavioral phenotypes. These animals also show significant differences in the mRNA expression of immediate early genes, mainly, in the dorsal striatum. The increases in PFC egr3 expression in the shock-resistant rats suggest that Egr3 might be involved in the persistence of oxycodone-associated memory under aversive conditions. This punishment-driven model may help to identify neurobiological substrates of persistent oxycodone taking and abstinence in the presence of adverse consequences.

Oxycodone self-administration activates the mitogen-activated protein kinase/ mitogen- and stress-activated protein kinase (MAPK-MSK) signaling pathway in the rat dorsal striatum

“…Rats were euthanized during early withdrawal, which is defined as the 2 h time point after cessation of drug self-administration. Dorsal striata tissue was dissected as previously described 26 . In brief, we used stereotaxic coordinates (A/P + 2 to -2 mm bregma, M/L ± 2 to 5 mm, D/V − 3 to − 6 mm) according to the rat atlas 27 and we used the position of anatomical structures (corpus callosum and lateral ventricles) for further accuracy.…”

Oxycodone self-administration activates the mitogen-activated protein kinase/mitogen- and stress-activated protein kinase (MAPK-MSK) signaling pathway in the rat dorsal striatum

“…Rats were euthanized 2 h after training day 20. Dorsal striata tissue was dissected as previously described 26 . In brief, we used stereotaxic coordinates (A/P +2 to −2 mm bregma, M/L ±2 to 5 mm, D/V −3 to −6 mm) according to the rat atlas 27 and we used the position of anatomical structures (corpus callosum and lateral ventricles) for further accuracy.…”

Regulation of GABAA Receptor Subunit Expression in Substance Use Disorders

“…The GABRA4 promoter also contains an early growth response protein (Egr) site that is highly conserved across species [ 42 ], and Egr3 has been shown to regulate the change in α 4 subunit expression following a seizure [ 42 ]. Egr family members, especially Egr1, have been shown to be induced in the striatum following substance use [ 43 , 44 ].…”