Elsevier

Biological Psychiatry

Volume 84, Issue 12, 15 December 2018, Pages 881-892
Biological Psychiatry

Archival Report
E3 Ubiquitin-Protein Ligase SMURF1 in the Nucleus Accumbens Mediates Cocaine Seeking

https://doi.org/10.1016/j.biopsych.2018.07.013Get rights and content

Abstract

Background

Substance use disorder is a neurobiological disease characterized by episodes of relapse despite periods of withdrawal. It is thought that neuroadaptations in discrete brain areas of the reward pathway, including the nucleus accumbens, underlie these aberrant behaviors. The ubiquitin–proteasome system degrades proteins and has been shown to be involved in cocaine-induced plasticity, but the role of E3 ubiquitin ligases, which conjugate ubiquitin to substrates, is unknown. Here, we examined E3 ubiquitin-protein ligase SMURF1 (SMURF1) in neuroadaptations and relapse behavior during withdrawal following cocaine self-administration.

Methods

SMURF1 and downstream targets ras homolog gene family, member A (RhoA), SMAD1/5, and Runt-related transcript factor 2 were examined using Western blotting (n = 9–11/group), quantitative polymerase chain reaction (n = 6–9/group), co-immunoprecipitation (n = 9–11/group), tandem ubiquitin binding entities affinity purification (n = 5–6/group), and quantitative chromatin immunoprecipitation (n = 3–6/group) (2 rats/sample). Viral-mediated gene transfer (n = 7–12/group) and intra-accumbal microinjections (n = 9–10/group) were used to examine causal roles of SMURF1 and substrate RhoA, respectively, in cue-induced cocaine seeking.

Results

SMURF1 protein expression was decreased, while SMURF1 substrates RhoA and SMAD1/5 were increased, in the nucleus accumbens on withdrawal day 7, but not on withdrawal day 1, following cocaine self-administration. Viral-mediated gene transfer of Smurf1 or constitutive activation of RhoA attenuated cue-induced cocaine seeking, while catalytically inactive Smurf1 enhanced cocaine seeking. Furthermore, SMURF1-regulated, SMAD1/5-associated transcription factor Runt-related transcript factor 2 displayed increased binding at promoter regions of genes previously associated with cocaine-induced plasticity.

Conclusions

SMURF1 is a key mediator of neuroadaptations in the nucleus accumbens following cocaine exposure and mediates cue-induced cocaine seeking during withdrawal.

Section snippets

Subjects

Adult male Sprague Dawley rats (250–275 g; Envigo, Indianapolis, IN) were singly housed on a reverse light/dark cycle and had ad libitum access to food and water. This study was conducted in accordance with the guidelines set up by the Institutional Animal Care and Use Committee of The State University of New York at Buffalo.

Drugs

Cocaine hydrochloride, gifted by the National Institute on Drug Abuse (Bethesda, MD), was dissolved in sterile 0.9% saline (4.5 mg/mL). Pump durations/injection volumes

SMURF1 Is Decreased in the NAc in a Withdrawal-Dependent Manner Following Cocaine Self-administration

RhoA and the TGF-β superfamily have previously been implicated in cocaine-dependent plasticity 22, 23, 24, 25, 26, 32, 33. We therefore set out to determine whether SMURF1, which regulates RhoA and BMP signal transducers SMAD1 and SMAD5, is involved in cocaine-related cellular and behavioral plasticity. To do so, we first asked whether SMURF1 and substrates are altered after cocaine exposure. Rats were trained to nose poke to receive intravenous infusions of saline or cocaine paired with a

Discussion

Long-term neuroadaptations in brain regions that process reward and motivation, including the NAc, underlie relapse vulnerability. The UPS governs activity-dependent synaptic plasticity 5, 6, 64, 65 and has been shown to be involved in cocaine-related plasticity in the NAc 15, 16, 66, 67. However, E3s, which mediate polyubiquitination of substrates for proteasomal degradation, have not been examined in substance use disorder. In this study, we examined the E3 SMURF1 in cellular and behavioral

Acknowledgments and Disclosures

This work was supported by the National Institutes of Health (National Institute on Drug Abuse [NIDA] Grant No. R01DA037257 [to DMD], Grant No. S1-R01DA037257 [to DMD], NIDA Grant No. R21DA044486 [to DMD], National Institute of Neurological Disorders and Stroke Grant No. F99NS108543 [to JAM], and National Institute of General Medical Sciences Grant No. R25GM09545902 [to University at Buffalo]). The NIDA Drug Supply Program generously gifted the cocaine used in these studies.

CTW, RV, J-XL, and

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    CTW and RV contributed equally to this work.

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