
==== Front
Mol Psychiatry
Mol Psychiatry
Molecular Psychiatry
1359-4184
1476-5578
Nature Publishing Group UK London

38351172
2455
10.1038/s41380-024-02455-2
Article
Incubation of methamphetamine craving in punishment-resistant individuals is associated with activation of specific gene networks in the rat dorsal striatum
Daiwile Atul P.
McCoy Michael T.
Ladenheim Bruce
Subramaniam Jayanthi
http://orcid.org/0000-0001-5635-3524
Cadet Jean Lud jcadet@intra.nida.nih.gov

grid.94365.3d 0000 0001 2297 5165 Molecular Neuropsychiatry Research Branch, NIH/NIDA Intramural Research Program, Baltimore, MD 21224 USA
14 2 2024
14 2 2024
2024
29 7 19902000
3 7 2023
18 1 2024
23 1 2024
© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2024
2024
https://creativecommons.org/licenses/by/4.0/ Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
Methamphetamine use disorder (MUD) is characterized by loss of control over compulsive drug use. Here, we used a self-administration (SA) model to investigate transcriptional changes associated with the development of early and late compulsivity during contingent footshocks. Punishment initially separated methamphetamine taking rats into always shock-resistant (ASR) rats that continued active lever pressing and shock-sensitive (SS) rats that reduced their lever pressing. At the end of the punishment phase, rats underwent 15 days of forced abstinence at the end of which they were re-introduced to the SA paradigm followed by SA plus contingent shocks. Interestingly, 36 percent of the initial SS rats developed delayed shock-resistance (DSR). Of translational relevance, ASR rats showed more incubation of methamphetamine craving than DSR and always sensitive (AS) rats. RNA sequencing revealed increased striatal Rab37 and Dipk2b mRNA levels that correlated with incubation of methamphetamine craving. Interestingly, Bdnf mRNA levels showed HDAC2-dependent decreased expression in the AS rats. The present SA paradigm should help to elucidate the molecular substrates of early and late addiction-like behaviors.

Subject terms

Neuroscience
Molecular biology
Psychology
https://doi.org/10.13039/100000026 U.S. Department of Health & Human Services | NIH | National Institute on Drug Abuse (NIDA) DA000552 (2021) Cadet Jean Lud issue-copyright-statement© Springer Nature Limited 2024
==== Body
pmcIntroduction

Methamphetamine (METH) is a highly addictive psychostimulant with a very high prevalence of misuse throughout the world [1]. Many, but not all, METH users meet diagnostic criteria for METH use disorder (MUD) [2–4]. These criteria include excessive drug taking during binges and compulsive METH use despite adverse consequences [2]. Compulsive use is thought to be a fundamental feature of addictive diatheses [5–7]. However, there is no FDA-approved medication to treat individuals with MUD. The development of pharmacological therapeutics against MUD might depend on the elucidation of METH-induced molecular and biochemical changes that occur in various cortical and subcortical brain regions that subserve reward, decision-making, and habit forming [8–16]. The fact that only a subset of METH users meets criteria for MUD [2–4], suggests the presence of differences in the molecular substrates in the reward circuitries that control the development of MUD in various individuals. A brain region of special interest is the dorsal striatum, which serves as a key regulator of habit forming [17] and participates in the development and progression of compulsive drug taking in models of substance use disorders (SUDs) [18–23]. The dorsal striatum might also serve as a node in neuronal pathways involved in the tenacity of misuse of licit and illicit substances despite nefarious consequences associated with their abuse [24]. The role for the dorsal striatum in METH-induced behavioral consequences is supported by observations of epigenetic and transcriptional changes in the dorsal striatum of rats that had self-administered the drug compulsively over several weeks [10, 25–27].

In order to clarify the role of striatal molecular mechanisms in compulsive METH taking, we have conducted studies in rats that meet the DSM5 criterion of compulsive drug taking in the presence of adverse consequences represented by contingent footshocks during METH self-administration (SA) experiments [25, 28–32]. We have reported that rats which continue to take METH compulsively in the presence of footshocks showed greater incubation of METH craving than animals that had suppressed their METH intake [13, 31, 33]. We also found that compulsive SA behaviors are also correlated with differences in the balance between orbitofrontal and prelimbic striatal circuits of rats [30], findings that are clinically relevant in view of the relevance of these circuits in SUDs [34–36]. Moreover, there was differential expression of genes and proteins in the dorsal striatum of compulsive and non-compulsive rats [13, 37], potentially implicating that structure in the behavioral differences observed in these two METH SA phenotypes.

The present study was undertaken to investigate the time course of punishment sensitivity over several weeks and to identify genes and molecular pathways associated with these behaviors by using a genome-wide gene expression platform. Several groups of investigators have used genome-wide approaches to identify genes of interest in animal models of cocaine [38, 39], heroin [16], and morphine [40] use, as well as in the case of psychiatric diseases [41]. Herein, we provide convincing evidence for the existence of rats that develop persistent punishment-resistant METH SA over several weeks. We also report that a subpopulation of rats that had initially suppressed their drug intake in the presence of shocks developed delayed shock resistance (DSR) after 2 weeks of forced abstinence. Global analysis of gene expression by RNA sequencing revealed the presence of many differentially expressed genes (DEGs) in the dorsal striatum of compulsive (addicted) versus non-compulsive (non-addicted) rats. Finally, chromatin immunoprecipitation studies identified HDAC2 as an upstream epigenetic regulator of the expression of some trophic factors including Bdnf that had shown decreased expression in the non-compulsive rats.

Materials and methods

Animals and intravenous surgery

Male Long Evans rats weighing 350–400 g were purchased from Charles River, USA. They were group-housed with free access to food and water. We performed intravenous surgery as described in our previous publications [28, 42, 43]. More details are provided in supplementary text section 1. All animal procedures were approved by the National Institute of Drug Abuse Animal Care and Use Committee (Protocol No. 21-MNPB-10) and conducted according to the Guide for the Care and Use of Laboratory Animals (ISBN 0-309-05377-3).

METH self-administration

METH dose (0.1 mg/kg/infusion), METH SA training procedures (three 3-h sessions/day separated by a 30-min off interval between each session) for 20 days under a fixed-ratio-1 (FR-1) schedule, footshock phase, and METH seeking tests on withdrawal day 1 (WD1) and 15 (WD15) were performed according to published protocols [13, 28, 44]. As described previously 50% of the reinforced active lever presses resulted in the simultaneous delivery of a 0.5-s footshock through the grid floor [28]. The always shock-resistant (ASR) and shock-sensitive (SS) rats were separated as per our previous published studies [28, 33]. The detailed METH SA training procedures are described in supplementary text section 2.

At the end of the SA plus contingent sessions, rats underwent METH seeking tests at withdrawal days 1 and 15 (see Fig. 1A). Following these tests, rats were placed back inside their respective SA chambers for a second METH SA training phase that lasted for 12 days (Fig. 1A). During the last 3 days of the second METH training phase, rats were again exposed to contingent footshocks at an intensity of 0.30 mA. Subsequently, rats were removed from the SA boxes and housed individually in the animal vivarium with no access to METH. They then underwent a second set of cue-induced METH seeking on withdrawal days 1 (WD1) and 15 (WD15) in their respective SA chambers.Fig. 1 Extended METH self-administration (SA) and contingent footshocks result in compulsive METH taking in a subpopulation of rats.

A Experimental timeline showing long access METH SA sessions and contingent footshock sessions. Footshocks were administered randomly during 50% of pressing the active lever for METH. B During the first phase of the experiment, footshocks reduced lever pressing in shock-sensitive (SS, n = 14) but not in always shock-resistant (ASR, n = 12) or compulsive rats. Always shock-resistant (ASR) and shock-sensitive (SS) rats were separated as per our previous published study in which we classified animals as shock-sensitive if they reduced their intake by 60% [33]. C Resistant/compulsive rats showed greater incubation of METH craving than SS rats at withdrawal day 15 (WD15) during forced abstinence. D SS rats were separated post-facto during the second phase of METH SA training because a subset of the SS rats, now labeled ‘Delayed shock resistance (DSR)’, developed shock-resistance during the second phase of the behavioral experiments. We named the remaining SS rats ‘Always sensitive (AS)’ (see H). E Footshocks caused marked reduction in METH intake in the sensitive rats during the first phase of footshocks. The figure shows METH intake during the last 3 days of training without shocks (L 3 d) and the last 3 days of the first footshock phase (FS L 3 d). F ASR rats showed greater incubation of METH craving in comparison to AS rats but not DSR rats during tests of relapse during the first forced abstinence period. G Regression analysis shows a positive correlation between METH intake during first sets of footshock days and active lever responding during the relapse test on WD15 of the first phase of the behavioral experiments. H Footshocks reduced lever pressing for METH in the AS rats but not in the ASR and DSR rats. I Footshocks reduced METH intake in the AS but not in the ASR and DSR rats. J Persistently compulsive SR (ASR) rats continued to show incubation of METH craving in comparison to AS rats during the second set of relapse tests. K Regression analysis showed a positive correlation between METH intake during footshock days and active lever responding on the second WD15 relapse test. CT, saline; ASR, always shock-resistant; SS, shock-sensitive, DSR, delayed shock-resistant; and AS, always sensitive rats. Key to statistics: *p < 0.05, **p < 0.01, ***p < 0.001, comparisons between METH groups (ASR, DSR, AS) and controls; #p < 0.05, ##p < 0.01, ###p < 0.001, comparisons between ASR and AS or SS; ! p < 0.05, !!p < 0.01, !!!p < 0.001, comparison between ASR and DSR; $p < 0.05, $$p < 0.01, $$$p < 0.001, comparison between DSR and AS; &p < 0.05, &&p < 0.01, &&&p < 0.001, comparison between before vs after footshocks; @p < 0.05, @@p < 0.01, @@@p < 0.001, comparison between WD1 and WD15.

Tissue collection

Rats were euthanized 24 h after the drug seeking test on WD15 by rapid decapitation with a guillotine. Using specific neuroanatomical coordinates obtained a rat atlas, dorsal striata (dSTR, A/P + 2 to −2 mm bregma, M/L ± 2 to 5 mm, D/V −3 to −6 mm) were dissected out and immediately snap-frozen on dry ice and stored at −80 °C.

RNA Sequencing

Total RNA was isolated from the dSTR using Qiagen RNeasy Mini kit (Qiagen, USA) and quantified with nanodrop. RNA integrity (RIN) was checked using the Agilent bioanalyzer 2100. RNA samples with RIN 8 or above were shipped on dry ice to Azenta, Genewiz (USA) for RNA sequencing (More details are provided in supplementary text section 3). RNA sequencing data have been deposited at the NCBI under the accession # GSE220896.

Quantitative polymerase chain reaction (qPCR)

We used qPCR to validate the changes in expression of several genes of interest identified as differentially expressed in the RNA-Seq analysis. Advantage RT-for-PCR kit (Clontech) was used to reverse transcribed 500 ng of total RNA using oligo dT primers. Using iQ SYBR Green Supermix (Bio-Rad, USA), qPCR was performed with Roche LightCycler 480 II system. The relative mRNA expression was normalized to beta-2-microglobulin (B2M). The qPCR primer sequences used in the study are listed in supplementary Table 1.

Chromatin immunoprecipitation (ChIP) and PCR

Chromatin immunoprecipitation using an antibody specific for HDAC2 was carried out as per our previously published protocol [45]. More details are also provided in supplementary text section 4. Real-time qPCR was also performed with the Roche LightCycler 480 II system using promotor specific ChIP-PCR primers listed in Supplementary Table 1. The primers were procured from the Synthesis and Sequencing Facility of Johns Hopkins University.

Statistical Analyses

Behavioral mRNA and ChIP-PCR data were analyzed with the statistical program GraphPad Prism 9 using ANOVA with repeated measures and followed by Fishers protected least significant difference (PLSD) test. The detailed statistical analyses are described in supplementary text section 5.

Results

A subpopulation of rats exhibits compulsive METH taking despite punishment

Figure 1A illustrates the timeline of our behavioral experiment. We analyzed the behavioral data using repeated measures two-way ANOVA with group (saline vs METH) and training days (22 days) as factors. The effects of groups [F (1, 36) = 67.18, p < 0.0001], training days [F (21, 756) = 7.411, p < 0.0001] and their interaction [F (21, 756) = 8.030, p < 0.0001] were significant (Supplementary Fig. 1A). During the first punishment phase, some rats (n = 12) exhibited continuous compulsive METH intake despite increasing shock intensity from 0.18 to 0.36 mA over 8 days; these rats were termed always shock-resistant (ASR) or compulsive drug takers (Fig. 1B). The observations of persistent resistance to punishment are comparable to the results of Giuliano et al. (2018) who had reported that compulsive alcohol seeking emerged in a subpopulation of rats in the presence of footshock punishment and persisted for almost a year [46]. In contrast to the punishment-resistant animals, other rats (n = 14) decreased lever pressing during the punishment phase and were named shock-sensitive (SS, n = 14) or non-compulsive (Fig. 1B). Rats were classified as shock-sensitive if they reduced their intake by more than 60% [33].

We used these two different METH phenotypes (ASR and SS) to conduct further statistical analysis using two-way ANOVA for the first 22 days of METH SA. The analysis revealed significant effects of training days [F (21, 504) = 17.67, p < 0.0001], but no significant effect of groups (ASR, SS) [F (1, 24) = 0.3708, p < 0.5483] nor their interaction [F (21, 504) = 1.283, p < 0.1798] (Fig. 1B and Supplementary Fig. 1B). When we analyzed the behavior for ASR and SS during the first footshock phase, we observed significant effects of groups [F (1, 244) = 32.53, p < 0.0001], footshock days [F (10, 240) = 28.23, p < 0.0001] and their interaction [F (10, 240) = 5.493, p < 0.0001] (Fig. 1B and Supplementary Fig. 1C).

Compulsive METH takers showed increased propensity to relapse

At the end of the first punishment phase, rats underwent METH-seeking tests under extinction conditions at withdrawal days (WD) 1 and WD15. Drug or food-seeking has been shown to increase gradually during periods of abstinence, a behavioral phenomenon termed “incubation of craving” [47–49]. These behaviors are measured by recording number of active lever presses in the presence of cues previously paired with METH infusions. Drug-seeking gradually increases during the period of abstinence and is clinically relevant because similar phenomena have been observed in human METH users [50]. Two-way ANOVA revealed significant effects for WD [F (1, 24) = 44.14, p < 0.0001], but only a significant trend for groups (CT, ASR, SS) [F (1, 24) = 3.790, p = 0.0634] and their interaction [F (1, 24) = 3.790, p = 0.0768]. Compulsive rats also displayed higher active lever pressing than non-compulsive rats on WD15 (Fig. 1C).

Some non-compulsive (SS) rats display compulsive METH use upon re-exposure to punishment after 15 days of forced abstinence

After the first abstinence period, rats were again allowed to self-administer METH. Both ASR and SS rats significantly increased their METH intake over the period of 12 days of METH SA (Fig. 1H). Rats were then re-exposed to a second contingent shock phase (shock intensity of 0.30 mA). As per above, rats that reduced their METH intake by more than 60% were classified as shock-sensitive [33]. Unexpectedly, we found that about 36% (n = 5) of the SS rats did not suppress their METH intake as they did during the first footshock phase (Fig. 1D); we labeled these animals ‘delayed shock-resistant’ (DSR) rats (Fig. 1H). The rest of SS rats continued to suppress their METH intake (Fig. 1H) as they did before (Fig. 1D) and we labeled those rats, ‘Always sensitive’ (AS), to separate them from the DSR and the SS rats. Importantly, the initially resistant rats (ASR) continued to self-administer METH in a compulsive fashion despite the footshocks (Fig. 1H).

The behavioral breakdown during the second punishment phase led us to re-analyze the data using saline and three METH self-administering phenotypes (ASR, DSR, and AS) as variables. We found that, during the first 22 days of SA, there were significant effects for groups (ASR, DSR and AS) [F (3, 34) = 23.97, p < 0.0001], training days [F (4.054, 137.8) = 21.64, p < 0.0001] and group by training days interaction [F (63, 714) = 4.767, P < 0.0001] (Fig. 1D).

Analysis of the first punishment phase documented that ASR rats continued to self-administer METH whereas DSR and AS rats decreased their METH intake in the presence of footshocks. ANOVA revealed significant effects for groups (ASR, DSR and AS) [F (2, 23) = 22.67, p < 0.0001], footshock days [F (3.562, 81.92) = 5.205, p = 0.0014] and their interaction [F (14, 161) = 2.408, p = 0.0044] (Fig. 1D). ASR rats showed significant differences in their METH SA in comparison to DSR and AS rats (Fig. 1D). In addition, total METH intake was significantly higher for ASR rats in comparison to DSR and AS rats during the last three days of footshocks (Fig. 1E).

METH seeking behaviors for ASR, DSR and AS rats were measured during forced abstinence. ANOVA revealed significant effects for withdrawal day [F (1, 34) = 42.79, p < 0.0001], groups (CT, ASR, DSR, and AS) [F (3, 34) = 5.532, p = 0.0033] and their interaction [F (3, 34) = 6.512, p = 0.0013]. ASR, DSR, and AS rats significantly increased their active level responding on WD15 in comparison to WD1 (Fig. 1F). ASR rats maintained significantly higher active lever responding in comparison to AS but not DSR rats (Fig. 1F). Regression analysis between METH intake during footshock phase and active lever responses on WD15 revealed a positive correlation (r = 0.6630, p < 0.0001) (Fig. 1G).

ANOVA for the second METH SA phase revealed significant effects for groups (CT, ASR, DSR and AS) [F (3, 34) = 40.58, p < 0.0001], SA days [F (11, 374) = 19.05, p < 0.0001] and SA days x groups interaction [F (33, 374) = 4.963, p < 0.0001] (Fig. 1H). The total METH intake was also significantly higher in ASR rats when compared to AS rats (Supplementary Fig. 1D). There were no significant differences in total METH intake between ASR vs DSR for that phase (Supplementary Fig. 1D).

ANOVA for the second punishment phase identified significant effects for groups [F (2, 23) = 19.30, p < 0.0001], punishment days [F (1.882, 43.28) = 22.32, p < 0.0001] and their interaction [F (6, 69) = 3.374, p = 0.0056]. Both ASR and DSR rats continue to self-administer METH despite footshock punishment and self-administered significantly higher METH infusion during the three days of footshocks in comparison to AS rats (Fig. 1H). ASR and DSR had significantly higher total METH intake than AS rats during three days of footshocks (Fig. 1I).

Persistently compulsive METH takers continued to show greater propensity to relapse

Relapse tests were conducted for a second time after forced abstinence. ANOVA revealed significant effects of withdrawal day [F (1, 34) = 22.58, p < 0.0001], groups [F (3, 34) = 3.183, p = 0.0362] and their interaction [F (3, 34) = 3.411, p = 0.0283]. Post hoc test showed that ASR, DSR and AS rats significantly increased their active level responding on WD15 compared to WD1 (Fig. 1J). ASR rats had significant higher active lever responding on WD15 in comparison to CT and AS animals but not DSR rats (p = 0.0757) (Fig. 1J). There were positive correlations between active lever responses on WD15 of the second set of relapse tests against total METH intake during the second footshock phase (Fig. 1K, r = 0.4960, p < 0.0015) and total METH intake for all rats throughout the experiment (Supplementary Fig. 1E,r =  0.5170, p < 0.0009).

Based on these data, we reasoned that these behavioral differences might be associated with molecular differences in brain regions that subsume habit forming and/or acquisition of sequential behaviors. We thus performed RNA sequencing to identify potential transcriptional changes in the dorsal striatum that might be specifically associated with the different behaviors observed in the three groups of METH rats.

RNA sequencing identifies specific differentially expressed genes in the dorsal striatum of always-resistant, delayed-resistant, and always-sensitive rats

To identify potential gene networks involved in compulsive and suppressed behaviors in response to punishment, we used RNA sequencing to measure global transcriptional changes in the dorsal striatum (dSTR) of rats euthanized 24 h after the second WD15 relapse test. Results of RNA sequencing are illustrated in Fig. 2. Using DESeq2, we performed comparisons of gene expression between eight different pair-wise comparison (ASR vs CT, DSR vs CT, AS vs CT, ASR vs AS, DSR vs AS, ASR vs DSR, AS vs ASR, and AS vs DSR); the results are shown as volcano plots in Fig. 2. Analysis of raw sequencing data using Log2 fold changes and log10 p values revealed the number of genes with higher or lower expression in ASR vs CT (Fig. 2A), DSR vs CT (Fig. 2B), AS vs CT (Fig. 2C), ASR vs AS (Fig. 2D), DSR vs AS (Fig. 2E), ASR vs DSR (Fig. 2F), AS vs ASR (Fig. 2G), and for AS vs DSR (2H) comparisons.Fig. 2 RNA sequencing revealed large-scale changes in gene expression in the dorsal striatum.

Analysis of raw sequencing data using Log2 fold changes and log10 p values revealed many differentially expressed genes in pairwise comparisons shown as volcano plots: (A) ASR vs CT, (B) DSR vs CT, (C) AS vs CT, (D) ASR vs AS, (E) DSR vs AS, (F) ASR vs DSR, (G) AS vs ASR and (H) AS vs DSR. I Hierarchical clustering of 1194 differentially expressed genes across six pair-wise comparisons. The red color represents over-expressed genes, the blue color represents genes with reduced expression, while the yellow color represents genes with no changes in expression. J Functional gene clusters and (K) KEGG analysis show pathway distribution of differentially expressed genes according to DAVID.

We also used a more restrictive cut-off of equal or greater than 1.5-fold (p = 0.05) and used Qiagen Ingenuity Pathway software to identify gene networks that might have been affected in the different groups of METH rats. The number of differentially impacted genes is provided for ASR vs CT, DSR vs CT, AS vs CT, ASR vs AS, DSR vs AS, and ASR vs DSR in the Venn diagrams (Supplementary Fig. 2A, B) and the changes in the expression of these 1194 genes are illustrated in the hierarchical clustering heat-map shown in Fig. 2I.

The Database for Annotation, Visualization and Integrated Discovery (DAVID) was also used to generate functional annotation and clustering for these 1194 differentially expressed genes (DEGs). Functional gene clusters and KEGG pathways are illustrated in Figs. 2J, K. IPA network for the ASR vs CT, DSR vs CT and AS vs CT comparisons were shown in Fig. 3A, B, and C.Fig. 3 Ingenuity pathway analysis (IPA) identifies several networks of DEGs in compulsive and non-compulsive METH takers in comparison to the control groups.

A Pathways and genes significantly enriched in the ASR vs CT comparison. B Pathways and genes significantly enriched in the DSR vs CT comparison. C Pathways and genes significantly enriched in the AS vs CT comparison. The red color indicates upregulated genes, the green color represents downregulated genes, and the blue color represent interacting gene partners.

Specific striatal genes are differentially expressed in rats prone to relapse

One of the important problems associated with the clinical course of MUD is frequent relapses. Relapses have been modeled in animal models using the incubation of drug craving paradigm [43, 47, 48, 51, 52]. In our study, ASR rats showed greater incubation of METH craving that the DSR and AS rats. We used pairwise comparisons to identify genes that were uniquely expressed in ASR rats in comparison to other groups (Fig. 4A). Among these genes are Mybpc1, Dipk2b, Rab3, and Tnfsf8. Figure 4B illustrates IPA analysis for genes uniquely found in ASR vs DSR. Supplementary Fig. 3B, C also show IPA analyses for the 180 unique genes in the ASR vs AS comparison. The DEGs uniquely impacted in the ASR groups participate in learning, cognition, addictive behavior, and METH dependence. These results are consistent with behaviors like cognitive impairments reported in patients with diagnosis of MUD [53].Fig. 4 IPA analysis of unique DEGs in the dorsal striatum of ASR, DSR, and AS rats.

The Venn diagram in (A) illustrates common and unique upregulated gene between 3 pairwise comparisons ASR vs (CT, DSR and AS). B Networks significantly enriched among 82 unique genes in the ASR vs DSR comparison. C The Venn diagram illustrates the distribution of 50 genes with the highest fold in the ASR compared to DSR or AS rats. D IPA analysis shows that 34 shared between ASR vs AS and DSR vs AS comparisons are involved in processes that mediate memory, learning, cognition, and METH dependence. Cognitive impairments are common in patients with MUD. E The Venn diagram illustrates the number of genes that are commonly and uniquely located in 3 pairwise comparisons of AS vs (CT, ASR and DSR). F IPA shows networks involving 101 genes that are common in AS vs CT, AS vs ASR, and AS vs DSR comparisons (71 genes) and in AS vs ASR and AS vs DSR comparisons (31 genes). These genes are involved in amphetamine delusional disorder, cognitive impairments, and addictive behaviors. The red color indicates upregulated genes, and the blue color represents interacting genes.

Figure 4C shows 34 genes with higher expression that were common in the ASR and DSR groups in comparison to the AS group. Figure 4D and Supplementary Fig. 4 illustrate the involvement of several of these genes in learning and memory formation, cognitive processes known to be impaired in MUD patients [53].

Specific striatal genes are differentially expressed in always sensitive (AS) rats

We also sought to identify genes that were impacted in the always sensitive (AS) rats. We used the following comparisons: AS vs CT, AS vs ASR, and AS vs DSR, and identified 71 common that had higher expression in the AS group in comparison to the other group (Fig. 4E). These genes included Kcnk16, Cfap43, Cfap44, DnaH1, DnaH7, and DnaI2, among others. Thirty genes with higher expression in the AS in comparison to ASR and DSR groups are also shown in Fig. 4E). We reasoned that these genes might be involved in inhibiting drug taking behaviors in the presence of adverse consequences. The IPA analysis showed the involvement of some of these genes in learning, cognition, depression, delusional disorder, and addictive behavior (Fig. 4F).

PCR validation of some genes identified by RNA sequencing analysis

We opted to use quantitative PCR to validate the expression of some of the genes of interest identified in the RNA sequencing data (Fig. 5). ASR rats displayed significant higher levels of Rab37 [F (3, 19) = 14.68, p < 0.0001] (Fig. 5A), Dipk2b [F (3, 19) = 10.45, p = 0.0003] (Fig. 5B), and Mybpc1 [F (3, 19) = 5.733, p = 0.0057] (Fig. 5C) than CT, DSR, and AS. In contrast, Tnfsf8 expression was decreased [F (3, 19) = 9.230, p = 0.0006] (Fig. 5D) in ASR in comparison to CT, DSR, and AS rats. Another member of the Dipk2b family, Dipk2a showed increased [F (3, 19) = 5.610, p = 0.0063] (Fig. 5E) expression in DSR compared to CT and AS. In addition, mRNA levels for Mybpc3 [F (3, 19) = 5.346, p = 0.0077] (Fig. 5F), but not of Mybpc2 (data not shown), were significantly increased in ASR and DSR rats. Because a previous paper had reported that Mybpc1 was expressed in VIP-expressing interneurons [54], we measured Vip mRNA levels and found significant increases [F (3, 19) = 8.871, p = 0.0007] only in DSR rats (Fig. 5G).Fig. 5 PCR validation of genes identified by RNA sequencing analysis as differentially expressed in various pairwise comparisons between the four groups of rats.

Expression of (A) Rab37, (B) Dipk2b, (C) Mybpc1, (D) Tnfsf8, (E) Dipk2a, (F) Mybpc3, (G) Vip, (H) Foxn3, (I) Cartpt, (J) Avp, (K) Hcrtr1, (L) Hcrtr2, (M) Bdnf, (N) Ngf, (O) TrkA, (P) TrkB, (Q) TrkC, (R) p75ntr, (S) Fgf1, (T) Fgf2, (U) Tgfa, (V) Tgfb1, (W) Vegfa, (X) Tnf, (Y) Kcnk16, (Z) Cfap43, (AA) Cfap44, (AB) DnaH1, (AC) DnaH7, and (AD) DnaI2 in CT, saline; ASR, always shock-resistant; DSR, delayed shock-resistant; and AS, always sensitive rats. Key to statistics: *p < 0.05, **p < 0.01, ***p < 0.001, comparisons between METH groups (ASR, DSR, AS) and controls (CT); #p < 0.05, ##p < 0.01, ###p < 0.001, comparison between ASR and AS; ! p < 0.05, !!p < 0.01, !!!p < 0.001, comparison between ASR and DSR; $p < 0.05, $$p < 0.01, $$$p < 0.001, comparison between DSR and AS.

Regression analysis between active lever presses in the second WD15 relapse test and PCR-validated genes revealed positive correlations for Dipk2b (r = 0.4514, p = 0.0306) and Rab37 (r = 0.5614, p = 0.0053) and a negative correlation for Tnfsf8 (r = −0.4368, p = 0.0371) (Supplementary Fig. 5A, B, and C, respectively).

We also used PCR to validate Foxn3, Cartpt, Avp, and Hcrtr whose mRNA levels were increased in both compulsive ASR and DSR rats in comparison CT and AS animals (Fig. 3, A, B). These are shown in Fig. 5H–L.

Striatal Bdnf mRNA levels showed decreased [F (3, 19) = 4.397, p = 0.0165] mRNA expression in AS rats in comparison to the other groups, consistent with the RNA-Seq data (Fig. 5M). Although they were not identified in the RNA sequencing analysis, we opted to measure the expression of some other trophic factors, because members of these trophic factors have been implicated in various SUDs [37, 42, 55, 56]. Changes in their expression might also be related to relapse [57]. The results for these including for Fgf1 and Fgf2 are shown in Fig. 5N–X (see below for statistical analyses). Interestingly, AS rats also exhibited significant increases in Kcnk16, Cfap43, Cfap44, DnaH1, DnaH7, and DnaI2 mRNA levels in comparison to the other groups (Fig. 5Y–AD), suggesting that their increased expression might be related to suppression of METH taking behaviors in the presence of punishment and/or decreased propensity to relapse.

Trophic factor-related genes are differentially expressed in compulsive METH users

As mentioned above, there were significant decreases in Bdnf (Fig. 5M) expression in the AS METH groups. In order to provide a more panoramic view of potential changes in these families of trophic factors in the METH groups, we used quantitative PCR to measure the expression of some members of the FGF family of genes that have been reported to be impacted in other models of addiction [42, 58]. Fgf1 mRNA levels were increased [F (3, 19) = 4.429, p = 0.0160] in both ASR and DSR phenotypes (Fig. 5S). Fgf2 was increased significantly [F (3, 19) = 5.016, p = 0.0100] in DSR (Fig. 5T). Elevated levels of Tgfb1 [F (3, 19) = 4.936, p = 0.0106] were seen in DSR and ASR in comparison to AS (Fig. 5V). Moreover, DSR rats showed significantly higher [F (3, 19) = 3.896, p = 0.0252] Vegfa mRNA levels in comparison to AS and control rats (Fig. 5W). Tnf mRNA levels were significantly increased [F (3, 19) = 6.547, p = 0.0032] in ASR, DSR, and AS in comparison to CT rats (Fig. 5X). In contrast, Tgfa (Fig. 5U), Egf, Igf1, Tgfb2, Tgfb3, and Fgf9 mRNAs were not impacted by METH SA and shocks (data not shown).

The expression of trophic factor genes is regulated by HDAC2 recruitment at their promoters

IPA analysis had revealed a potential involvement of histone deacetylases in the regulation of some genes relevant to compulsive drug taking and/or abstinence (see Fig. 3). We had also shown previously that HDAC2 participated in the regulation of METH-induced changes in gene expression in mice [59, 60]. We thus decided to further test the role of HDAC2 in the regulation of some genes of interest by performing chromatin immunoprecipitation (ChIP) followed by qPCR. There were significant decreases [F (3, 24) = 3.040, p = 0.0484] in HDAC2 binding at the VIP promoter regions in ASR and DSR (Fig. 6A); these results are partially consistent with the mRNA data that showed increased VIP expression in DSR rats. There were also significant decreases [F (3, 20) = 3.167, p = 0.0469] in HDAC2 binding at the AVP promoter sequence in the ASR and DSR groups (Fig. 6B) in a manner consistent with the mRNA data (see Fig. 5J). HDAC2 binding at HCRTR2 was also increased [F (3, 17) = 5.026, p = 0.0113] in ASR and DSR (Fig. 6C).Fig. 6 Chromatin immunoprecipitation (ChIP) using a specific HDAC2 antibody identifies several HDAC2-regulated genes in the ASR, DSR, and AS rats.

Fold enrichment of: (A) VIP, (B) AVP, (C), HCRTR2, (D) BDNF, (E) TrkB, (F) FGF1, (G) FGF2, (H) VEGFa. CT, saline; ASR, always shock-resistant; DSR, delayed shock-resistant; and AS, always sensitive rats. Keys to statistics are as described in Fig. 5.

Interestingly and importantly, we found significantly increased [F (3, 21) = 6.889, p = 0.0021] HDAC2 enrichment at the BDNF promoter sequence in AS animals (Fig. 6D); these observations are consistent with the decreased Bdnf mRNA levels in the same rats (Fig. 5M). There was also decreased [F (3, 19) = 40142, p = 0.0204] HDAC2 enrichment at the TrkB promoter sequence for ASR and DSR (Fig. 5E), in a manner consistent with the increased mRNA levels in those groups. Other genes for trophic factors showed some interesting changes at their promoters, with FGF1 showing decreased [F (3, 23) = 3.484, p = 0.0322] HDAC2 binding for ASR and DSR rats (Fig. 6F) consistent with FGF1 mRNA levels (Fig. 5S), FGF2 showing significantly increased [F (3, 24) = 7.403, p = 0.0011] HDAC2 binding in AS rats (Fig. 6G), and VEGFa showing decreased [F (3, 20) = 6.691, p = 0.0026] HDAC2 recruitment in the ASR and DSR (Fig. 6H) groups. The ChIP-PCR data are therefore consistent with the mRNA data (Fig. 5W).

Discussion

The primary aim of the present study was to assess the longitudinal course of compulsivity and/or abstinence in METH SA rats in response to footshock punishment and to measure the rats’ propensity to relapse during forced abstinence. A secondary aim was to measure global gene expression in the dorsal striatum of the METH SA rats in order to identify specific networks of genes that might be associated with the different behavioral phenotypes. The study confirmed the existence of rats that displayed compulsive METH taking throughout the behavioral experiment despite punishment. These differential responses to METH SA and punishment are consistent with the accumulated evidence that rats show individual differences in their responses to psychostimulants like cocaine and the amphetamines [61–65]. The individual responses to drug self-administration become more prominent in the presence of contingent footshocks [62, 66]. In addition, we found, for the first time, that thirty-six (36%) percent of rats that were initially punishment-sensitive became compulsive METH takers after a second round of METH SA. As previously reported [31, 33], persistently compulsive rats showed significantly higher incubation of METH craving than non-compulsive rats, suggesting that different molecular mechanisms were in play in the brains of these rats. Our RNA sequencing data which identified several genes whose mRNA levels were impacted differentially in the ASR, DSR, and AS rats support this notion, with Dipk2b and Rab37 mRNA levels showing significant positive correlation with incubation of METH craving.

Development of early and late compulsive behaviors

Rats that showed resistance to footshocks during the first phase of the behavioral experiment remained punishment-resistant even after a period of 2 weeks of forced abstinence. Interestingly, some punishment-sensitive rats became resistant to the footshocks during the second phase of the experiment, supporting the notion that some individuals may be more sensitive to the addictive effects of amphetamine-type substances [67, 68]. The development of delayed resistance to shocks is like the resurgence of behaviors that had been suppressed by punishment or other means [69–74]. Our results are also consistent with the data of Kashani et al. (1987) who had reported that several of their patients had developed SUDs earlier than other patients [75]. Moreover, our behavioral data are compatible with the documentation of two multi-dimensional subtypes of patients who suffer from SUDs [76] including alcohol [77], cocaine [78], and METH [79] use disorders. Altogether, these findings support clinical observations that subpopulations of patients who misuse psychostimulants do exhibit variable clinical courses which might be secondary to different molecular mechanisms. This reasoning suggests the possibility of different approaches to mitigate relapses [72, 73, 80].

Persistent compulsive METH-taking behavior and proneness to relapse

Rats that showed persistent resistance to punishment throughout the experiment also exhibited greater incubation of METH craving as previously reported [31, 37, 59]. Unexpectedly, rats that developed delayed compulsive METH-taking behaviors during the second set of footshocks failed to show incubation of METH craving as seen in the persistently compulsive rats (see Fig. 1J). These differences in behaviors are supported by the RNA sequencing data that identified genes whose mRNA levels were altered only in the persistently compulsive rats (see Fig. 5A–D). These DEGs include Dipk2b, Rab37, and Mybpc1 that showed increased mRNA levels, changes that are correlated to the magnitude of lever pressing for METH during the second set of relapse tests (Supplementary Fig. 5A, B, and D). Dipk2b has been reported to alter neuronal cellular secretory pathways that are involved in autism [81, 82] while RAB37 is known to participate in membrane trafficking and in the regulation of TIMP1 exocytosis [83]. Mybpc1 is thought to contribute to changes in synaptic plasticity [54], which is thought to be an important mediator of the impact of rewarding substances on the brain [7–9, 11]. More experiments, in which the expression of these genes is manipulated, are needed to identify the specific roles that they might play in the development of compulsive METH taking behaviors and incubation of METH craving.

Delayed METH taking and molecular mechanisms

We identified significant increases in Dipk2a, Avp, Fgf2, Tgfb1 and Vegfa mRNA levels in the dorsal striatum of rats that show delayed resistance to footshocks after METH SA behaviors were suppressed by punishment. The changes in Avp mRNA levels are of interest because AVP is known to regulate stress-related behaviors [84, 85] and has been implicated in the development of substance use disorders [86]. The present observations are consistent with those of other studies that had documented increased Avp mRNA expression in the nucleus accumbens after exposure to METH [45, 87] and in the amygdala after heroin SA and footshocks [88].

There is some documentation of the participation of trophic factors in the manifestation of drug-taking behaviors. For example, the role for Fgf2 in SUDs has been well documented [58]. Interestingly, oxycodone self-administering rats showed higher Fgf2 levels in the dorsal striatum after a month of withdrawal [42]. Of related interest, endogenous FGF2 expression is necessary for the development of sensitization to amphetamine [89]. In addition, the expression of another trophic factor, Vegfa, identified in our present study was also increased in the nucleus accumbens of rats after 4 weeks of cocaine administration [90]. Moreover, plasma TGF-beta1 levels were reported to be increased in patients with alcohol use disorders [91]. Altogether, these observations support the idea of the involvement of trophic factors in several models of SUDs.

Non-compulsive behavior and potential therapeutic approaches

Identifying genes and molecular mechanisms involved in METH addiction is necessary because there is presently no FDA-approved medication for MUD. In the present study, we observed increases in the mRNA levels of potassium channel Kcnk16, cilia and flagella associated protein 43 (Cfap43) [92], and dynein (DnaH1, DnaH7, and DnaI2) [93–96] in the subpopulation of METH SA rats that always reduced their METH intake in the presence of punishment. The increased expression of Kcnk16 is consistent with the results of our previous experiments in which we also identified increased mRNA expression of potassium channels in shock-sensitive rats [28]. These changes are consistent with findings that potassium channel activators can reduce the intake of rewarding substances (reviewed in 95). Although the potential roles of dynein have not been investigated extensively in models of SUDs, it was recently reported that rats which were chronically injected with heroin showed decreased cortical dynein protein expression [96]. It remains to be determined to what extent manipulations of these genes might influence METH self-administration in the presence of adverse consequences.

HDAC2 recruitment regulated the expression of genes involved in compulsive behavior

Epigenetic mechanisms that include posttranslational modifications of histone residues participate in the regulation of the expression of plasticity genes that might be responsible for SUD development [26, 97–100]. Histone deacetylases, including HDAC2, that remove acetyl groups from histone residues participate in the regulation of genes that are involved in memory formation and synaptic plasticity [101]. Because HDACs participate in the behavioral manifestations observed in SUD models in animals [26, 102, 103] and in the regulation of the expression of METH-induced immediate early genes in the nucleus accumbens [59], we had reasoned that HDAC2 might regulate the expression of some of the genes identified in the RNA sequencing analysis.

To test the idea, we ran ChIP-PCR to analyze HDAC2 recruitment at some of the mRNAs that we had validated by qPCR. We found that the expression of several trophic factor-related genes including Bdnf, TrkB, Fgf1, Fgf2 and Vegfa showed evidence of regulation by HDAC2 binding at their promoters. Specifically, we found significant increases in HDAC2 binding at the BDNF promoter (see Fig. 6D) and marked decreases in Bdnf mRNA levels in the dorsal striatum of AS rats that remained sensitive to the effects of punishment throughout the experiment (see Fig. 5M). The present observations are consistent with those of Guan et al. (2009) who had reported that Bdnf expression was regulated by HDAC2, with loss of HDAC2 causing increased Bdnf expression in the mouse brain [104]. Moreover, blocking endogenous BDNF expression in nucleus accumbens reduces cocaine SA and relapse [105]. Partial knockout of BDNF also attenuated cocaine seeking behavior in rats [106]. Altogether, these observations support targeting BDNF expression as a potential treatment strategy against MUD.

In addition, we found decreased HDAC2 recruitment at the TrkB promoter which was associated in increased TrkB mRNA expression in both compulsive (ASR and DSR) groups. Similar observations were made for Fgf1 that shows increased mRNA expression in both compulsive (ASR and DSR) rats. Taken together, these data support the involvement of HDAC2 which is known to regulate synaptic plasticity as well as learning and memory processes [101, 104, 107] in regulating the expression of genes that appear to be important in the maintenance of compulsive METH taking in the presence of adverse consequences.

Conclusion

Our study reports, for the first time shows, that some animals that had initially suppressed their METH intake during a first round of footshocks became resistant to the effects of a second round of punishment. Rats that were resistant to footshocks throughout the experiment showed more incubation of METH craving than other rats. RNA sequencing analysis identified increased expression of some genes including Rab37 and Dipk2b in persistent resistant rats, with their expression showing positive correlation to lever pressing during relapse tests. We also identified increased expression of Kcnk16, Cfap43, DnaH1, DnaH7, and DnaI2 in always sensitive animals. ChIP-PCR identified HDAC2 as a regulator of Bdnf, TrkB, and Fgf1 mRNA levels. Thus, our data are of significant interest because they provide a model that is relevant to the clinical transition from recreational drug taking to compulsive drug taking and addiction that occur with different time courses in human METH users. The RNA sequencing data also identified potential targets for pharmacological interventions against MUD.

Supplementary information

Supplementary file

Supplementary information

The online version contains supplementary material available at 10.1038/s41380-024-02455-2.

Acknowledgements

This work is supported by the Department of Health and Human Services/National Institutes of Health/National Institute on Drug Abuse/Intramural Research Program, Baltimore, MD, USA.

Author contributions

Conceptualization, J.L.C., A.P.D. and S.J.; Investigation, A.P.D., M.T.M., B.L. and S.J.; Methodology, A.P.D., M.T.M., B.L. and S.J. Validation, M.T.M., B.L. and A.P.D.; Visualization, A.P.D. and M.T.M.; Formal analysis, A.P.D. and J.L.C.; Project administration, J.L.C. and A.P.D.; Supervision, J.L.C., Writing-original draft, A.P.D.; Writing-review and editing, A.P.D., M.T.M., B.L., S.J. and J.L.C. All authors have read and agreed to the published version of the manuscript.

Funding

This project was supported by the Intramural Research Program of the National Institute on Drug Abuse (NIDA), NIH, and DHHS [grant # DA000552 (2021)]. Open access funding provided by the National Institutes of Health.

Data availability

The RNA sequencing data have been deposited at the NCBI GEO under the accession # GSE220896. All other data generated in this study, including PCR, CHIP-PCR, Excel spreadsheets, and GraphPad files, are available upon reasonable request to the corresponding author via email.

Competing interests

The authors declare no competing interests.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
==== Refs
References

1. United Nations Office on Drugs and Crime (UNODC). “Drug Market Trends: Cocaine, Amphetamine-type stimulants. United Nations Publication” Vienna, Austria, 2021 E.21. XI. 8, 2021. https://www.unodc.org/unodc/en/data-and-analysis/wdr-2021_booklet-4.html.
2. American Psychiatric Association. “Diagnostic and statistical manual of mental disorders, (DSM-5-TR)”, 2013. https://www.psychiatry.org/psychiatrists/practice/dsm.
3. Han B Compton WM Jones CM Einstein EB Volkow ND Methamphetamine use, methamphetamine use disorder, and associated overdose deaths among us adults JAMA Psychiatry 2021 78 1329 42 10.1001/jamapsychiatry.2021.2588 34550301
Han B, Compton WM, Jones CM, Einstein EB, Volkow ND. Methamphetamine use, methamphetamine use disorder, and associated overdose deaths among us adults. JAMA Psychiatry. 2021;78:1329–42.34550301
4. Courtney KE Ray LA Methamphetamine: an update on epidemiology, pharmacology, clinical phenomenology, and treatment literature Drug Alcohol Depend 2014 143 11 21 10.1016/j.drugalcdep.2014.08.003 25176528
Courtney KE, Ray LA. Methamphetamine: an update on epidemiology, pharmacology, clinical phenomenology, and treatment literature. Drug Alcohol Depend. 2014;143:11–21.25176528
5. Anton RF Obsessive-compulsive aspects of craving: development of the obsessive-compulsive drinking scale Addiction 2000 95 S211 217 10.1046/j.1360-0443.95.8s2.9.x 11002915
Anton RF. Obsessive-compulsive aspects of craving: development of the obsessive-compulsive drinking scale. Addiction. 2000;95:S211–217.11002915
6. Figee M Pattij T Willuhn I Luigjes J van den Brink W Goudriaan A Compulsivity in obsessive-compulsive disorder and addictions Eur Neuropsychopharmacol 2016 26 856 68 10.1016/j.euroneuro.2015.12.003 26774279
Figee M, Pattij T, Willuhn I, Luigjes J, van den Brink W, Goudriaan A, et al. Compulsivity in obsessive-compulsive disorder and addictions. Eur Neuropsychopharmacol. 2016;26:856–68.26774279
7. Lüscher C Robbins TW Everitt BJ The transition to compulsion in addiction Nat Rev Neurosci 2020 21 247 63 10.1038/s41583-020-0289-z 32231315
Lüscher C, Robbins TW, Everitt BJ. The transition to compulsion in addiction. Nat Rev Neurosci. 2020;21:247–63.32231315
8. Volkow ND Wang GJ Fowler JS Tomasi D Addiction circuitry in the human brain Annu Rev Pharmacol Toxicol 2012 52 321 36 10.1146/annurev-pharmtox-010611-134625 21961707
Volkow ND, Wang GJ, Fowler JS, Tomasi D. Addiction circuitry in the human brain. Annu Rev Pharmacol Toxicol. 2012;52:321–36.21961707
9. Everitt BJ Neural and psychological mechanisms underlying compulsive drug seeking habits and drug memories–indications for novel treatments of addiction Eur J Neurosci 2014 40 2163 82 10.1111/ejn.12644 24935353
Everitt BJ. Neural and psychological mechanisms underlying compulsive drug seeking habits and drug memories–indications for novel treatments of addiction. Eur J Neurosci. 2014;40:2163–82.24935353
10. Cadet JL Brannock C Jayanthi S Krasnova IN Transcriptional and epigenetic substrates of methamphetamine addiction and withdrawal: evidence from a long-access self-administration model in the rat Mol Neurobiol 2015 51 696 717 10.1007/s12035-014-8776-8 24939695
Cadet JL, Brannock C, Jayanthi S, Krasnova IN. Transcriptional and epigenetic substrates of methamphetamine addiction and withdrawal: evidence from a long-access self-administration model in the rat. Mol Neurobiol. 2015;51:696–717.24939695
11. Cadet JL Epigenetics of stress, addiction, and resilience: therapeutic implications Mol Neurobiol 2016 53 545 60 10.1007/s12035-014-9040-y 25502297
Cadet JL. Epigenetics of stress, addiction, and resilience: therapeutic implications. Mol Neurobiol. 2016;53:545–60.25502297
12. Koob GF Volkow ND Neurobiology of addiction: a neurocircuitry analysis Lancet Psychiatry 2016 3 760 73 10.1016/S2215-0366(16)00104-8 27475769
Koob GF, Volkow ND. Neurobiology of addiction: a neurocircuitry analysis. Lancet Psychiatry. 2016;3:760–73.27475769
13. Krasnova IN Gerra MC Walther D Jayanthi S Ladenheim B McCoy MT Compulsive methamphetamine taking in the presence of punishment is associated with increased oxytocin expression in the nucleus accumbens of rats Sci Rep 2017 7 8331 10.1038/s41598-017-08898-8 28827541
Krasnova IN, Gerra MC, Walther D, Jayanthi S, Ladenheim B, McCoy MT, et al. Compulsive methamphetamine taking in the presence of punishment is associated with increased oxytocin expression in the nucleus accumbens of rats. Sci Rep. 2017;7:8331.28827541
14. Pascoli V Hiver A Van Zessen R Loureiro M Achargui R Harada M Stochastic synaptic plasticity underlying compulsion in a model of addiction Nature 2018 564 366 71 10.1038/s41586-018-0789-4 30568192
Pascoli V, Hiver A, Van Zessen R, Loureiro M, Achargui R, Harada M, et al. Stochastic synaptic plasticity underlying compulsion in a model of addiction. Nature. 2018;564:366–71.30568192
15. McDonald AJ Alonso-Lozares I Rauh V van Mourik Y Schetters D De Vries TJ Alcohol seeking under risk of punishment is associated with activation of cortical and subcortical brain regions Front Behav Neurosci 2021 15 739681 10.3389/fnbeh.2021.739681 34744653
McDonald AJ, Alonso-Lozares I, Rauh V, van Mourik Y, Schetters D, De Vries TJ, et al. Alcohol seeking under risk of punishment is associated with activation of cortical and subcortical brain regions. Front Behav Neurosci. 2021;15:739681.34744653
16. Browne CJ Futamura R Minier-Toribio A Hicks EM Ramakrishnan A Martínez-Rivera FJ Transcriptional signatures of heroin intake and relapse throughout the brain reward circuitry in male mice Sci Adv 2023 9 eadg8558 10.1126/sciadv.adg8558 37294757
Browne CJ, Futamura R, Minier-Toribio A, Hicks EM, Ramakrishnan A, Martínez-Rivera FJ, et al. Transcriptional signatures of heroin intake and relapse throughout the brain reward circuitry in male mice. Sci Adv. 2023;9:eadg8558.37294757
17. Knowlton BJ Patterson TK Habit formation and the striatum. Curr Top Behav Neurosci 2018 37 275 95 10.1007/7854_2016_451 27677776
Knowlton BJ, Patterson TK.Habit formation and the striatum.Curr Top Behav Neurosci. 2018;37:275–95.27677776
18. Williamson LL Cheng RK Etchegaray M Meck WH Speed” warps time: methamphetamine’s interactive roles in drug abuse, habit formation, and the biological clocks of circadian and interval timing Curr Drug Abuse Rev 2008 1 203 12 10.2174/1874473710801020203 19630719
Williamson LL, Cheng RK, Etchegaray M, Meck WH. Speed” warps time: methamphetamine’s interactive roles in drug abuse, habit formation, and the biological clocks of circadian and interval timing. Curr Drug Abuse Rev. 2008;1:203–12.19630719
19. Caprioli D Venniro M Zhang M Bossert JM Warren BL Hope BT Role of dorsomedial striatum neuronal ensembles in incubation of methamphetamine craving after voluntary abstinence J Neurosci 2017 37 1014 27 10.1523/JNEUROSCI.3091-16.2016 28123032
Caprioli D, Venniro M, Zhang M, Bossert JM, Warren BL, Hope BT, et al. Role of dorsomedial striatum neuronal ensembles in incubation of methamphetamine craving after voluntary abstinence. J Neurosci. 2017;37:1014–27.28123032
20. Furlong TM Corbit LH Brown RA Balleine BW Methamphetamine promotes habitual action and alters the density of striatal glutamate receptor and vesicular proteins in dorsal striatum Addict Biol 2018 23 857 67 10.1111/adb.12534 28707389
Furlong TM, Corbit LH, Brown RA, Balleine BW. Methamphetamine promotes habitual action and alters the density of striatal glutamate receptor and vesicular proteins in dorsal striatum. Addict Biol. 2018;23:857–67.28707389
21. Lipton DM Gonzales BJ Citri A Dorsal striatal circuits for habits, compulsions, and addictions Front Syst Neurosci 2019 13 28 10.3389/fnsys.2019.00028 31379523
Lipton DM, Gonzales BJ, Citri A. Dorsal striatal circuits for habits, compulsions, and addictions. Front Syst Neurosci. 2019;13:28.31379523
22. Vaillancourt K Yang J Chen GG Yerko V Théroux JF Aouabed Z Cocaine-related DNA methylation in caudate neurons alters 3D chromatin structure of the IRXA gene cluster Mol Psychiatry 2021 26 3134 51 10.1038/s41380-020-00909-x 33046833
Vaillancourt K, Yang J, Chen GG, Yerko V, Théroux JF, Aouabed Z, et al. Cocaine-related DNA methylation in caudate neurons alters 3D chromatin structure of the IRXA gene cluster. Mol Psychiatry. 2021;26:3134–51.33046833
23. Pascoli V Hiver A Li Y Harada M Esmaeili V Lüscher C Cell-type specific synaptic plasticity in dorsal striatum is associated with punishment-resistance compulsive-like cocaine self-administration in mice Neuropsychopharmacology 2023 48 448 58 10.1038/s41386-022-01429-8 36071131
Pascoli V, Hiver A, Li Y, Harada M, Esmaeili V, Lüscher C. Cell-type specific synaptic plasticity in dorsal striatum is associated with punishment-resistance compulsive-like cocaine self-administration in mice. Neuropsychopharmacology. 2023;48:448–58.36071131
24. McNally GP Jean-Richard-Dit-Bressel P Millan EZ Lawrence AJ Pathways to the persistence of drug use despite its adverse consequences Mol Psychiatry 2023 28 2228 37 10.1038/s41380-023-02040-z 36997610
McNally GP, Jean-Richard-Dit-Bressel P, Millan EZ, Lawrence AJ. Pathways to the persistence of drug use despite its adverse consequences. Mol Psychiatry. 2023;28:2228–37.36997610
25. Cadet JL Patel R Jayanthi S Compulsive methamphetamine taking and abstinence in the presence of adverse consequences: Epigenetic and transcriptional consequences in the rat brain Pharmacol Biochem Behav 2019 179 98 108 10.1016/j.pbb.2019.02.009 30797763
Cadet JL, Patel R, Jayanthi S. Compulsive methamphetamine taking and abstinence in the presence of adverse consequences: Epigenetic and transcriptional consequences in the rat brain. Pharmacol Biochem Behav. 2019;179:98–108.30797763
26. Cadet JL Jayanthi S Epigenetic landscape of methamphetamine use disorder Curr Neuropharmacol 2021 19 2060 6 10.2174/1570159X19666210524111915 34030618
Cadet JL, Jayanthi S. Epigenetic landscape of methamphetamine use disorder. Curr Neuropharmacol. 2021;19:2060–6.34030618
27. Jayanthi S Daiwile AP Cadet JL Neurotoxicity of methamphetamine: Main effects and mechanisms Exp Neurol 2021 344 113795 10.1016/j.expneurol.2021.113795 34186102
Jayanthi S, Daiwile AP, Cadet JL. Neurotoxicity of methamphetamine: Main effects and mechanisms. Exp Neurol. 2021;344:113795.34186102
28. Cadet JL Brannock C Krasnova IN Jayanthi S Ladenheim B McCoy MT Genome-wide DNA hydroxymethylation identifies potassium channels in the nucleus accumbens as discriminators of methamphetamine addiction and abstinence Mol Psychiatry 2017 22 1196 204 10.1038/mp.2016.48 27046646
Cadet JL, Brannock C, Krasnova IN, Jayanthi S, Ladenheim B, McCoy MT, et al. Genome-wide DNA hydroxymethylation identifies potassium channels in the nucleus accumbens as discriminators of methamphetamine addiction and abstinence. Mol Psychiatry. 2017;22:1196–204.27046646
29. Duan Y Tsai PJ Salmeron BJ Hu Y Gu H Lu H Compulsive drug-taking is associated with habenula-frontal cortex connectivity PNAS 2022 119 e2208867119 10.1073/pnas.2208867119 36469769
Duan Y, Tsai PJ, Salmeron BJ, Hu Y, Gu H, Lu H, et al. Compulsive drug-taking is associated with habenula-frontal cortex connectivity. PNAS. 2022;119:e2208867119.36469769
30. Hu Y Salmeron BJ Krasnova IN Gu H Lu H Bonci A Compulsive drug use is associated with imbalance of orbitofrontal- and prelimbic-striatal circuits in punishment-resistant individuals Proc Natl Acad Sci USA 2019 116 9066 71 10.1073/pnas.1819978116 30988198
Hu Y, Salmeron BJ, Krasnova IN, Gu H, Lu H, Bonci A, et al. Compulsive drug use is associated with imbalance of orbitofrontal- and prelimbic-striatal circuits in punishment-resistant individuals. Proc Natl Acad Sci USA. 2019;116:9066–71.30988198
31. Jayanthi S Ladenheim B Sullivan P McCoy MT Krasnova IN Goldstein DS Biochemical neuroadaptations in the rat striatal dopaminergic system after prolonged exposure to methamphetamine self-administration Int J Mol Sci 2022 23 10092 10.3390/ijms231710092 36077488
Jayanthi S, Ladenheim B, Sullivan P, McCoy MT, Krasnova IN, Goldstein DS, et al. Biochemical neuroadaptations in the rat striatal dopaminergic system after prolonged exposure to methamphetamine self-administration. Int J Mol Sci. 2022;23:10092.36077488
32. Munoz C Jayanthi S Ladenheim B Cadet JL Compulsive methamphetamine self-administration in the presence of adverse consequences is associated with increased hippocampal mRNA expression of cellular adhesion molecules Front Mol Neurosci 2023 15 1104657 10.3389/fnmol.2022.1104657 36710935
Munoz C, Jayanthi S, Ladenheim B, Cadet JL. Compulsive methamphetamine self-administration in the presence of adverse consequences is associated with increased hippocampal mRNA expression of cellular adhesion molecules. Front Mol Neurosci. 2023;15:1104657.36710935
33. Torres OV Jayanthi S Ladenheim B McCoy MT Krasnova IN Cadet JL Compulsive methamphetamine taking under punishment is associated with greater cue-induced drug seeking in rats Behav Brain Res 2017 326 265 71 10.1016/j.bbr.2017.03.009 28284948
Torres OV, Jayanthi S, Ladenheim B, McCoy MT, Krasnova IN, Cadet JL. Compulsive methamphetamine taking under punishment is associated with greater cue-induced drug seeking in rats. Behav Brain Res. 2017;326:265–71.28284948
34. Hu Y Salmeron BJ Gu H Stein EA Yang Y Impaired functional connectivity within and between frontostriatal circuits and its association with compulsive drug use and trait impulsivity in cocaine addiction JAMA Psychiatry 2015 72 584 92 10.1001/jamapsychiatry.2015.1 25853901
Hu Y, Salmeron BJ, Gu H, Stein EA, Yang Y. Impaired functional connectivity within and between frontostriatal circuits and its association with compulsive drug use and trait impulsivity in cocaine addiction. JAMA Psychiatry. 2015;72:584–92.25853901
35. Ceceli AO Bradberry CW Goldstein RZ The neurobiology of drug addiction: Cross-species insights into the dysfunction and recovery of the prefrontal cortex Neuropsychopharmacology 2022 47 276 91 10.1038/s41386-021-01153-9 34408275
Ceceli AO, Bradberry CW, Goldstein RZ. The neurobiology of drug addiction: Cross-species insights into the dysfunction and recovery of the prefrontal cortex. Neuropsychopharmacology. 2022;47:276–91.34408275
36. Luciana M Collins PF Neuroplasticity, the prefrontal cortex, and psychopathology-related deviations in cognitive control Annu Rev Clin Psychol 2022 18 443 69 10.1146/annurev-clinpsy-081219-111203 35534121
Luciana M, Collins PF. Neuroplasticity, the prefrontal cortex, and psychopathology-related deviations in cognitive control. Annu Rev Clin Psychol. 2022;18:443–69.35534121
37. Torres OV Jayanthi S McCoy MT Cadet JL Selective activation of striatal NGF-TrkA/p75NTR/MAPK intracellular signaling in rats that show suppression of methamphetamine intake 30 days following drug abstinence Int J Neuropsychopharmacol 2018 21 281 90 10.1093/ijnp/pyx105 29165617
Torres OV, Jayanthi S, McCoy MT, Cadet JL. Selective activation of striatal NGF-TrkA/p75NTR/MAPK intracellular signaling in rats that show suppression of methamphetamine intake 30 days following drug abstinence. Int J Neuropsychopharmacol. 2018;21:281–90.29165617
38. Campbell RR Chen S Beardwood JH López AJ Pham LV Keiser AM Cocaine induces paradigm-specific changes to the transcriptome within the ventral tegmental area Neuropsychopharmacology 2021 46 1768 79 10.1038/s41386-021-01031-4 34155331
Campbell RR, Chen S, Beardwood JH, López AJ, Pham LV, Keiser AM, et al. Cocaine induces paradigm-specific changes to the transcriptome within the ventral tegmental area. Neuropsychopharmacology. 2021;46:1768–79.34155331
39. Mews P Cunningham AM Scarpa J Ramakrishnan A Hicks EM Bolnick S Convergent abnormalities in striatal gene networks in human cocaine use disorder and mouse cocaine administration models Sci Adv 2023 9 eadd8946 10.1126/sciadv.add8946 36763659
Mews P, Cunningham AM, Scarpa J, Ramakrishnan A, Hicks EM, Bolnick S, et al. Convergent abnormalities in striatal gene networks in human cocaine use disorder and mouse cocaine administration models. Sci Adv. 2023;9:eadd8946.36763659
40. Mayberry HL Bavley CC Karbalaei R Peterson DR Bongiovanni AR Ellis AS Transcriptomics in the nucleus accumbens shell reveal sex- and reinforcer-specific signatures associated with morphine and sucrose craving Neuropsychopharmacology 2022 47 1764 75 10.1038/s41386-022-01289-2 35190706
Mayberry HL, Bavley CC, Karbalaei R, Peterson DR, Bongiovanni AR, Ellis AS, et al. Transcriptomics in the nucleus accumbens shell reveal sex- and reinforcer-specific signatures associated with morphine and sucrose craving. Neuropsychopharmacology. 2022;47:1764–75.35190706
41. Mansouri S Pessoni AM Marroquín-Rivera A Parise EM Tamminga CA Turecki G Transcriptional dissection of symptomatic profiles across the brain of men and women with depression Nat Commun 2023 14 6835 10.1038/s41467-023-42686-5 37884562
Mansouri S, Pessoni AM, Marroquín-Rivera A, Parise EM, Tamminga CA, Turecki G, et al. Transcriptional dissection of symptomatic profiles across the brain of men and women with depression. Nat Commun. 2023;14:6835.37884562
42. Blackwood CA Leary M Salisbury A McCoy MT Cadet JL Escalated oxycodone self-administration causes differential striatal mRNA expression of FGFs and IEGs following abstinence-associated incubation of oxycodone craving Neuroscience 2019 415 173 83 10.1016/j.neuroscience.2019.07.030 31351142
Blackwood CA, Leary M, Salisbury A, McCoy MT, Cadet JL. Escalated oxycodone self-administration causes differential striatal mRNA expression of FGFs and IEGs following abstinence-associated incubation of oxycodone craving. Neuroscience. 2019;415:173–83.31351142
43. Daiwile AP Jayanthi S Ladenheim B McCoy MT Brannock C Schroeder J Sex differences in escalated methamphetamine self-administration and altered gene expression associated with incubation of methamphetamine seeking Int J Neuropsychopharmacol 2019 22 710 23 10.1093/ijnp/pyz050 31562746
Daiwile AP, Jayanthi S, Ladenheim B, McCoy MT, Brannock C, Schroeder J, et al. Sex differences in escalated methamphetamine self-administration and altered gene expression associated with incubation of methamphetamine seeking. Int J Neuropsychopharmacol. 2019;22:710–23.31562746
44. Cadet JL Krasnova IN Walther D Brannock C Ladenheim B McCoy MT Increased expression of proenkephalin and prodynorphin mRNAs in the nucleus accumbens of compulsive methamphetamine taking rats Sci Rep 2016 6 37002 10.1038/srep37002 27841313
Cadet JL, Krasnova IN, Walther D, Brannock C, Ladenheim B, McCoy MT, et al. Increased expression of proenkephalin and prodynorphin mRNAs in the nucleus accumbens of compulsive methamphetamine taking rats. Sci Rep. 2016;6:37002.27841313
45. Jayanthi S Gonzalez B McCoy MT Ladenheim B Bisagno V Cadet JL Methamphetamine induces TET1- and TET3-dependent DNA hydroxymethylation of Crh and Avp genes in the rat nucleus accumbens Mol Neurobiol 2018 55 5154 66 10.1007/s12035-017-0750-9 28842817
Jayanthi S, Gonzalez B, McCoy MT, Ladenheim B, Bisagno V, Cadet JL. Methamphetamine induces TET1- and TET3-dependent DNA hydroxymethylation of Crh and Avp genes in the rat nucleus accumbens. Mol Neurobiol. 2018;55:5154–66.28842817
46. Giuliano C Peña-Oliver Y Goodlett CR Cardinal RN Robbins TW Bullmore ET Evidence for a long-lasting compulsive alcohol seeking phenotype in rats Neuropsychopharmacology 2018 43 728 38 10.1038/npp.2017.105 28553834
Giuliano C, Peña-Oliver Y, Goodlett CR, Cardinal RN, Robbins TW, Bullmore ET, et al. Evidence for a long-lasting compulsive alcohol seeking phenotype in rats. Neuropsychopharmacology. 2018;43:728–38.28553834
47. Grimm JW Hope BT Wise RA Shaham Y Neuroadaptation. incubation of cocaine craving after withdrawal Nature 2001 412 141 2 10.1038/35084134 11449260
Grimm JW, Hope BT, Wise RA, Shaham Y. Neuroadaptation. incubation of cocaine craving after withdrawal. Nature. 2001;412:141–2.11449260
48. Fredriksson I Venniro M Reiner DJ Chow JJ Bossert JM Shaham Y Animal models of drug relapse and craving after voluntary abstinence: a review Pharmacol Rev 2021 73 1050 83 10.1124/pharmrev.120.000191 34257149
Fredriksson I, Venniro M, Reiner DJ, Chow JJ, Bossert JM, Shaham Y. Animal models of drug relapse and craving after voluntary abstinence: a review. Pharmacol Rev. 2021;73:1050–83.34257149
49. Madangopal R Szelenyi ER Nguyen J Brenner MB Drake OR Pham DQ Incubation of palatable food craving is associated with brain-wide neuronal activation in mice PNAS 2022 119 e2209382119 10.1073/pnas.2209382119 36603188
Madangopal R, Szelenyi ER, Nguyen J, Brenner MB, Drake OR, Pham DQ, et al. Incubation of palatable food craving is associated with brain-wide neuronal activation in mice. PNAS. 2022;119:e2209382119.36603188
50. Wang G Shi J Chen N Xu L Li J Li P Effects of length of abstinence on decision-making and craving in methamphetamine abusers PLoS One 2013 8 e68791 10.1371/journal.pone.0068791 23894345
Wang G, Shi J, Chen N, Xu L, Li J, Li P, et al. Effects of length of abstinence on decision-making and craving in methamphetamine abusers. PLoS One. 2013;8:e68791.23894345
51. Altshuler RD Lin H Li X Neural mechanisms underlying incubation of methamphetamine craving: a mini-review Pharmacol Biochem Behav 2020 199 173058 10.1016/j.pbb.2020.173058 33250444
Altshuler RD, Lin H, Li X. Neural mechanisms underlying incubation of methamphetamine craving: a mini-review. Pharmacol Biochem Behav. 2020;199:173058.33250444
52. Daiwile AP Jayanthi S Cadet JL Sex- and brain region-specific changes in gene expression in male and female rats as consequences of methamphetamine self-administration and abstinence Neuroscience 2021 452 265 79 10.1016/j.neuroscience.2020.11.025 33242543
Daiwile AP, Jayanthi S, Cadet JL. Sex- and brain region-specific changes in gene expression in male and female rats as consequences of methamphetamine self-administration and abstinence. Neuroscience. 2021;452:265–79.33242543
53. Bernhardt N Petzold J Groß C Scheck A Pooseh S Mayer-Pelinski R Neurocognitive dysfunctions and their therapeutic modulation in patients with methamphetamine dependence: a pilot study Front Psychiatry 2020 11 581 10.3389/fpsyt.2020.00581 32714215
Bernhardt N, Petzold J, Groß C, Scheck A, Pooseh S, Mayer-Pelinski R, et al. Neurocognitive dysfunctions and their therapeutic modulation in patients with methamphetamine dependence: a pilot study. Front Psychiatry. 2020;11:581.32714215
54. French L Ma T Oh H Tseng GC Sibille E Age-related gene expression in the frontal cortex suggests synaptic function changes in specific inhibitory neuron subtypes Front Aging Neurosci 2017 9 162 10.3389/fnagi.2017.00162 28611654
French L, Ma T, Oh H, Tseng GC, Sibille E. Age-related gene expression in the frontal cortex suggests synaptic function changes in specific inhibitory neuron subtypes. Front Aging Neurosci. 2017;9:162.28611654
55. Castrén E Neurotrophins as mediators of drug effects on mood, addiction, and neuroprotection Mol Neurobiol 2004 29 289 302 10.1385/MN:29:3:289 15181240
Castrén E. Neurotrophins as mediators of drug effects on mood, addiction, and neuroprotection. Mol Neurobiol. 2004;29:289–302.15181240
56. Liran M Rahamim N Ron D Barak S Growth factors and alcohol use disorder Cold Spring Harb Perspect Med 2020 10 a039271 10.1101/cshperspect.a039271 31964648
Liran M, Rahamim N, Ron D, Barak S. Growth factors and alcohol use disorder. Cold Spring Harb Perspect Med. 2020;10:a039271.31964648
57. Ghitza UE Zhai H Wu P Airavaara M Shaham Y Lu L Role of BDNF and GDNF in drug reward and relapse: a review Neurosci Biobehav Rev 2010 35 157 71 10.1016/j.neubiorev.2009.11.009 19914287
Ghitza UE, Zhai H, Wu P, Airavaara M, Shaham Y, Lu L. Role of BDNF and GDNF in drug reward and relapse: a review. Neurosci Biobehav Rev. 2010;35:157–71.19914287
58. Even-Chen O Barak S The role of fibroblast growth factor 2 in drug addiction Eur J Neurosci 2019 50 2552 61 10.1111/ejn.14133 30144335
Even-Chen O, Barak S. The role of fibroblast growth factor 2 in drug addiction. Eur J Neurosci. 2019;50:2552–61.30144335
59. Torres OV McCoy MT Ladenheim B Jayanthi S Brannock C Tulloch I CAMKII-conditional deletion of histone deacetylase 2 potentiates acute methamphetamine-induced expression of immediate early genes in the mouse nucleus accumbens Sci Rep 2015 5 13396 10.1038/srep13396 26300473
Torres OV, McCoy MT, Ladenheim B, Jayanthi S, Brannock C, Tulloch I, et al. CAMKII-conditional deletion of histone deacetylase 2 potentiates acute methamphetamine-induced expression of immediate early genes in the mouse nucleus accumbens. Sci Rep. 2015;5:13396.26300473
60. Torres OV Ladenheim B Jayanthi S McCoy MT Krasnova IN Vautier FA An acute methamphetamine injection downregulates the expression of several histone deacetylases (HDACs) in the mouse nucleus accumbens: Potential regulatory role of HDAC2 expression Neurotox Res 2016 30 32 40 10.1007/s12640-015-9591-3 26721795
Torres OV, Ladenheim B, Jayanthi S, McCoy MT, Krasnova IN, Vautier FA, et al. An acute methamphetamine injection downregulates the expression of several histone deacetylases (HDACs) in the mouse nucleus accumbens: Potential regulatory role of HDAC2 expression. Neurotox Res. 2016;30:32–40.26721795
61. Cass WA Gerhardt GA Gillespie K Curella P Mayfield RD Zahniser NR Reduced clearance of exogenous dopamine in rat nucleus accumbens, but not in dorsal striatum, following cocaine challenge in rats withdrawn from repeated cocaine administration J Neurochem 1993 61 273 83 10.1111/j.1471-4159.1993.tb03565.x 8515274
Cass WA, Gerhardt GA, Gillespie K, Curella P, Mayfield RD, Zahniser NR. Reduced clearance of exogenous dopamine in rat nucleus accumbens, but not in dorsal striatum, following cocaine challenge in rats withdrawn from repeated cocaine administration. J Neurochem. 1993;61:273–83.8515274
62. Chen BT Yau HJ Hatch C Kusumoto-Yoshida I Cho SL Hopf FW Rescuing cocaine-induced prefrontal cortex hypoactivity prevents compulsive cocaine seeking Nature 2013 496 359 62 10.1038/nature12024 23552889
Chen BT, Yau HJ, Hatch C, Kusumoto-Yoshida I, Cho SL, Hopf FW, et al. Rescuing cocaine-induced prefrontal cortex hypoactivity prevents compulsive cocaine seeking. Nature. 2013;496:359–62.23552889
63. Piazza PV Deminière JM Le Moal M Simon H Factors that predict individual vulnerability to amphetamine self-administration Science 1989 245 1511 3 10.1126/science.2781295 2781295
Piazza PV, Deminière JM, Le Moal M, Simon H. Factors that predict individual vulnerability to amphetamine self-administration. Science. 1989;245:1511–3.2781295
64. Sabeti J Gerhardt GA Zahniser NR Acute cocaine differentially alters accumbens and striatal dopamine clearance in low and high cocaine locomotor responders: behavioral and electrochemical recordings in freely moving rats J Pharmacol Exp Ther 2002 302 1201 11 10.1124/jpet.102.035816 12183681
Sabeti J, Gerhardt GA, Zahniser NR. Acute cocaine differentially alters accumbens and striatal dopamine clearance in low and high cocaine locomotor responders: behavioral and electrochemical recordings in freely moving rats. J Pharmacol Exp Ther. 2002;302:1201–11.12183681
65. Segal DS Kuczenski R Individual differences in responsiveness to single and repeated amphetamine administration: behavioral characteristics and neurochemical correlates J Pharmacol Exp Ther 1987 242 917 26 3656119
Segal DS, Kuczenski R. Individual differences in responsiveness to single and repeated amphetamine administration: behavioral characteristics and neurochemical correlates. J Pharmacol Exp Ther. 1987;242:917–26.3656119
66. Pelloux Y Everitt BJ Dickinson A Compulsive drug seeking by rats under punishment: effects of drug taking history Psychopharmacology (Berl) 2007 194 127 37 10.1007/s00213-007-0805-0 17514480
Pelloux Y, Everitt BJ, Dickinson A. Compulsive drug seeking by rats under punishment: effects of drug taking history. Psychopharmacology (Berl). 2007;194:127–37.17514480
67. Kelly TH Robbins G Martin CA Fillmore MT Lane SD Harrington NG Individual differences in drug abuse vulnerability: d-amphetamine and sensation-seeking status Psychopharmacology 2006 189 17 25 10.1007/s00213-006-0487-z 16972106
Kelly TH, Robbins G, Martin CA, Fillmore MT, Lane SD, Harrington NG, et al. Individual differences in drug abuse vulnerability: d-amphetamine and sensation-seeking status. Psychopharmacology. 2006;189:17–25.16972106
68. Harvanko A Martin C Lile J Kryscio R Kelly TH Individual differences in the reinforcing and subjective effects of d-amphetamine: Dimensions of impulsivity Exp Clin Psychopharmacol 2016 24 436 46 10.1037/pha0000095 27929346
Harvanko A, Martin C, Lile J, Kryscio R, Kelly TH. Individual differences in the reinforcing and subjective effects of d-amphetamine: Dimensions of impulsivity. Exp Clin Psychopharmacol. 2016;24:436–46.27929346
69. Lattal KA Cançado CRX Cook JE Kincaid SL Nighbor TD Oliver AC On defining resurgence Behav Processes 2017 141 85 91 10.1016/j.beproc.2017.04.018 28487201
Lattal KA, Cançado CRX, Cook JE, Kincaid SL, Nighbor TD, Oliver AC. On defining resurgence. Behav Processes. 2017;141:85–91.28487201
70. Nall RW Rung JM Shahan TA Resurgence of a target behavior suppressed by a combination of punishment and alternative reinforcement Behav Processes 2019 162 177 83 10.1016/j.beproc.2019.03.004 30862521
Nall RW, Rung JM, Shahan TA. Resurgence of a target behavior suppressed by a combination of punishment and alternative reinforcement. Behav Processes. 2019;162:177–83.30862521
71. Bolívar HA Dallery J Effects of response cost magnitude on resurgence of human operant behavior Behav Proces 2020 178 104187 10.1016/j.beproc.2020.104187
Bolívar HA, Dallery J. Effects of response cost magnitude on resurgence of human operant behavior. Behav Proces. 2020;178:104187.
72. Podlesnik CA Kuroda T Jimenez-Gomez C Abreu-Rodrigues J Cançado CRX Blackman AL Resurgence is greater following a return to the training context than remaining in the extinction context J Exp Anal Behav 2019 111 416 35 10.1002/jeab.505 30706484
Podlesnik CA, Kuroda T, Jimenez-Gomez C, Abreu-Rodrigues J, Cançado CRX, Blackman AL, et al. Resurgence is greater following a return to the training context than remaining in the extinction context. J Exp Anal Behav. 2019;111:416–35.30706484
73. Podlesnik CA Ritchey CM Waits J Gilroy SP A Comprehensive Systematic Review of Procedures and Analyses Used in Basic and Preclinical Studies of Resurgence, 1970-2020 Perspect Behav Sci 2022 46 137 84 10.1007/s40614-022-00361-y 37006602
Podlesnik CA, Ritchey CM, Waits J, Gilroy SP. A Comprehensive Systematic Review of Procedures and Analyses Used in Basic and Preclinical Studies of Resurgence, 1970-2020. Perspect Behav Sci. 2022;46:137–84.37006602
74. Alessandri J Cançado CRX Context affects resurgence of negatively reinforced human behavior Behav Process 2020 170 104018 10.1016/j.beproc.2019.104018
Alessandri J, Cançado CRX. Context affects resurgence of negatively reinforced human behavior. Behav Process. 2020;170:104018.
75. Kashani JH Solomon NA Dugan K Joy F Differences between early and late onset of substance abuse: an inpatient experience South Med J 1987 80 554 7 10.1097/00007611-198705000-00003 3576265
Kashani JH, Solomon NA, Dugan K, Joy F. Differences between early and late onset of substance abuse: an inpatient experience. South Med J. 1987;80:554–7.3576265
76. Drossel G Brucar LR Rawls E Hendrickson TJ Zilverstand A Subtypes in addiction and their neurobehavioral profiles across three functional domains Transl Psychiatry 2023 13 127 10.1038/s41398-023-02426-1 37072391
Drossel G, Brucar LR, Rawls E, Hendrickson TJ, Zilverstand A. Subtypes in addiction and their neurobehavioral profiles across three functional domains. Transl Psychiatry. 2023;13:127.37072391
77. Cloninger CR Neurogenetic adaptive mechanisms in alcoholism Science 1987 236 410 6 10.1126/science.2882604 2882604
Cloninger CR. Neurogenetic adaptive mechanisms in alcoholism. Science. 1987;236:410–6.2882604
78. Ball SA Carroll KM Babor TF Rounsaville BJ Subtypes of cocaine abusers: support for a type A-type B distinction J Consult Clin Psychol 1995 63 115 24 10.1037/0022-006X.63.1.115 7896976
Ball SA, Carroll KM, Babor TF, Rounsaville BJ. Subtypes of cocaine abusers: support for a type A-type B distinction. J Consult Clin Psychol. 1995;63:115–24.7896976
79. Debeck K Kerr T Marshall BD Simo A Montaner J Wood E Risk factors for progression to regular injection drug use among street-involved youth in a Canadian setting Drug Alcohol Depend 2013 133 468 72 10.1016/j.drugalcdep.2013.07.008 23910434
Debeck K, Kerr T, Marshall BD, Simo A, Montaner J, Wood E. Risk factors for progression to regular injection drug use among street-involved youth in a Canadian setting. Drug Alcohol Depend. 2013;133:468–72.23910434
80. Wathen SN Podlesnik CA Laboratory models of treatment relapse and mitigation techniques Behav Anal: Res Pract 2018 18 362 87
Wathen SN, Podlesnik CA. Laboratory models of treatment relapse and mitigation techniques. Behav Anal: Res Pract. 2018;18:362–87.
81. Aziz A Harrop SP Bishop NE Characterization of the deleted in autism 1 protein family: implications for studying cognitive disorders PLoS One 2011 6 e14547 10.1371/journal.pone.0014547 21283809
Aziz A, Harrop SP, Bishop NE. Characterization of the deleted in autism 1 protein family: implications for studying cognitive disorders. PLoS One. 2011;6:e14547.21283809
82. Aziz A Harrop SP Bishop NE DIA1R is an X-linked gene related to deleted in Autism-1 PLoS One 2011 6 e14534 10.1371/journal.pone.0014534 21264219
Aziz A, Harrop SP, Bishop NE. DIA1R is an X-linked gene related to deleted in Autism-1. PLoS One. 2011;6:e14534.21264219
83. Tsai CH Cheng HC Wang YS Lin P Jen J Kuo IY Small GTPase Rab37 targets tissue inhibitor of metalloproteinase 1 for exocytosis and thus suppresses tumour metastasis Nat Commun 2014 5 4804 10.1038/ncomms5804 25183545
Tsai CH, Cheng HC, Wang YS, Lin P, Jen J, Kuo IY, et al. Small GTPase Rab37 targets tissue inhibitor of metalloproteinase 1 for exocytosis and thus suppresses tumour metastasis. Nat Commun. 2014;5:4804.25183545
84. Caldwell HK Lee HJ Macbeth AH Young WS 3rd Vasopressin: behavioral roles of an “original” neuropeptide Prog Neurobiol 2008 84 1 24 10.1016/j.pneurobio.2007.10.007 18053631
Caldwell HK, Lee HJ, Macbeth AH, Young WS 3rd. Vasopressin: behavioral roles of an “original” neuropeptide. Prog Neurobiol. 2008;84:1–24.18053631
85. Raggenbass M Overview of cellular electrophysiological actions of vasopressin Eur J Pharmacol 2008 583 243 54 10.1016/j.ejphar.2007.11.074 18280467
Raggenbass M. Overview of cellular electrophysiological actions of vasopressin. Eur J Pharmacol. 2008;583:243–54.18280467
86. Bisagno V Cadet JL Stress, sex, and addiction: potential roles of corticotropin-releasing factor, oxytocin, and arginine-vasopressin Behav Pharmacol 2014 25 445 57 10.1097/FBP.0000000000000049 24949572
Bisagno V, Cadet JL. Stress, sex, and addiction: potential roles of corticotropin-releasing factor, oxytocin, and arginine-vasopressin. Behav Pharmacol. 2014;25:445–57.24949572
87. Cadet JL Brannock C Ladenheim B McCoy MT Krasnova IN Lehrmann E Enhanced upregulation of CRH mRNA expression in the nucleus accumbens of male rats after a second injection of methamphetamine given thirty days later PLoS One 2014 9 e84665 10.1371/journal.pone.0084665 24475032
Cadet JL, Brannock C, Ladenheim B, McCoy MT, Krasnova IN, Lehrmann E, et al. Enhanced upregulation of CRH mRNA expression in the nucleus accumbens of male rats after a second injection of methamphetamine given thirty days later. PLoS One. 2014;9:e84665.24475032
88. Zhou Y Leri F Cummins E Hoeschele M Kreek MJ Involvement of arginine vasopressin and V1b receptor in heroin withdrawal and heroin seeking precipitated by stress and by heroin Neuropsychopharmacology 2008 33 226 36 10.1038/sj.npp.1301419 17443128
Zhou Y, Leri F, Cummins E, Hoeschele M, Kreek MJ. Involvement of arginine vasopressin and V1b receptor in heroin withdrawal and heroin seeking precipitated by stress and by heroin. Neuropsychopharmacology. 2008;33:226–36.17443128
89. Flores C Samaha AN Stewart J Requirement of endogenous basic fibroblast growth factor for sensitization to amphetamine J Neurosci 2000 20 RC55 10.1523/JNEUROSCI.20-02-j0003.2000 10632621
Flores C, Samaha AN, Stewart J. Requirement of endogenous basic fibroblast growth factor for sensitization to amphetamine. J Neurosci. 2000;20:RC55.10632621
90. Yin W Clare K Zhang Q Volkow ND Du C Chronic cocaine induces HIF-VEGF pathway activation along with angiogenesis in the brain PLoS One 2017 12 e0175499 10.1371/journal.pone.0175499 28448515
Yin W, Clare K, Zhang Q, Volkow ND, Du C. Chronic cocaine induces HIF-VEGF pathway activation along with angiogenesis in the brain. PLoS One. 2017;12:e0175499.28448515
91. Kim YK Lee BC Ham BJ Yang BH Roh S Choi J Increased transforming growth factor-beta1 in alcohol dependence J Korean Med Sci 2009 24 941 4 10.3346/jkms.2009.24.5.941 19794996
Kim YK, Lee BC, Ham BJ, Yang BH, Roh S, Choi J, et al. Increased transforming growth factor-beta1 in alcohol dependence. J Korean Med Sci. 2009;24:941–4.19794996
92. Morimoto Y Yoshida S Kinoshita A Satoh C Mishima H Yamaguchi N Nonsense mutation in CFAP43 causes normal-pressure hydrocephalus with ciliary abnormalities Neurology 2019 92 e2364 e2374 10.1212/WNL.0000000000007505 31004071
Morimoto Y, Yoshida S, Kinoshita A, Satoh C, Mishima H, Yamaguchi N, et al. Nonsense mutation in CFAP43 causes normal-pressure hydrocephalus with ciliary abnormalities. Neurology. 2019;92:e2364–e2374.31004071
93. Chen XJ Xu H Cooper HM Liu Y Cytoplasmic dynein: a key player in neurodegenerative and neurodevelopmental diseases Sci China Life Sci 2014 57 372 7 10.1007/s11427-014-4639-9 24664850
Chen XJ, Xu H, Cooper HM, Liu Y. Cytoplasmic dynein: a key player in neurodegenerative and neurodevelopmental diseases. Sci China Life Sci. 2014;57:372–7.24664850
94. Moughamian AJ, Holzbaur ELF Chapter 13 - Cytoplasmic dynein dysfunction and neurodegenerative disease. Dyneins: structure, biology and disease. 2nd ed. Academic Press; 2018. p. 286–315. ISBN 9780128094709. 10.1016/B978-0-12-809470-9.00013-8.
95. McCoy MT Jayanthi S Cadet JL Potassium channels and their potential roles in substance use disorders Int J Mol Sci 2021 22 1249 10.3390/ijms22031249 33513859
McCoy MT, Jayanthi S, Cadet JL. Potassium channels and their potential roles in substance use disorders. Int J Mol Sci. 2021;22:1249.33513859
96. Luo Y Liao C Chen L Zhang Y Bao S Deng A Heroin addiction induces axonal transport dysfunction in the brain detected by in vivo MRI Neurotox Res 2022 40 1070 85 10.1007/s12640-022-00533-3 35759084
Luo Y, Liao C, Chen L, Zhang Y, Bao S, Deng A, et al. Heroin addiction induces axonal transport dysfunction in the brain detected by in vivo MRI. Neurotox Res. 2022;40:1070–85.35759084
97. Martin TA Jayanthi S McCoy MT Brannock C Ladenheim B Garrett T Methamphetamine causes differential alterations in gene expression and patterns of histone acetylation/hypoacetylation in the rat nucleus accumbens PLoS One 2012 7 e34236 10.1371/journal.pone.0034236 22470541
Martin TA, Jayanthi S, McCoy MT, Brannock C, Ladenheim B, Garrett T, et al. Methamphetamine causes differential alterations in gene expression and patterns of histone acetylation/hypoacetylation in the rat nucleus accumbens. PLoS One. 2012;7:e34236.22470541
98. Browne CJ Godino A Salery M Nestler EJ Epigenetic mechanisms of opioid addiction Biol Psychiatry 2020 87 22 33 10.1016/j.biopsych.2019.06.027 31477236
Browne CJ, Godino A, Salery M, Nestler EJ. Epigenetic mechanisms of opioid addiction. Biol Psychiatry. 2020;87:22–33.31477236
99. Jayanthi S McCoy MT Cadet JL Epigenetic regulatory dynamics in models of methamphetamine-use disorder Genes 2021 12 1614 10.3390/genes12101614 34681009
Jayanthi S, McCoy MT, Cadet JL. Epigenetic regulatory dynamics in models of methamphetamine-use disorder. Genes. 2021;12:1614.34681009
100. Poisel E Zillich L Streit F Frank J Friske MM Foo JC DNA methylation in cocaine use disorder-An epigenome-wide approach in the human prefrontal cortex Front Psychiatry 2023 14 1075250 10.3389/fpsyt.2023.1075250 36865068
Poisel E, Zillich L, Streit F, Frank J, Friske MM, Foo JC, et al. DNA methylation in cocaine use disorder-An epigenome-wide approach in the human prefrontal cortex. Front Psychiatry. 2023;14:1075250.36865068
101. Pao PC Tsai LH Histone deacetylases 1 and 2 in memory function ACS Chem Neurosci 2022 13 848 58 10.1021/acschemneuro.1c00775 35263084
Pao PC, Tsai LH. Histone deacetylases 1 and 2 in memory function. ACS Chem Neurosci. 2022;13:848–58.35263084
102. Bisagno V Cadet JL Histone deacetylases and immediate early genes: key players in psychostimulant-induced neuronal plasticity Neurotox Res 2021 39 2134 40 10.1007/s12640-021-00420-3 34581974
Bisagno V, Cadet JL. Histone deacetylases and immediate early genes: key players in psychostimulant-induced neuronal plasticity. Neurotox Res. 2021;39:2134–40.34581974
103. Torres OV Sex differences in psychostimulant abuse: implications for estrogen receptors and histone deacetylases Genes 2022 13 892 10.3390/genes13050892 35627277
Torres OV. Sex differences in psychostimulant abuse: implications for estrogen receptors and histone deacetylases. Genes. 2022;13:892.35627277
104. Guan JS Haggarty SJ Giacometti E Dannenberg JH Joseph N Gao J HDAC2 negatively regulates memory formation and synaptic plasticity Nature 2009 459 55 60 10.1038/nature07925 19424149
Guan JS, Haggarty SJ, Giacometti E, Dannenberg JH, Joseph N, Gao J, et al. HDAC2 negatively regulates memory formation and synaptic plasticity. Nature. 2009;459:55–60.19424149
105. Graham DL Edwards S Bachtell RK DiLeone RJ Rios M Self DW Dynamic BDNF activity in nucleus accumbens with cocaine use increases self-administration and relapse Nature Neurosci 2007 10 1029 37 10.1038/nn1929 17618281
Graham DL, Edwards S, Bachtell RK, DiLeone RJ, Rios M, Self DW. Dynamic BDNF activity in nucleus accumbens with cocaine use increases self-administration and relapse. Nature Neurosci. 2007;10:1029–37.17618281
106. St Laurent R Helm SR Glenn MJ Reduced cocaine-seeking behavior in heterozygous BDNF knockout rats Neurosci Lett 2013 544 94 99 10.1016/j.neulet.2013.03.050 23583595
St Laurent R, Helm SR, Glenn MJ. Reduced cocaine-seeking behavior in heterozygous BDNF knockout rats. Neurosci Lett. 2013;544:94–99.23583595
107. Morris MJ Mahgoub M Na ES Pranav H Monteggia LM Loss of histone deacetylase 2 improves working memory and accelerates extinction learning J Neurosci 2013 33 6401 11 10.1523/JNEUROSCI.1001-12.2013 23575838
Morris MJ, Mahgoub M, Na ES, Pranav H, Monteggia LM. Loss of histone deacetylase 2 improves working memory and accelerates extinction learning. J Neurosci. 2013;33:6401–11.23575838
