title: Incubation of methamphetamine craving is associated with selective increases in expression of Bdnf and trkb, glutamate receptors, and epigenetic enzymes in cue-activated fos-expressing dorsal striatal neurons.
abstract: Cue-induced methamphetamine seeking progressively increases after withdrawal (incubation of methamphetamine craving), but the underlying mechanisms are largely unknown. We determined whether this incubation is associated with alterations in candidate genes in dorsal striatum (DS), a brain area implicated in cue- and context-induced drug relapse. We first measured mRNA expression of 24 candidate genes in whole DS extracts after short (2 d) or prolonged (1 month) withdrawal in rats following extended-access methamphetamine or saline (control condition) self-administration (9 h/d, 10 d). We found minimal changes. Next, using fluorescence-activated cell sorting, we compared gene expression in Fos-positive dorsal striatal neurons, which were activated during "incubated" cue-induced drug-seeking tests after prolonged withdrawal, with nonactivated Fos-negative neurons. We found significant increases in mRNA expression of immediate early genes (Arc, Egr1), Bdnf and its receptor (Trkb), glutamate receptor subunits (Gria1, Gria3, Grm1), and epigenetic enzymes (Hdac3, Hdac4, Hdac5, GLP, Dnmt3a, Kdm1a) in the Fos-positive neurons only. Using RNAscope to determine striatal subregion and cell-type specificity of the activated neurons, we measured colabeling of Fos with Drd1 and Drd2 in three DS subregions. Fos expression was neither subregion nor cell-type specific (52.5 and 39.2% of Fos expression colabeled with Drd1 and Drd2, respectively). Finally, we found that DS injections of SCH23390 (C17H18ClNO), a D1-family receptor antagonist known to block cue-induced Fos induction, decreased incubated cue-induced methamphetamine seeking after prolonged withdrawal. Results demonstrate a critical role of DS in incubation of methamphetamine craving and that this incubation is associated with selective gene-expression alterations in cue-activated D1- and D2-expressing DS neurons.
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224, anna.li@nih.gov yavin.shaham@nih.gov.
authors.forename: Xuan
authors.identifier: http://orcid.org/0000-0002-6737-1223
authors.initials: X
authors.lastname: Li
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224.
authors.forename: F Javier
authors.initials: FJ
authors.lastname: Rubio
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224.
authors.forename: Tamara
authors.initials: T
authors.lastname: Zeric
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224.
authors.forename: Jennifer M
authors.identifier: http://orcid.org/0000-0001-5636-4071
authors.initials: JM
authors.lastname: Bossert
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224.
authors.forename: Sarita
authors.initials: S
authors.lastname: Kambhampati
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Hannah M
authors.identifier: http://orcid.org/0000-0002-9413-0847
authors.initials: HM
authors.lastname: Cates
authors.affiliation: Department of Psychology, University of California Los Angeles, Los Angeles, California 90095, and.
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: Kennedy
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224.
authors.forename: Qing-Rong
authors.initials: QR
authors.lastname: Liu
authors.affiliation: Division of Rheumatology, School of Medicine, Johns Hopkins University, Baltimore, Maryland 21224.
authors.forename: Raffaello
authors.initials: R
authors.lastname: Cimbro
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224.
authors.forename: Bruce T
authors.initials: BT
authors.lastname: Hope
authors.affiliation: Fishberg Department of Neuroscience and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, New York 10029.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Nestler
authors.affiliation: Behavioral Neuroscience Research Branch, Intramural Research Program, National Institute on Drug Abuse, National Institutes of Health, U.S. Department of Health and Human Services, Baltimore, Maryland 21224, anna.li@nih.gov yavin.shaham@nih.gov.
authors.forename: Yavin
authors.identifier: http://orcid.org/0000-0002-8242-3319
authors.initials: Y
authors.lastname: Shaham
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000096:biosynthesis*; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism*; D066249:Craving / Q000187:drug effects / Q000502:physiology*; D003463:Cues; D044127:Epigenesis, Genetic / Q000187:drug effects / Q000502:physiology; D005786:Gene Expression Regulation; D008297:Male; D008694:Methamphetamine / Q000008:administration & dosage*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D020813:Receptor, trkB / Q000096:biosynthesis*; D017470:Receptors, Glutamate / Q000096:biosynthesis*; D012646:Self Administration
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D016760:Proto-Oncogene Proteins c-fos; D017470:Receptors, Glutamate; D008694:Methamphetamine; D020813:Receptor, trkB
keywords: FACS; dorsal striatum; epigenetics; glutamate; methamphetamine; relapse