Fosb Regulates Wheel Running
Total Page：16
File Type：pdf, Size：1020Kb
The Journal of Neuroscience, September 15, 2002, 22(18):8133–8138 ⌬FosB Regulates Wheel Running Martin Werme,1 Chad Messer,3 Lars Olson,1 Lauren Gilden,3 Peter Thore´n,2 Eric J. Nestler,3 and Stefan Brene´ 1 Departments of 1Neuroscience and 2Physiology and Pharmacology, Karolinska Institutet, Stockholm, S-171 77 Sweden, and 3Department of Psychiatry and Center for Basic Neuroscience, The University of Texas Southwestern Medical Center, Dallas, Texas 75390-9070 ⌬FosB is a transcription factor that accumulates in a region- whereas mice that overexpress ⌬FosB predominantly in striatal specific manner in the brain after chronic perturbations. For enkephalin-containing neurons ran considerably less than con- example, repeated administration of drugs of abuse increases trols. Data from the present study demonstrate that like drugs levels of ⌬FosB in the striatum. In the present study, we ana- of abuse, voluntary running increases levels of ⌬FosB in brain lyzed the effect of spontaneous wheel running, as a model for a reward pathways. Furthermore, overexpression of ⌬FosB in a natural rewarding behavior, on levels of ⌬FosB in striatal re- distinct striatal output neuronal population increases running gions. Moreover, mice that inducibly overexpress ⌬FosB in behavior. Because previous work has shown that ⌬FosB over- specific subpopulations of striatal neurons were used to study expression within this same neuronal population increases the the possible role of ⌬FosB on running behavior. Lewis rats rewarding properties of drugs of abuse, results of the present given ad libitum access to running wheels for 30 d covered study suggest that ⌬FosB may play a key role in controlling what would correspond to ϳ10 km/d and showed increased both natural and drug-induced reward. levels of ⌬FosB in the nucleus accumbens compared with rats exposed to locked running wheels. Mice that overexpress Key words: nucleus accumbens; striatum; locomotion; exer- ⌬FosB selectively in striatal dynorphin-containing neurons in- cise; natural reward; behavioral addiction; compulsive; drugs of creased their daily running compared with control littermates, abuse ⌬FosB belongs to the Fos family of transcription factors and is 1998) to form long-lasting AP-1 complexes in specific brain re- derived from the fosb gene via alternative splicing. Unlike all gions. It has been proposed that these long-lasting AP-1 com- other Fos-like proteins, which have short half-lives, the 35 and 37 plexes mediate some of the long-term effects of drugs of abuse on kDa isoforms of ⌬FosB accumulate in a region-specific manner in brain reward pathways that underlie addiction (Nestler et al., the brain after a variety of chronic perturbations, presumably 2001). because of the very high stability of these isoforms (Hope et al., Behavioral studies suggest that wheel running in rodents is 1994a; Chen et al., 1997; Nestler et al., 1999). The regulation of rewarding. This assumption is based on experiments showing that ⌬ FosB in striatal regions after repeated administration of drugs of rats lever-press for access to running wheels and also develop abuse has been especially well studied (Hope et al., 1994b; Mor- conditioned place preference to an environment associated with atalla et al., 1996; Chen et al., 1997; Nestler et al., 1999). The the aftereffects of wheel running (Iversen, 1993; Belke, 1997; Lett mesolimbic dopamine pathway has a central role in drug reward et al., 2000). Moreover, rats that run long distances daily exhibit (Koob et al., 1998). It originates in the ventral tegmental area of withdrawal signs, such as increased aggression, when access to the the midbrain and terminates in the ventral part of the striatum, running wheels is denied (Hoffmann et al., 1987). Surveys among called the nucleus accumbens. Acute administration of any of highly committed human runners suggest that running is an several drugs of abuse transiently induces several Fos family addictive behavior for many individuals (Rudy and Estok, 1989; proteins in the nucleus accumbens and in the dorsal striatum. Chapman and De Castro, 1990; Furst and Germone, 1993). These proteins form heterodimers with Jun family proteins to Indeed, running displays many of the criteria included in the form activator protein-1 (AP-1) transcription factor complexes Diagnostic Statistical Manual (American Psychiatric Association, with short half-lives. In contrast, after repeated drug treatment, 1994) for the diagnosis of addiction. induction of these immediate early gene products declines and, ⌬ The goal of the present study was to investigate whether instead, there is a gradual accumulation of the stable FosB ⌬ isoforms. ⌬FosB heterodimerizes predominantly with JunD and levels of FosB are altered by a natural rewarding behavior to a lesser extent with JunB (Hiroi et al., 1998; Perez-Otano et al., such as running and whether inducible overexpression of ⌬FosB in striatal regions might regulate running behavior. We Received Jan. 29, 2002; revised June 11, 2002; accepted June 12, 2002. show here that, like drugs of abuse, chronic running induces This work was supported by the Swedish Research Council (03185, 11642, and ⌬FosB in the nucleus accumbens; in addition, the overexpres- 04762), the Centrum fo¨r idrottsforskning (CIF 86/01), the National Institute on ⌬ Drug Abuse, and the National Institute on Aging. We thank Karin Pernold and sion of FosB in the two different subsets of striatal projection Karin Lundstro¨mer for excellent technical assistance. neurons has opposite effects on wheel running. The data reveal Correspondence should be addressed to Stefan Brene´, Department of Neuro- striking similarities between addictive drugs and wheel running science, Karolinska Institutet, Stockholm, S-171 77 Sweden. E-mail: [email protected]. and suggest an important role for ⌬FosB in regulating both Copyright © 2002 Society for Neuroscience 0270-6474/02/228133-06$15.00/0 natural and drug-induced rewards. 8134 J. Neurosci., September 15, 2002, 22(18):8133–8138 Werme et al. • ⌬FosB Regulates Wheel Running MATERIALS AND METHODS section). This was followed by three rinses with PBS for 10 min before incubation for 1 hr at room temperature with the secondary biotinylated Animals. Male Lewis rats (Møllegaard Breeding Center, Skansved, Den- anti-rabbit IgG antibody (1:200; Vector Laboratories) in 0.3% Triton- mark) weighing 250 gm at the onset of the experiment were used. The PBS (150 l per section). Another three rinses in PBS for 10 min were rats had access ad libitum to water, food, and running wheels. They were performed before the avidin–biotin complex was added (1:100 and 1:100, on a 12 hr light/dark cycle, with lights on at 10 A.M. and lights off at 10 respectively, in 0.1 M PBS; 150 l per section). After three 10 min rinses, P.M. Cages (43 ϫ 22 ϫ 20 cm) contained a running wheel with a the complex was visualized aftera7minincubation with the substrate diameter of 34 cm; hence, one revolution corresponds to 1.07 m. After 4 according to the manufacturer’s protocol (Vector Laboratories). Sections weeks of voluntary wheel running, the rats were killed by decapitation, were subsequently rinsed three times for 5 min. and tissues were taken for Western blotting or perfused with fixative and In situ hybridization. For combined immunohistochemistry and in situ processed for immunohistochemistry and in situ hybridization. hybridization experiments, brain sections that had been processed for Two lines of bitransgenic mice that can inducibly overexpress ⌬FosB immunohistochemistry were immediately subjected to in situ hybridiza- selectively in striatal regions under the control of the tetracycline gene tion, which was performed essentially as described previously (Seroogy et regulation system were also used (Chen et al., 1998). In one line, called al., 1989; Dagerlind et al., 1992). Forty-eight mer DNA oligonucleotide 11A, ⌬FosB is inducibly overexpressed solely in striatal projection neu- probes specific for dynorphin (296–345) (Douglass et al., 1989) and rons that express the neuropeptide dynorphin after removal of doxycy- enkephalin (235–282) (Zurawski et al., 1986) mRNAs were radioactively cline (Kelz et al., 1999). In the other line, called 11B, ⌬FosB is inducibly labeled with [␣- 35S]dATP (DuPont NEN) in their 3Ј ends using terminal overexpressed predominantly in striatal projection neurons that express deoxynucleotidyl transferase (Invitrogen, San Diego, CA) to a specific the neuropeptide enkephalin after removal of doxycycline, although activity of ϳ1 ϫ 10 9 cpm/mg. The hybridization cocktail contained 50% some expression is seen in dynorphin neurons as well. Controls and formamide, 4ϫ SSC (1ϫ SSC is 0.15 M NaCl and 0.015 sodium citrate, ⌬FosB-overexpressing mice are littermates within each line (11A and pH 7.0), 1ϫ Denhardt’s solution, 1% sarcosyl, 0.02 M Na PO , pH 7.0, 11B) and have the same bitransgenic construct, which can be activated by 3 4 10% dextran sulfate, 0.06 M dithiothreitol, and 0.1 mg/ml sheared salmon removal of doxycycline. All mice were conceived and raised on the sperm DNA. Hybridization was performed for 18 hr in a humidified tetracycline derivative doxycycline at a dose of 100 g/ml in the drinking chamber at 42°C. After hybridization, the sections were rinsed four times water. As adults, one-half of the resulting litters were maintained on for 20 min each in 1ϫ SSC at 60°C. Thereafter, the sections were rinsed doxycycline (controls); the other half were removed from doxycycline in autoclaved water for 10 sec, dehydrated in alcohol, and air-dried. (⌬FosB overexpressers) for the rest of the experiment. Six weeks after Finally, NTB2 nuclear track emulsion (diluted 1:1 with water; Kodak, removal of doxycycline, at which time ⌬FosB expression is known to be Rochester, NY) was applied by dipping. After 2–4 weeks of exposure, maximal (Chen et al., 1998; Kelz et al., 1999), the running wheels were the slides were developed with D19 (Kodak) and fixed with Unifix unlocked for both mice on tetracycline (controls) and mice on tap water (Kodak).