Neuropsychopharmacology (2009) 34, 1946–1957 & 2009 Nature Publishing Group All rights reserved 0893-133X/09 $32.00 www.neuropsychopharmacology.org Role of Mu- and Delta-Opioid Receptors in the Nucleus Accumbens in Cocaine-Seeking Behavior 1 ,1 Diana Simmons and David W Self* 1Department of Psychiatry and the Neuroscience Graduate Program, The Seay Center for Basic and Applied Research in Psychiatric Illness, UT Southwestern Medical Center, Dallas, Texas, USA Earlier studies suggest that opioid receptors in the ventral tegmental area, but not the nucleus accumbens (NAc), play a role in relapse to drug-seeking behavior. However, environmental stimuli that elicit relapse also release the endogenous opioid b-endorphin in the NAc. Using a within-session extinction/reinstatement paradigm in rats that self-administer cocaine, we found that NAc infusions of the mu- opioid receptor (MOR) agonist DAMGO moderately reinstated responding on the cocaine-paired lever at low doses (1.0–3.0 ng/side), whereas the delta-opioid receptor (DOR) agonist DPDPE induced greater responding at higher doses (300–3000 ng/side) that also enhanced inactive lever responding. Using doses of either agonist that induced responding on only the cocaine-paired lever, we found that DAMGO-induced responding was blocked selectively by pretreatment with the MOR antagonist, CTAP, whereas DPDPE-induced responding was selectively blocked by the DOR antagonist, naltrindole. Cocaine-primed reinstatement was blocked by intra-NAc CTAP but not naltrindole, indicating a role for endogenous MOR-acting peptides in cocaine-induced reinstatement of cocaine-seeking behavior. In this regard, intra-NAc infusions of b-endorphin (100–1000 ng/side) induced marked cocaine-seeking behavior, an effect blocked by intra-NAc pretreatment with the MOR but not DOR antagonist. Conversely, cocaine seeking elicited by the enkephalinase inhibitor m thiorphan (1–10 g/side) was blocked by naltrindole but not CTAP. MOR stimulation in more dorsal caudate-putamen sites was ineffective, whereas DPDPE infusions induced cocaine seeking. Together, these findings establish distinct roles for MOR and DOR in cocaine relapse and suggest that NAc MOR could be an important therapeutic target to neutralize the effects of endogenous b- endorphin release on cocaine relapse. Neuropsychopharmacology (2009) 34, 1946–1957; doi:10.1038/npp.2009.28; published online 11 March 2009 Keywords: reinstatement; b-endorphin; thiorphan; DAMGO; DPDPE; relapse INTRODUCTION cocaine seeking elicited by exposure to cocaine-associated cues (Burattini et al, 2008). Similarly, cocaine-primed Drug addiction involves dysregulation in brain reward cocaine seeking is blocked by systemic administration of circuitry leading to compulsive drug use (Dackis and the partial mu-opioid receptor (MOR) agonist buprenor- O’Brien, 2001; Kalivas and Volkow, 2005; Koob and Le phine, the delta-opioid receptor (DOR) antagonist naltrin- Moal, 2001). In addition to drug reward, the mesolimbic dole, or the nonspecific opioid antagonist naltrexone dopamine system plays an integral role in relapse to drug- (Comer et al, 1993; Gerrits et al, 2005). seeking behavior, as stimuli that elicit drug seeking also Stewart (1984) and colleagues found that opioid receptors activate dopamine neurons in the ventral tegmental area in the VTA play a role in reinstatement of cocaine and (VTA) leading to dopamine release in forebrain regions, heroin seeking, as intra-VTA morphine treatments trigger such as the nucleus accumbens (NAc) (Phillips et al, 2003; drug seeking in an extinction/reinstatement paradigm, an Pruessner et al, 2004; Self and Nestler, 1998; Shalev et al, animal model of relapse. This effect is thought to involve 2002; Spealman et al, 1999; Stewart, 2000). Opioid receptors local disinhibition of dopamine neurons in the VTA, also play a role in relapse to cocaine seeking in animal leading to dopamine release in the NAc (Ford et al, 2006; models, as systemic treatment with naltrexone inhibits Johnson and North, 1992; Leone et al, 1991). In contrast, earlier studies suggest that opioid receptors localized in the NAc do not play a role in drug seeking, as intra-NAc *Correspondence: Dr DW Self, Department of Psychiatry, University of Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX morphine treatments fail to reinstate cocaine or heroin 75390-9070, USA, Tel: + 214 648 1237, Fax: + 214 648 4947, seeking (Stewart and Vezina, 1988; Tang et al, 2005), and E-mail: [email protected] blockade of NAc MOR with the selective MOR antagonist Received 8 November 2008; revised 28 January 2009; accepted 5 CTAP (D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2)failed February 2009 to significantly alter cocaine-primed reinstatement of Mu and Delta Agonists Trigger Cocaine Relapse D Simmons and DW Self 1947 cocaine seeking (Tang et al, 2005). However, endogenous Sucrose Lever Press Training and Surgery opioid peptides, such as b-endorphin, are released in the Lever-press training, self-administration, and reinstatement NAc by cocaine, and stressful situations (Roth-Deri et al, testing were performed in operant test chambers (Med- 2004, 2003; Zangen and Shalev, 2003), events that trigger the Associates, East Fairfield, VT). Chambers were equipped reinstatement of cocaine seeking, and intra-NAc morphine with two response levers and an infusion pump as described infusions induce a conditioned place preference (van der Kooy et al, 1982). earlier (Edwards et al, 2007). Animals were food-restricted to prevent weight gain and trained to lever-press for sucrose Opioid receptors are highly expressed by NAc neurons pellets on a fixed ratio 1 (FR1) reinforcement schedule until (Mansour et al, 1995, 1987), and local opioid infusions in an acquisition criteria of 100 sucrose pellets consumed for 3 the NAc modulate behavior in a biphasic manner. Thus, 2 4 5 consecutive test days was reached. Following lever-press microgram doses of (D-Ala ,N-Me-Phe ,glycinol )-enkepha- training, animals were fed ad libitum for at least 1 day lin (DAMGO) (MOR agonist) or morphine infused in the before surgery. Animals were anesthetized and implanted NAc initially suppress locomotion but subsequently induce with a chronic indwelling catheter into the jugular vein that hyper locomotion (Cunningham and Kelley, 1992; Meyer exited subcutaneously on the back. An intracranial, 26- et al, 1994). Lower doses of DAMGO decrease the latency for guage bilateral guide cannula was aimed at the NAc hyper locomotion to occur (Meyer et al, 1994). As doses of ± À agonists used in these locomotor studies are similar to those ( 1.5 mm lateral; 1.7 mm anterior to bregma; 5.7 ventral to dura with the level skull) or caudate putamen (CPu) used in prior reinstatement studies, it is possible that the (±1.5 mm lateral; 1.7 mm anterior to bregma; À3.2 mm behavioral suppressive effects masked the potential of NAc opioid receptor stimulation to trigger the reinstatement of ventral to dura) (Paxinos and Watson, 1998). Dummy and infusion cannulae (33 gauge) were cut to extend 1 mm drug-seeking behavior. Moreover, NAc infusions of opioid beyond the guide cannulae tip, and dummy cannulae agonists induce feeding behavior but also with a prolonged remained in place until the day of intracranial drug latency to initiate feeding (Bakshi and Kelley, 1993; Kelley infusion. Animals were allowed 5–7 days to recover before et al, 2005). Similarly, intra-NAc infusions of DAMGO starting the experiment. increase the motivation for food on a progressive ratio reinforcement schedule, and where response breakpoints are obtained after some delay (Zhang et al, 2003). Therefore, Cocaine Self-Administration and Within-Session it is possible that these delayed motivational effects reflect Reinstatement Testing the initial suppressive effects of high-dose MOR agonist infusions. Animals were tested in a within-session extinction/rein- In this study, we investigated the role of NAc opioid statement paradigm as described earlier (Bachtell et al, receptors in the reinstatement of cocaine-seeking 2005). Briefly, animals self-administered cocaine (0.5 mg/kg behavior using MOR- and DOR-selective ligands and in 0.1 ml over 5 s, time-out period 15 s) in daily 4 h sessions endogenous opioid peptides. We found that NAc infusions for 5–6 days/week until a criteria of 3 consecutive days of of MOR and DOR agonists effectively reinstate cocaine o10% variance in mean cocaine intake was reached (B3 seeking through selective actions at their respective weeks). During the 5s injection, a cue light above the lever receptors. Stimulation of NAc opioid receptors by the was illuminated, whereas the house light was turned off for endogenous peptides, b-endorphin and enkephalins, also the entire injection and time-out period. Subsequently, induced cocaine-seeking behavior. The results clearly animals were trained in the within-session extinction establish that either MOR or DOR stimulation in the NAc paradigm that consisted of 1 h cocaine availability followed is sufficient to elicit cocaine-seeking behavior, and that by 3 h extinction conditions in which only response- MOR receptors play an important role in cocaine-primed contingent injection cues were available. Animals extin- relapse. These findings also suggest that persistent neuroa- guished responding to criteria of p5 responses at either the daptations in NAc opioid receptors following chronic drug-paired or the inactive lever for the final hour of the cocaine use could contribute to drug-seeking behavior in session for at least three consecutive sessions, while prolonged abstinence. maintaining a minimum of 15 self-administered cocaine injections with ±10% variability in the first hour of the session. Mean cocaine self-administration on the test day was 25±0.74 (NAc) and 26±0.95 (CPu) injections/h. Test days were conducted with an intra-NAc infusion of MOR MATERIALS AND METHODS and DOR agonists alone (0.5 ml/side over 2 min) or in Animals and Housing Conditions combination as sequential antagonist/agonist infusions (1.0 ml/side total volume) immediately before the final hour Male Sprague–Daley rats weighing 225–275 g (Charles River of the test session.