European Journal of Pharmacology 398 Ž2000. 259–262 www.elsevier.nlrlocaterejphar Short communication Differential effects of ibogaine on behavioural and dopamine sensitization to cocaine Karen K. Szumlinski ) , Isabelle M. Maisonneuve, Stanley D. Glick Center for Neuropharmacology and Neuroscience, MC-136, Albany Medical College, 47 New Scotland AÕenue, Albany, NY 12208, USA Received 2 March 2000; received in revised form 14 April 2000; accepted 20 April 2000 Abstract To investigate a possible basis for the proposed anti-addictive property of ibogaine, the effects of ibogaine Ž40 mgrkg, i.p., 19 h earlier. on the expression of sensitization induced by cocaine were investigated. Ibogaine pretreatment potentiated the increase in the stereotypic effects of a cocaine challenge Ž20 mgrkg. in both sensitized Ž5 = 15 mgrkg, i.p.. and acutely treated rats. However, while ibogaine pretreatment did not significantly alter the dopamine response in the nucleus accumbens to acute cocaine, it abolished the expression of cocaine-induced dopamine sensitization. This result demonstrates that ibogaine pretreatment can reverse one of the neuroadaptations produced by chronic cocaine administration, an effect that may contribute to its putative anti-addictive property. q 2000 Elsevier Science B.V. All rights reserved. Keywords: Ibogaine; Cocaine; Sensitization; Dopamine; Stereotypy; Drug addiction 1. Introduction The naturally occurring indole alkaloid, ibogaine, is being investigated currently for its putative ‘‘anti-addictive’’ properties Žfor review, see Glick and Maisonneuve, 1998.. Although the neural mechanismŽs. underlying ibogaine’s effects are still unclear, drug experience appears to render an animal more sensitive to the effects of ibogaine and related agents on drug-induced behaviour Žfor review, see Glick et al., 2000; Szumlinski et al., 2000c.. For example, pretreatment Ž19 h earlier. with either ibogaine or a synthetic congener, 18-methoxycoronaridine Ž18-MC., can produce a greater increase in the motor effects of both cocaine ŽSzumlinski et al., 1999a,b,c, 2000d. and methamphetamine ŽSzumlinski et al., 2000a. in stimulant-experienced vs. stimulant-naive rats. Repeated, intermittent stimulant exposure can induce a sensitization of dopamine transmission in the mesolimbic pathway and this action is implicated in both the motorsensitizing Že.g., Kalivas and Stewart, 1991. and addictive effects Že.g., Kalivas et al., 1993; Robinson and Berridge, ) Corresponding author. Tel.: q1-518-262-5801; fax: q1-518-2625799. E-mail address: szumlik@mail.amc.edu ŽK.K. Szumlinski.. 1993, but see Di Chiara, 1999. of many drugs of abuse. A corollary of the sensitization theories of addiction is that anti-addictive drugs should block the expression of sensitization Žbe it behavioural andror neurochemical.. The present study examined this hypothesis using the putative anti-addictive drug, ibogaine Že.g., Frenken, 2000; Sheppard, 1994., in rats with previous cocaine experience. 2. Materials and methods 2.1. Drugs Ibogaine HCl and cocaine HCl, obtained from Sigma, were dissolved in water and saline, respectively, and were injected i.p. in doses expressed as the salt. 2.2. Design and procedures Female Sprague–Dawley rats were implanted stereotaxically with a guide cannula over the shell nucleus accumbens, as described previously ŽSzumlinski et al., 2000b.. The day after surgery, rats were randomly assigned to groups that received once daily injections of either saline or cocaine Ž15 mgrkg. for 5 days. Following the 5th injection, animals were withdrawn from chronic 0014-2999r00r$ - see front matter q 2000 Elsevier Science B.V. All rights reserved. PII: S 0 0 1 4 - 2 9 9 9 Ž 0 0 . 0 0 3 2 5 - 3 260 K.K. Szumlinski et al.r European Journal of Pharmacology 398 (2000) 259–262 treatment for 2 weeks. On the last day of withdrawal, rats were placed in a dialysis chamber where a calibrated probe Ž2 mm probe, CMA. was lowered into the guide cannula. The dialysis probe was continuously perfused with artificial cerebral spinal fluid Ž146 mM NaCl, 2.7 mM KCl, 1.2 mM CaCl 2 and 1.0 mM MgCl 2 . at a rate of 1 mlrmin. Animals were then pretreated with either ibogaine Ž40 mgrkg, i.p.. or vehicle. The next day Ž1100 h., collection of perfusates began. After 2 h of baseline collections, the rats received their test injection of cocaine Ž20 mgrkg, i.p.. Ž1300 h, 19 h following ibogaine pretreatment.. The collection of dialysate samples was then continued for 3 h. Table 1 Effects of ibogaine Ž40 mgrkg, 19 h earlier. or vehicle on the basal levels of dopamine and its metabolites, DOPAC and HVA of rats treated chronically with either cocaine Ž5=15 mgrkg. or saline. Data are expressed as pmolr10 ml Žmean"S.E.M.. Chronic saline Vehicle Ž ns8. Chronic cocaine Ibogaine Ž ns6. Vehicle Ž ns 7. Ibogaine Ž ns 7. Dopamine 0.007"0.000 0.008"0.002 0.005"0.001 0.007"0.002 DOPAC 5.290"1.333 9.330"1.333 7.801"1.765 3.942"1.432 ) HVA 2.945"0.637 1.849"0.085 3.467"0.0526 1.814"0.582 ) ) Denotes P - 0.05 vs. respective Chronic saline group ŽDuncan’s Multiple Range post hoc tests.. 2.3. Catecholamine assay Dialysate samples from brains in which the probe was accurately placed within "0.5 mm of the shell of the nucleus accumbens were assayed for dopamine, 3,4-dihydroxyphenylacetic acid ŽDOPAC., and homovanillic acid ŽHVA. by high-performance liquid chromatography ŽHPLC. with electrochemical detection using an HPLC system with an ESA Coulochemm II electrochemical detector, as described previously ŽSzumlinski et al., 2000b.. line sampling. Compared to vehicle animals, ibogaine pretreatment lowered the basal concentrations of both DOPAC and HVA in chronic cocaine rats only wfor DOPAC: Chronic Treatment= Pretreatment, F Ž1,24. s 6.02, P s 0.022; for HVA: interaction, F Ž1,24. s 5.71, P s 0.03x Žsee Table 1.. 2.4. BehaÕioural scoring A robust sensitization of extracellular levels of dopamine was observed in response to the challenge injection of cocaine in vehicle-pretreated chronic cocaine rats, compared to vehicle-pretreated acute controls wChronic Treatment = Time, F Ž14,182. s 4.93, P - 0.0001x ŽFig. 1, top.. No sensitization of either DOPAC Žinteraction, P s 0.95. or HVA Žinteraction, P s 1.0. was observed Ždata not shown.. Ibogaine produced differential effects on cocaine-induced increases in accumbal dopamine in acute vs. chronic cocaine treated rats wChronic Treatment= Pretreatment= Time, F Ž14, 336. s 3.32, P - 0.0001x; compared to vehicle controls, ibogaine did not affect dopamine levels in acute cocaine rats ŽPretreatment= Time interaction, P s 0.80., but lowered dopamine levels in cocaine sensitized rats winteraction, F Ž14,168. s 3.38, P - 0.0001x ŽFig. 1, top.. Overall, ibogaine pretreatment lowered the extracellular levels of DOPAC wPretreatment= Time, F Ž14,336. s 2.02, P s 0.02x and HVA winteraction, F Ž14,336. s 1.94, P s 0.02x in response to cocaine Ždata not shown.. A behavioural intensity rating scale, adapted from Kalivas et al. Ž1988., was used to quantify the stereotypy expressed by the animals during the injection sessions. Rats were observed for 30 s, during the minute that preceded dialysate removal Ži.e., every 20 min., beginning at the last hour of baseline dialysate sampling and continuing until the completion of the microdialysis session. Behaviour was rated as follows: Ž1. asleep or still; Ž2. inactive, grooming or mild licking; Ž3. locomotion Žall four feet move in 30 s., rearing or sniffing; Ž4. any combination of two of locomotion, rearing or sniffing; Ž5. continuous sniffing for 30 s without locomotion or rearing; Ž6. continuous sniffing for 30 s with locomotion or rearing; Ž7. patterned sniffing for 15 s; Ž8. patterned sniffing for 30 s; Ž9. continuous gnawing or focused grooming Ž‘‘skin-picking’’-like behaviour.; and Ž10. bizarre diskinetic movements or seizures. 3. Results 3.1. Baseline catecholamine concentrations Neither chronic cocaine treatment Ž5 = 15 mgrkg. ŽChronic effect, P s 0.34. nor ibogaine pretreatment Ž40 mgrkg. ŽPretreatment effect, P s 0.39. altered the basal concentrations of dopamine in the accumbens during base- 3.2. Test for dopamine sensitization 3.3. Test for behaÕioural sensitization As evidenced in Fig. 1 Žbottom., a sensitization of stereotypy was observed in response to the challenge injection of cocaine in vehicle-pretreated chronic cocaine rats, compared to vehicle-pretreated chronic saline controls wChronic Treatment = Time, F Ž12,168. s 3.07, P 0.001x. Ibogaine potentiated the stereotypic activity of both chronic saline and chronic cocaine treated groups wPretreat- K.K. Szumlinski et al.r European Journal of Pharmacology 398 (2000) 259–262 261 Fig. 1. Effects of ibogaine Ž40 mgrkg, 19 h earlier; solid. or vehicle Žopen. on the timecourses of extracellular levels in the accumbens of dopamine Žtop. and the stereotypic behaviour Žbottom. of rats treated chronically with either cocaine Ž5 = 15 mgrkg; right. or saline Žleft. in response to a challenge injection of cocaine Ž20 mgrkg.. Each data point represents the mean percent of baseline of six to eight rats at the indicated times during testing. S.E.M.s ranged from 2% to 30% of the mean for dopamine and from 0% to 2% of the mean for stereotypy. ) P - 0.05 vs. chronic saline; qP - 0.05 vs. vehicle ŽDuncan Multiple Range post-hoc tests.. ment = Time, F Ž12, 312. s 3.14, P - 0.00; no Chronic Treatment= Pretreatment= Time interaction, P s 0.25x. 4. Discussion As the phenomenon of sensitization has been theoretically implicated in drug addiction, it was hypothesized that the putative ‘‘anti-addictive’’ drug, ibogaine, should block sensitization to cocaine. Consistent with this hypothesis and the results of previous studies ŽSzumlinski et al., 2000b,d., ibogaine pretreatment Ž40 mgrkg, 19 h earlier. abolished the expression of dopamine sensitization in the nucleus accumbens in rats with previous cocaine experience Ž5 = 15 mgrkg.. The present results provide further evidence for a relationship between the attenuating effects of iboga agents on drug self-administration in rodents and their ability to block the expression of dopamine sensitization produced by the repeated administration of drugs of abuse. Recent reports demonstrated that the expression of behavioural sensitization to morphine ŽCadoni and Di Chiara, 1999., nicotine ŽCadoni and Di Chiara, 2000., amphetamine and cocaine ŽCadoni et al., 2000. are coincident with a sensitization of dopamine levels in the nucleus accumbens core. In the cases of cocaine and nicotine, a reduction in dopamine levels in the accumbens shell has been observed in relation to the expression of behavioural sensitization ŽCadoni and Di Chiara, 2000; Cadoni et al., 2000.. Consistent with this, at least in ibogaine pretreated rats, the magnitude of the behavioural response to cocaine did not correspond with that of the dopamine response in the nucleus accumbens shell. This discrepancy was most notable in the chronic cocaine group where ibogaine pretreatment exerted opposite effects on the expression of motor sensitization Žpotentiation. and dopamine sensitization in the accumbens shell Žblockade.. Thus, ibogaine might exert its potentiating effects on motor behavioural sensitization by blocking the increase in dopamine responsiveness in the shell that serves normally to counteract the sensitization of core dopamine responsiveness. However, a number of past ŽGlick et al., 2000; Szumlinski et al., 2000b,d. and present results contrast directly with those of Cadoni et al. Ž1999, 2000.. First, in our hands, the chronic administration of drugs of abuse produces a sensitization of both dopamine responsiveness in the nucleus accumbens shell ŽGlick et al., 2000; Szumlinski et al., 2000b; present study. and motor behaviour Žfor review, see Szumlinski et al., 2000c.. Moreover, in more than one study, the time-courses of both the acute and sensitized motor responses to cocaine are coincident with the changes in extracellular levels of dopamine as measured in the accumbens shell ŽGlick et al., 2000; Szumlinski et al., 2000d; present study.. Lastly, no reduction in the 262 K.K. Szumlinski et al.r European Journal of Pharmacology 398 (2000) 259–262 shell dopamine response to cocaine was observed in any chronic drug treated groups tested to date ŽSzumlinski et al., 2000d; present study.. From the available data, we hypothesize that the effects of ibogaine and related agents on drug-induced motor sensitization and on drug self-administration are mediated by different underlying mechanisms. The anatomical connectivity Že.g., Jongen-Relo et al., 1993. and function ŽIkemoto and Panksepp, 1999; Sokolowski and Salamone, 1998. of the core and shell regions of the nucleus accumbens are distinct. 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