sign in sign up
<<
Home , Deliriant, Dizocilpine, Hallucinogen, Rapastinel, Salvinorin A

ZEBRAFISH Volume 13, Number 5, 2016 Review Articles ª Mary Ann Liebert, Inc. DOI: 10.1089/zeb.2016.1251

Exploring and Psychedelic Medicine with Zebrafish Models

Evan J. Kyzar1 and Allan V. Kalueff2–6

Abstract

After decades of sociopolitical obstacles, the field of is experiencing a revived interest in the use of hallucinogenic agents to treat disorders. Along with the use of for , recent pilot studies have highlighted the efficacy of classic , such as lysergic acid diethylamide and , in treating , post-traumatic stress disorder, and . However, many basic phar- macological and toxicological questions remain unanswered with regard to these compounds. In this study, we discuss psychedelic medicine as well as the behavioral and toxicological effects of hallucinogenic in zebrafish. We emphasize this aquatic organism as a model ideally suited to assess both the potential toxic and therapeutic effects of major known classes of hallucinogenic compounds. In addition, novel drugs with hal- lucinogenic properties can be efficiently screened using zebrafish models. Well-designed preclinical studies utilizing zebrafish can contribute to the reemerging treatment paradigm of psychedelic medicine, leading to new avenues of clinical exploration for psychiatric disorders.

Introduction requiring the use of both traditional (rodent) and novel model species.6 In this study, we briefly review the history of psy- allucinogenic drugs have been used by humans for chedelic medicine and the pharmacology of these drugs and centuries, exerting potent effects on , cognition, H discuss the effects of hallucinogenic drugs on larval and adult and behavior with little propensity for habit formation and zebrafish (Danio rerio). We highlight the utility of this aquatic addiction.1 In 1940s and 1950s, scientists began to realize the model organism in future experiments investigating the po- potential of these compounds in neuroscience and psychiatry. tential behavioral benefits and pharmacological toxicity of However, laws have categorized many hallucinogens in hallucinogenic compounds. the most strict regulatory groupings (e.g., Schedule I in the United States and Class A in the United Kingdom), leading to Psychedelic Medicine and Pharmacology a relative dearth of scientific inquiry into the biology of these compounds.2,3 Nonetheless, the last decade has seen a revival While the term hallucinogen often refers to different classes of hallucinogenic drug investigation,4,5 with a particular focus of drugs with -altering properties, this definition may be on identifying compounds that may have efficacy in treating overly broad and reliant on terminology that principally in- intractable psychiatric illnesses. volves visual phenomenology.7 Earlier studies have limited While the use of hallucinogens in medicine shows promise, this term to a more narrow definition, only ascribing to the these compounds are not without risk. The years of scientific word to classic serotonergic hallucinogens, such as lysergic dormancy caused, in part, by governmental regulation and acid diethylamide (LSD) and , with proper- 7,8 societal taboo have left many unanswered questions regarding ties at the 5HT2A . However, here, we the pharmacology and toxicity of hallucinogens in vivo.Thus, classify hallucinogens into three broader categories: (1) classic high-throughput screening techniques are needed to test for (serotonergic) psychedelics, (2) , and (3) drug interactions and aberrant behavioral effects if hallucino- (Fig. 1). Classic serotonergic hallucinogens bind to serotonin gens are to gain traction as potential medicinal options, receptors and generally exhibit agonist properties. Previous

1Department of Psychiatry, College of Medicine, University of Illinois at Chicago, Chicago, Illinois. 2Research Institute for Marine Drugs and Nutrition, College of Food and Technology, Guangdong Ocean University (GDOU), Zhanjiang, China. 3ZENEREI Institute, Slidell, Louisiana. 4Institute of Translational Biomedicine, St. Petersburg State University, St. Petersburg, Russia. 5Institutes of Chemical Technology and Natural , Ural Federal University, Ekaterinburg, Russia. 6The International Zebrafish Neuroscience Research Consortium (ZNRC), Slidell, Louisiana.

379 380 KYZAR AND KALUEFF

FIG. 1. Summary of principal pharmacological and behavioral effects of selected classes of hallucinogenic drugs. ‘‘Mixed’’ drugs that have high affinity for more than receptor shown here (e.g., ) are not included in the present chart. Note that all drugs listed possess some binding affinity for other receptors. LSD, lysergic acid diethylamide; PCP, . studies have suggested that agonism at the 5HT2A receptor, as nogen combining pharmacological properties of several well as substitution for the prototypical hallucinogen 2,5- classes of psychoactive drugs.18 dimethoxy-4-methylamphetamine (DOM) in discrimination Humans have long been using hallucinogenic drugs for tasks, be a requisite for inclusion in this category.8,9 Classic spiritual and religious purposes, utilizing natural and serotonergic hallucinogens can be further divided into two products that produce psychoactive effects when ingested.19 structural classes— indoleamines (including LSD and N,N- For example, an preparation consisting of Banis- dimethyltryptamine [DMT]) and phenylalkylamines (includ- teriopsis caapi vines brewed with leaves from members of the ing mescaline, DOM, and the ‘‘’’ class of drugs, such as 2,5- genus Psychotria is consumed by tribal people of the Amazon dimethoxy-4-bromophenethylamine [2C-B]). Basin in a ritualistic and sacramental context.20,21 Arguably, Clinically, hallucinogens produce a profound the Western conceptualization of hallucinogenic drugs began state of disconnection from reality and the surrounding en- in the late 1800s, when Arthur Heffter isolated mescaline from vironment.10 Ketamine and phencyclidine (PCP), the most the (Lophophora williamsii) and confirmed its commonly recognized drugs in this category, produce their subjective effects through self-experimentation.22 This was effects through antagonism at the N-methyl-D-aspartate followed by the chance discovery of LSD by (NMDA) receptor.11 Although acting through j- in 1943,23 and his subsequent isolation of psilocybin from agonism rather than NMDA receptors, (the ac- ‘‘ ’’ in 1958.24 tive hallucinogenic component of the divinorum) These early studies coalesced with growing countercul- is also considered a dissociative hallucinogen.12 tural influences, leading to a period of rapid growth in both The third group is defined by -inducing agents scientific and social testing of hallucinogenic compounds. Of principally composed of derived from plants, such particular interest was the use of LSD in , gi- as belladonna (deadly nightshade) and stra- ven the ability of this drug to evoke experiences described as monium (Jimson weed), which act as ‘‘otherworldly’’ that may confer long-lasting changes to pa- agents.13,14 Typical drugs, including and tients’ behavior.25 Beginning in the 1950s, numerous ex- , cause vivid along with physical periments probed the benefits of LSD-assisted psychotherapy symptoms of anticholinergic toxicity (e.g., dry mouth, dia- in disease states ranging from to terminal cancer phoresis, , and death) that are usually unpleasant diagnoses.26–29 Other studies investigated the potential cog- in nature, leaving many recreational users unwilling to repeat nitive enhancement effects of psychedelic drugs, such as the the experience. Some drugs that are considered hallucino- ability to improve and problem solving.30 genic do not comfortably fit in one of the above categories. Although some early studies utilizing LSD and other hal- For example, the drug ibogaine is a naturally occurring al- lucinogenic drugs resulted in positive clinical outcomes, re- kaloid that can be isolated from members of the Apocynaceae search into psychedelic medicine halted in the early 1970s family of plants. Ibogaine possesses binding affinity for both with the passage of the United States Controlled Substances 5HT2A and NMDA receptors, as well as activity at other Act and the United Kingdom Misuse of Drugs Act. Nearly all serotonergic, , , and sigma recep- hallucinogenic drugs were placed in Schedule I (United tors,15–17 therefore representing a ‘‘mixed-action’’ halluci- States) or Class A (United Kingdom), implying that these PSYCHEDELIC MEDICINE AND ZEBRAFISH 381 agents have no recognized medical use. This restriction led to creased anxiety-like behavior) along with whole-body cor- a period of relatively little scientific inquiry into the biology tisol and brain c-fos expression (serving as a marker of and potential therapeutic effects of hallucinogens. Despite neuronal activation).51 The dissociation between observed this period of stagnation, a revival of scientific inquiry into behavioral anxiolysis and cortisol levels is interesting and the beneficial effects of hallucinogenic drugs has begun5,31 in may be related to increased serotonergic tone and subse- both the basic6,8 and clinical laboratories.32 The striking quent activation of the stress axis.51 LSD also decreased finding that ketamine rapidly alleviates depressive symptoms shoaling behavior in adult zebrafish.52 Mescaline treatment has further triggered a reevaluation of this and other similar led to a similar increase in top dwelling in the NTT but in- drugs in the psychiatric setting.33–36 However, the antide- creased adult zebrafish shoaling behavior and did not change pressant effects of ketamine fade quickly (approximately 1–2 whole-body cortisol levels53 (Table 1). Interestingly, psilocy- weeks after infusion), and other nonhallucinogenic drugs, bin had no observable effects on zebrafish behavior but sig- D- and (that also act at the NMDA nificantly increased whole-body cortisol.54 receptor), are also being investigated for their possible ther- Although not normally considered a classic serotonergic apeutic effects.37 hallucinogen, MDMA also decreased anxiety-like behaviors Classic serotonergic hallucinogens have gained traction as and shoaling in adult zebrafish,52,55 acting similar to LSD. novel treatments for psychiatric disorders. For example, a Interestingly, the mixed serotonergic/dissociative hallucinogen 2012 meta-analysis of six studies of LSD in addiction ibogaine tended to reverse innate phenotypes of adult zebrafish revealed a significant treatment effect,38 as LSD-treated pa- in the NTT and the light–dark box (LDB) models. For exam- tients improved at the first follow-up compared to control ple, adult zebrafish usually begin the NTT by exploring the patients in all six included studies. Recently, alcoholic pa- bottom of the tank and gradually entering and exploring the top tients showed a lasting decrease in drinking after psilocybin half of the tank.56 In contrast, fish exposed to ibogaine show an administration, and, interestingly, the intensity of subjective unusual ‘‘inverted’’ pattern of habituation, exploring the top drug effects correlated with the magnitude of decrease in half of the tank first in the NTT and exploring the light com- alcohol intake.39 Likewise, 80% of -dependent pa- partment before the dark compartment in the LDB.57 It is tients who underwent a 15-week cognitive behavioral therapy possible that the ability of ibogaine to affect zebrafish behavior regimen combined with two to three sessions of psilocybin may be related to its well-studied ability to reduce craving and exposure remained abstinent from 36 weeks after drug use in -dependent patients.58 While the exact treatment, which exceeds the *35% success rates at this mechanism of ibogaine’s effect on addiction remains un- same time point for other common behavioral and pharma- known, the inversion of behavioral patterns in zebrafish may be cological interventions.40 reflective of an ability to reverse longstanding behaviors in Classic serotonergic hallucinogens have been investigated addicted patients as well. Additionally, the interplay between in other psychiatric disorders, including post-traumatic stress anxiety and addiction is well studied, and ibogaine (as well as disorder (PTSD) and anxiety associated with terminal can- its principal metabolite, ) reduced anxiety-like cer.41 A preliminary trial suggested that LSD combined with measures in the NTT and LDB.57,59 To the best of our psychotherapy may potentially reduce anxiety-related out- knowledge, no larval zebrafish studies have investigated the comes in patients with end-stage illnesses, and follow-up effects of classic serotonergic hallucinogens to date. revealed that the LSD-induced alleviation of symptoms las- ted for at least a year after treatment.42,43 A similar study Dissociative hallucinogens investigated psilocybin in patients with end-of-life-associated anxiety, finding decreased anxiety and increased measures of Dissociative hallucinogens are perhaps the best-studied positive affect in drug-treated patients.44 Notably, these studies class of psychedelic drugs in zebrafish. For example, ketamine did not reveal any clinically significant deleterious effects of is both neurotoxic and teratogenic in developing zebrafish, LSD or psilocybin. 3,4-Methylenedioxymethamphetamine but acetyl-L-carnitine supplementation may reverse or res- (MDMA, Ecstasy) also has beneficial effects in patients suf- cue some ketamine-induced neurodevelopmental abnormali- fering from PTSD in multiple clinical trials showing persistent ties.60–64 Ketamine also reduces cytochrome p450 aromatase positive outcomes.45–47 It should be noted that MDMA is often levels and estradiol expression in larval fish65 and exerts an- not considered a classic serotonergic hallucinogen, although it xiolytic effects on adult zebrafish in multiple behavioral acts through serotonergic and mechanisms and tests.66–68 It also evokes robust circling behavior in zebrafish— overlaps with some clinical effects of psychedelics.7,48–50 an effect seen following antagonism in rodent models66,67,69,70 and likely related to dissociative-like effects in Zebrafish Models of Hallucinogenic Drug Exposure humans.71 Consistent with this, PCP, another NMDA antago- nist with dissociative effects, also caused effects and The revival in laboratory testing of hallucinogenic drugs stereotypic circling behavior in adult zebrafish.53 has spread to include testing these drugs in aquatic models, The potent NMDA antagonist MK-801 exerts effects on such as zebrafish.18 Here, we summarize some of the most neuronal development in larval zebrafish, particularly affecting recent data and tests of hallucinogens using zebrafish models early retinotectal visual projections,72–74 increasing locomo- (see Table 1 for details). tor activity in a dose/strain manner75,76 and impairing learn- ing and memory in a visual lateralization test.77 Similar to Classic serotonergic hallucinogens ketamine, MK-801 induces circling behavior in adult zebra- LSD elicits robust anxiolytic effects in adult zebrafish fish, although this may be a consequence of MK-801-induced in the novel tank test (NTT), characteristically increasing hyperlocomotion78–80 peaking in adulthood.81 MK-801 also top-dwelling behavior (typically considered a marker of de- interferes with the acquisition of learning and memory tasks in Table 1. Summary of Known Hallucinogenic Drug Effects in Zebrafish Reward- Shoaling Circling related Cortisol Selected Drug class Drug Locomotion Anxiety behavior behavior behavior levels gene expression Larval studies References 51,52 Classic LSD NC Decreased Decreased Noa N/A Increased Increased brain c-fos N/A 53,92 serotonergic Mescaline NC Decreased Increased Noa N/A NC N/A N/A hallucinogens a 54 Psilocybin NCa NCa Increased/NC Noa N/A Increased N/A N/A 57 Mixed Ibogaine Increased Decreased Decreased No N/A NC NC (brain N/A c-fos) 63,66,67 Dissociatives Ketamine NC Decreased/ Decreased Yes N/A Decreased Increased brain c-fos; Neurotoxic, Increased Decreased phox2b teratogenic 382 (ketamine alone) and sirt1 (ketamine + ) 73,75,78,80,86 MK-801 Increased Decreased/NC Decreased Yes N/A N/A N/A Altered retinotectal patterning 53,54 PCP NC Decreased Decreased Yes N/A Increased N/A N/A 91,92 Salvinorin A Increased Decreased/NCa NCa N/A Increased NCa NCa (brain c-fos) N/A CPP Deliriants Atropine N/A N/A N/A N/A N/A N/A N/A Neurotoxic 173,174 94,96 Scopolamine NC NC N/A N/A N/A N/A N/A N/A

aOwn unpublished observations. Note a general reduction of anxiety by various hallucinogenic drugs in zebrafish, illustrating the potential of this aquatic animal model for studying anxiolytic therapeutic potential of hallucinogens. The colors used represent the following: dark gray = increased, medium gray = decreased, other = conflicting reports. White color denotes not assessed (N/A) phenotypes, highlighting the existing knowledge gaps in this field. CPP, conditioned place preference; NC, no change; N/A, not assessed. PSYCHEDELIC MEDICINE AND ZEBRAFISH 383 adult zebrafish.82–85 Some phenotypes of MK-801 exposure, New avenues of hallucinogenic medicine including hyperlocomotion and decreased performance in memory tasks, can be reversed by drugs (e.g., As classic serotonergic hallucinogens and dissociatives and ).86–88 Recently, antianxiety and have shown promise in treating psychiatric illnesses, more -like effects of MK-801 have also been reported clinical and preclinical research is needed to determine optimal in adult zebrafish.89,90 drugs, exposure paradigms, and doses for maximum effec- The j-opioid agonist salvinorin A, the principal psycho- tiveness. The zebrafish is exceptionally well suited to high- active component of S. divinorum, displays complex effects throughput drug screening and drug discovery.122–125 in both zebrafish and humans. Salvinorin A treatment sig- Additionally, neuropsychiatric disorders that may benefit from nificantly increased swimming activity and conditioned place hallucinogenic therapy (e.g., anxiety, depression, and addic- preference in adult zebrafish, an effect that was eliminated by tion) have been successfully modeled in zebrafish, evoking either j-opioid or CB1 antagonism.91 Salvinorin A also distinct behavioral profiles.126–134 Therefore, future studies modulates anxiety-like behavior in the NTT in a dose- should investigate the ability of hallucinogenic drugs to re- dependent manner92 but does not alter whole-body cortisol or verse or attenuate symptoms of anxiety and in brain c-fos expression (unpublished data). zebrafish. Indeed, the classic serotonergic hallucinogens, LSD and mescaline, elicit anxiolytic effects in wild-type treatment- Deliriant hallucinogens naive adult zebrafish.51,53 Testing of multiple drugs may elu- cidate differences in the efficacy of therapeutic effects between Deliriant-type hallucinogenic drugs are notably under- certain hallucinogens that act on similar receptor systems, studied in zebrafish models compared to both classic seroto- thereby offering insight into which drugs might be more useful nergic hallucinogens and dissociatives. Atropine reliably in the clinical setting. For example, psilocybin has been more affects cholinergic neuronal activity in the zebrafish telen- often utilized in clinical trials than other 5HT in cephalon,93 whereas scopolamine induces robust deficits in 2A recent years, possibly due to the stigma attached to LSD and learning and memory in adult zebrafish in numerous tests, mescaline use.2 However, LSD and mescaline are more ef- including Y-maze and aversive memory tasks.85,94–96 fective than psilocybin at eliciting anxiolysis in zebrafish models (although the difficult task of hallucinogenic dose ti- Utility of Zebrafish Models in Psychedelic tration may confound these studies),51,53,54 and the positive Medicine and Toxicology Research affective states induced by hallucinogenic drugs may be cru- cial for understanding their clinically relevant effects.39,135–137 Zebrafish models of complex Along with well-known psychedelic drugs, a number of re- neurobehavioral phenomena cently synthesized hallucinogenic agents have been discovered, The emerging field of psychedelic medicine has grown but not fully characterized in vivo.138,139 Here again, the nature exponentially in the past decade. However, the years of re- of zebrafish is beneficial for rapidly determining the behavioral strictive drug laws and insufficient clinical and preclinical and biochemical effects of numerous psychoactive compounds. research funding have exacted a toll on the current state of Some of these drugs may exhibit properties not possessed by hallucinogenic research.2,3,6 Recent calls to inform drug currently available psychedelics and therefore may be both policy utilizing a rational scale of harm have not resulted in novel and clinically relevant. Additionally, pharmacological policy change at the present juncture, despite the mounting and genomic methods in zebrafish can determine the precise evidence of potentially useful effects of hallucinogens.1 sites of action of novel drugs.112,125,134 In line with this, zeb- Although zebrafish are traditionally underutilized compared rafish models can help solve some remaining questions con- to rodents in neuroscience research, this model organism cerning hallucinogenic pharmacology. For example, LSD has a possesses attributes crucial for the examination of complex time-dependent dopaminergic component that affects locomo- neuropsychiatric phenomena.18 For example, the structure of tor activity and, possibly, anxiety-like behavior in rodents.140– zebrafish systems has been well studied, with 143 The ease of multiple dosing regimens in zebrafish, as well as serotonergic, opioidergic, glutamatergic, histaminergic, cho- the ability to measure metabolites in conjunc- linergic, and monoaminergic systems showing a high degree tion with behavior,75,144,145 may allow for a more thorough of complexity and homology to humans and other mam- mechanistic dissection of the temporal effects of hallucinogens mals.97–109 In addition, the zebrafish genome has been well than is currently possible with rodent and human models. characterized,110–113 and genome- and epigenome-wide ap- Additionally, the effects of hallucinogenic drugs on neu- proaches have been successfully utilized in this model.114,115 roendocrine and brain gene expression profiles may offer Zebrafish behavior has been comprehensively evaluated re- valuable insights into the underlying biology of psychedelic cently,56,116,117 and novel methods (such as three-dimensional states. Notably, for example, ketamine decreases mRNA video tracking) continue to advance our understanding of their levels of the phox2b transcription factor (when ketamine is complex phenotypes.52,118,119 Despite the invaluable contri- administered alone) and the histone deacetylase sirt1 (when butions of rodent models to hallucinogenic research,120 more ketamine is administered in hypoxic conditions) in adult preclinical work utilizing a wider breadth of model organisms zebrafish.67 The ability of hallucinogens to affect epigenetic is needed to determine the full extent of both the therapeutic factors and transcriptional enzymes should therefore be fur- benefits and toxicological risks associated with psychedelic ther examined in zebrafish and other aquatic models, as this therapy. The availability, high-throughput nature, and cost- may contribute to the well-known ability of psychedelics to benefit of zebrafish make this model attractive for screening evoke long-lasting changes in behavior (Fig. 2). compounds for behavioral effects, pharmacological efficacy, Another interesting topic relevant to hallucinogenic drug and toxicological properties.18,54,116,117,121 research is the concept of ‘‘’’, referring to the 384 KYZAR AND KALUEFF

FIG. 2. Zebrafish models to ad- vance psychedelic medicine and hallucinogenic drug discovery.

drug user’s own mindset and the environmental conditions zebrafish models can be valuable, and adult zebrafish have during drug exposure, respectively.146,147 As already men- been used to test potential toxic effects of high-dose halluci- tioned, zebrafish possess the necessary affective circuitry and nogen exposure in preliminary studies.152 The high-throughput robust behavioral responses to study the underlying neuro- nature of zebrafish screens means that large numbers of doses biology of ‘‘set and setting.’’ and many different drug–drug interactions can be rapidly tes- ted for deleterious effects in aquatic models.123 One potential concern is (SS), a life-threatening condi- Toxicology and safety testing of hallucinogens tion clinically defined by agitation, tachycardia, , using zebrafish models muscular rigidity, twitching, and diaphoresis. SS occurs when Although hallucinogens show promise in early clinical tri- serotonergic drugs (usually multiple drugs taken simulta- als for psychiatric disorders, valid concerns remain about the neously or in rapid succession) or other agents cause an ab- use of potent mind-altering compounds in therapy and re- errant increase in extracellular serotonin levels.153,154 SS has search.4 Hallucinogenic drugs do not usually elicit addictive- been replicated in zebrafish models,121 and future studies can like behaviors (i.e., compulsive drug seeking or withdrawal), utilize these models to determine the SS-evoking potential of such as alcohol, nicotine, , and .1 However, a hallucinogens, particularly when combined with other SS- vivid reexperiencing of the hallucinogen ‘‘high’’ occurring inducing agents. As poly-drug use (including long after the drug has left the user’s system, known as use) is prevalent in many populations,155–158 care should be hallucinogen-persisting perceptual disorder (Fig. 3), does taken to identify potential contraindications in preclinical rarely occur and can be serious in nature.148,149 Additionally, zebrafish screens to avoid issues in clinical practice. Given the powerful new synthetic hallucinogens, such as the N-o- teratogenic and neurotoxic effects of ketamine in zebrafish methoxybenzyl derivatives (NBOMe class of drugs), can be larvae,62,63 further research should also investigate the poten- fatal at high doses, and suicidality has been reported after tial neurodevelopmental effects of fetal drug exposure if psy- NBOMe ingestion.150,151 However, many reported adverse chedelic medicine is to reach wider audiences. effects of recreational hallucinogen use have not been ob- Finally, zebrafish may contribute to evaluating novel un- served in research settings, possibly due to the more controlled foreseen side effects of hallucinogenic therapy. For example, atmosphere and lack of precipitating factors, such as poor dos- ibogaine has gained an underground following for its anti- ing or use of other drugs.4,32 addictive effects, but major concerns have been raised follow- The risk of adverse effects and toxicological concerns ing fatalities seemingly resulting from ibogaine therapy.159–163 associated with hallucinogens is another domain in which Recent research suggests that ibogaine inhibits specific cardiac PSYCHEDELIC MEDICINE AND ZEBRAFISH 385

FIG. 3. Summary of hallucino- genic drug effects in humans and relevant phenotypes zebrafish.

channels, contributing to its ability to cause sudden cardiac proper discarding of this chemical waste, investigators face arrest and, in some cases, death.164–166 Ibogaine toxicity high- legal challenges and restrictions in handling even small lights the need for preclinical studies to examine the effects of amounts of diluted volumes of a Schedule I drug that are hallucinogenic drugs on organs other than the brain. Interest- greater than the legal ramifications of handling, for example, ingly, noribogaine (12-hydroxyibogamine), the stable and less- cocaine, a Schedule II drug.6 The solution to minimize such toxic metabolite of ibogaine,167,168 has shown therapeutic extreme situations may include declassifying hallucinogenic potential in both preclinical and clinical studies of affective substances or placing them under new research-specific disorders and .169–171 Noribogaine has a com- scheduling if used for biomedical studies in approved re- plex pharmacology, which includes the inhibition of the a3/a7 search facilities.1,2,6 nicotinic receptor subunits, noncompetitive inhi- bition of serotonin reuptake, and j- agonism.171 Conclusion Pilot studies with noribogaine in zebrafish have revealed re- duced novelty stress and anxiety-like behavior in the NTT and As psychedelic medicine moves into a new era of growth, an attenuation of -related phenotypes.171,59 preclinical models are becoming critical to better understand Partially overlapping with ibogaine action in zebrafish, these the benefits and potential detriments of hallucinogenic drug findings support the therapeutic potential of noribogaine for therapy.6 In this study, we have outlined the recent progress treating affective disorders comorbid with substance abuse. of treatment for various psychiatric disor- Noribogaine may be less capable of producing the adverse ef- ders, as well as the effects of hallucinogenic drugs on zeb- fects associated with ibogaine and may therefore represent a rafish physiology and behavior. Zebrafish and other aquatic safer alternative for development.168–170 The sen- models can prove particularly useful in testing hallucinogenic sitivity of zebrafish to the acute behavioral effects of nor- drugs for therapeutic efficacy, utilizing high-throughput ibogaine171 further highlights their utility for innovative screening for identifying novel hallucinogenic compounds, hallucinogenic drug research and development. Using zebrafish and rapidly determining adverse and/or toxicological effects and other model organisms to study the toxicology and phar- (Fig. 3). Collectively, this can foster more active utilization of macology of psychedelic drugs (Fig. 2) will better inform cli- zebrafish models to develop successful treatments for brain nicians and researchers of potential side effects in human disorders. populations, maximizing while minimizing risk. Finally, it is important to consider existing regulations and Acknowledgments drug laws that may impede basic and translational research into hallucinogens, particularly when using aquatic in vivo A.V.K. is the chair of ZNRC. His research is supported by models.6,172 For example, as many hallucinogens are Sche- the Guangdong Ocean University, the St. Petersburg State dule I drugs, any volume of drug-containing water (even in University internal grant 1.38.201.2014, and the Ural Federal very small doses) becomes a controlled Schedule I sub- University (the government of Russian Federation Act 211, stance.6 In addition to unclear or missing guidelines for contract 02-A03.21.0006). 386 KYZAR AND KALUEFF

Disclosure Statement 22. Heffter A. Ueber pellote. Naunyn Schmiedebergs Arch Pharmacol 1898;40:385–429. No competing financial interests exist. 23. Hofmann A: Discoveries in Biological Psychiatry. 1970, pp. 91–106. References J. B. Lippincott Co., Philadelphia, PA. 1. Nutt D, King LA, Saulsbury W, Blakemore C. Develop- 24. Hofmann A, Frey A, Ott H, Petrzilka T, Troxler F. Kon- ment of a rational scale to assess the harm of drugs of stitutionsaufkla¨rung und synthese von psilocybin. Ex- potential misuse. Lancet 2007;369:1047–1053. perientia 1958;14:397–399. 2. Nutt DJ, King LA, Nichols DE. Effects of Schedule I drug 25. Sherwood JN, Stolaroff MJ, Harman WW. The psyche- laws on neuroscience research and treatment innovation. delic experience—a new concept in psychotherapy. J Nat Rev Neurosci 2013;14:577–585. Neuropsychiatr 1962;4:69–80. 3. Nutt D. Illegal drugs laws: clearing a 50-year-old obstacle 26. Pahnke WN, Kurland AA, Unger S, Savage C, Grof S. to research. PLoS Biol 2015;13:e1002047. The experimental use of psychedelic (LSD) psychother- 4. Johnson M, Richards W, Griffiths R. Human hallucinogen apy. Int Z Klin Pharmakol Ther Toxikol 1971;4:446–454. research: guidelines for safety. J Psychopharmacol 2008; 27. Oram M. Efficacy and Enlightenment: LSD Psychother- 22:603–620. apy and the Drug Amendments of 1962. J Hist Med Allied 5. Sessa B. Shaping the renaissance of psychedelic research. Sci 2014;69:221–250. Lancet 2012;380:200–201. 28. Kurland AA, Unger S, Shaffer JW, Savage C. Psychedelic 6. Stewart AM, Kalueff AV. Controlled substances and in- therapy utilizing LSD in the treatment of the alcoholic novation of biomedicine: a preclinical perspective. Nat patient: a preliminary report. Am J Psychiatry 1967;123: Rev Neurosci 2013;14:877. 1202–1209. 7. Nichols DE. Hallucinogens. Pharmacol Ther 2004;101: 29. Grof S, Goodman L, Richards W, Kurland A. LSD- 131–181. assisted psychotherapy in patients with terminal cancer. 8. Halberstadt AL. Recent advances in the neuropsycho- Int Pharmacopsychiat 1973; 8:129–144. pharmacology of serotonergic hallucinogens. Behav Brain 30. Harman WW, McKim RH, Mogar RE, Fadiman J, Sto- Res 2015;277:99–120. laroff MJ. Psychedelic agents in creative problem-solving: 9. Glennon RA. Arylalkylamine drugs of abuse: an overview a pilot study. Psychol Rep 1966;19:211–227. of drug discrimination studies. Pharmacol Biochem Behav 31. Rhead JC. Psychedelic medicine: new evidence for hallu- 1999;64:251–256. cinogenic substances as treatments. J Psychoactive Drugs 10. Grob CS, de Rios MD, Rosner R. Hallucinogens and re- 2014;46:78–83. lated compounds. Clin Handbook Adolesc Addict 2013; 32. Bogenschutz MP, Johnson MW. Classic hallucinogens in 213–222. the treatment of . Prog Neuro Psychopharmacol 11. Vollenweider FX, Geyer MA. A systems model of altered Biol Psychiatry 2016;64:250–258. : integrating natural and drug-induced psy- 33. Berman RM, et al. Antidepressant effects of ketamine in choses. Brain Res Bull 2001;56:495–507. depressed patients. Biol Psychiatry 2000;47:351–354. 12. Roth BL, et al. Salvinorin A: a potent naturally occurring 34. Machado-Vieira R, Salvadore G, DiazGranados N, Zarate nonnitrogenous kappa opioid selective agonist. Proc Natl CA. Ketamine and the next generation of Acad Sci U S A 2002;99:11934–11939. with a rapid onset of action. Pharmacol Ther 2009;123: 13. Mahler DA. Anticholinergic poisoning from Jimson weed. 143–150. JACEP 1976;5:440–442. 35. Duman RS, Li N, Liu RJ, Duric V, Aghajanian G. Sig- 14. Ceha LJ, Presperin C, Young E, Allswede M, Erickson T. naling pathways underlying the rapid antidepressant ac- Anticholinergic toxicity from nightshade berry poisoning tions of ketamine. Neuropharmacology 2012;62:35–41. responsive to physostigmine. J Emerg Med 1997;15:65–69. 36. Li N, et al. mTOR-dependent synapse formation underlies 15. Popik P, Skolnick P. Pharmacology of Ibogaine and the rapid antidepressant effects of NMDA antagonists. Ibogaine-Related Alkaloids. Alkaloids 1999;52:197–231. Science 2010;329:959–964. 16. Mah SJ, Tang Y, Liauw PE, Nagel JE, Schneider AS. 37. Newport DJ, et al. Ketamine and other NMDA antago- Ibogaine acts at the nicotinic to in- nists: early clinical trials and possible mechanisms in hibit catecholamine release. Brain Res 1998;797:173–180. depression. Am J Psychiatry 2015;172:950–966. 17. Maciulaitis R, Kontrimaviciute V, Bressolle FMM, Brie- 38. Krebs TS, Johansen PØ. Lysergic acid diethylamide dis V. Ibogaine, an anti-addictive drug: pharmacology and (LSD) for alcoholism: meta-analysis of randomized con- time to go further in development. A narrative review. trolled trials. J Psychopharmacol 2012;26:994–1002. Hum Exp Toxicol 2008;27:181–194. 39. Bogenschutz MP, et al. Psilocybin-assisted treatment for 18. Neelkantan N, et al. Perspectives on zebrafish models of : a proof-of-concept study. J Psycho- hallucinogenic drugs and related psychotropic com- pharmacol 2015;29:289–299. pounds. ACS Chem Neurosci 2013;4:1137–1150. 40. Johnson MW, Garcia-Romeu A, Cosimano MP, Griffiths 19. El-Seedi HR, De Smet PAGM, Beck O, Possnert G, Bruhn RR. Pilot study of the 5-HT2AR agonist psilocybin in the JG. Prehistoric peyote use: analysis and radio- treatment of addiction. J Psychopharmacol 2014;28: dating of archaeological specimens of Lophophora 983–992. from Texas. J Ethnopharmacol 2005;101:238–242. 41. Tupper KW, Wood E, Yensen R, Johnson MW. Psyche- 20. McKenna DJ. Ayahuasca and human destiny. J Psy- delic medicine a re-emerging therapeutic paradigm. choactive Drugs 2005;37:231–234. CMAJ 2015;187:1054–1059. 21. Winkelman M. Drug tourism or spiritual ? Aya- 42. Gasser P, et al. Safety and efficacy of lysergic acid huasca seekers in Amazonia. J Psychoactive Drugs 2005; diethylamide-assisted psychotherapy for anxiety associated 37:209–218. with life-threatening diseases. J Nerv Ment Dis 2014;00:1–8. PSYCHEDELIC MEDICINE AND ZEBRAFISH 387

43. Gasser P, Kirchner K, Passie T. LSD-assisted psycho- MAPK (ERK) activity in zebrafish embryos. Reprod therapy for anxiety associated with a life-threatening Toxicol 2012;33:205–212. disease: a qualitative study of acute and sustained sub- 61. Cuevas E, et al. Acetyl l-carnitine protects motor jective effects. J Psychopharmacol 2015;29:57–68. and Rohon-Beard sensory neurons against ketamine- 44. Grob CS, et al. Pilot study of psilocybin treatment for induced in zebrafish embryos. Neurotoxicol anxiety in patients with advanced-stage cancer. Arch Gen Teratol 2013;39:69–76. Psychiatry 2011;68:71–78. 62. Kanungo J, Cuevas E, Ali SF, Paule MG. Ketamine in- 45. Mithoefer MC, Wagner MT, Mithoefer AT, Jerome L, duces motor toxicity and alters neurogenic and Doblin R. The safety and efficacy of –3,4-methylenedio- proneural gene expression in zebrafish. J Appl Toxicol xymethamphetamine-assisted psychotherapy in subjects 2013;33:410–417. with chronic, treatment-resistant posttraumatic stress dis- 63. Fe´lix LM, Antunes LM, Coimbra AM. Ketamine NMDA order: the first randomized controlled pilot study. J Psy- receptor-independent toxicity during zebrafish (Danio re- chopharmacol 2011;25:439–452. rio) embryonic development. Neurotoxicol Teratol 2014; 46. Mithoefer MC, et al. Durability of improvement in post- 41:27–34. traumatic stress disorder symptoms and absence of 64. Robinson BL, Dumas M, Paule MG, Ali SF, Kanungo J. harmful effects or drug dependency after 3,4-methyl- Opposing effects of ketamine and acetyl l -carnitine on the enedioxymethamphetamine-assisted psychotherapy: a serotonergic system of zebrafish. Neurosci Lett prospective long-term follow-up study. J Psychopharma- 2015;607:17–22. col 2013;27:28–39. 65. Trickler WJ, et al. Ketamine attenuates cytochrome p450 47. Oehen P, Traber R, Widmer V, Schnyder U. A randomized, aromatase gene expression and estradiol-17b levels in zeb- controlled pilot study of MDMA (– 3,4-Methylenedio- rafish early life stages. J Appl Toxicol 2014;34:480–488. xymethamphetamine)-assisted psychotherapy for treatment 66. Riehl R, et al. Behavioral and physiological effects of of resistant, chronic Post-Traumatic Stress Disorder (PTSD). acute ketamine exposure in adult zebrafish. Neurotoxicol J Psychopharmacol 2013;27:40–52. Teratol 2011;33:658–667. 48. Nichols DE. Differences between the 67. Zakhary SM, et al. A behavioral and molecular analysis of of MDMA, MBDB, and the classic hallucinogens. Iden- ketamine in zebrafish. Synapse 2011;65:160–167. tification of a new therapeutic class: entactogens. J Psy- 68. De Campos EG, Bruni AT, De Martinis BS. Ketamine choact Drugs 1986;18:305–313. induces anxiolytic effects in adult zebrafish: a multivariate 49. Oberlender R, Nichols DE. Drug discrimination studies statistics approach. Behav Brain Res 2015;292:537–546. with MDMA and . 69. Hetzler BE, Wautlet BS. Ketamine-induced locomotion in (Berl) 1988;95:71–76. rats in an open-field. Pharmacol Biochem Behav 50. Vollenweider FX. Brain mechanisms of hallucinogens and 1985;22:653–655. entactogens. Dialogues Clin Neurosci 2001;3:265–279. 70. Murata S, Kawasaki K. Common and uncommon beha- 51. Grossman L, et al. Characterization of behavioral and vioural effects of antagonists for different modulatory endocrine effects of LSD on zebrafish. Behav Brain Res sites in the NMDA receptor/channel complex. Eur J 2010;214:277–284. Pharmacol 1993;239:9–15. 52. Green J, et al. Automated high-throughput neuropheno- 71. Fan N, et al. Development of a checklist of short-term and typing of zebrafish social behavior. J Neurosci Methods long-term psychological symptoms associated with keta- 2012;210:266–271. mine use. Shanghai Arch Psychiatry 2015;27:186–194. 53. Kyzar EJ, et al. Effects of hallucinogenic agents mesca- 72. Ben Fredj N, et al. Synaptic activity and activity- line and phencyclidine on zebrafish behavior and physi- dependent competition regulates axon arbor maturation, ology. Prog Neuropsychopharmacol Biol Psychiatry growth arrest, and territory in the retinotectal projection. J 2012;37:194–202. Neurosci 2010;30:10939–10951. 54. Kyzar EJ:. Novel Models to Study Hallucinogenic Drug 73. Schmidt JT, Buzzard M, Borress R, Dhillon S. MK801 Action: Behavioral and Physiological Effects of Psilocy- increases retinotectal arbor size in developing zebrafish bin and Phencyclidine in Adult Zebrafish. Tulane Uni- without affecting kinetics of branch elimination and ad- versity, New Orleans, LA, 2012. dition. J Neurobiol 2000;42:303–314. 55. Stewart A, et al. Behavioral effects of MDMA (‘ecstasy’) 74. Schmidt JT, Fleming MR, Leu B. Presynaptic protein on adult zebrafish. Behav Pharmacol 2011;22:275–280. kinase C controls maturation and branch dynamics of 56. Kalueff AV, et al. Towards a comprehensive catalog of developing retinotectal arbors: possible role in activity- zebrafish behavior 1.0 and beyond. Zebrafish 2013;10: driven sharpening. J Neurobiol 2004;58:328–340. 70–86. 75. Liu X, et al. Strain-dependent differential behavioral re- 57. Cachat J, et al. Unique and potent effects of acute ibo- sponses of zebrafish larvae to acute MK-801 treatment. gaine on zebrafish: the developing utility of novel aquatic Pharmacol Biochem Behav 2014;127:82–89. models for hallucinogenic drug research. Behav Brain Res 76. Chen J, et al. The Behavioral and pharmacological actions 2013;236:258–269. of NMDA receptor antagonism are conserved in zebrafish 58. Glick SD, Maisonneuve IM. Development of novel larvae. Int J Comp Psychol 2010;23:82–90. for drug addiction. The legacy of an African 77. Andersson MA˚ , Ek F, Olsson R. Using visual lateraliza- shrub. Ann N Y Acad Sci 2000;909:88–103. tion to model learning and memory in zebrafish larvae. Sci 59. Li X, Kalueff AV, Maillet EL. Anxiolytic effects of nor- Rep 2015;5:8667. ibogaine on novelty stress in rats and zebrafish. Soc Toxicol 78. Swain HA, Sigstad C, Scalzo FM. Effects of Abstract #1661, P548, 2016. (MK-801) on circling behavior, swimming activity, and 60. Kanungo J, Cuevas E, Ali SF, Paule MG. L-Carnitine place preference in zebrafish (Danio rerio). Neurotoxicol rescues ketamine-induced attenuated heart rate and Teratol 2004;26:725–729. 388 KYZAR AND KALUEFF

79. Tran S, Muraleetharan A, Fulcher N, Chatterjee D, Gerlai 97. Alsop D, Vijayan M. The zebrafish stress axis: molecular R. MK-801 increases locomotor activity in a context- fallout from the teleost-specific genome duplication event. dependent manner in zebrafish. Behav Brain Res 2016; Gen Comp Endocrinol 2009;161:62–66. 296:26–29. 98. Rodriguez-Martin I, de Velasco EMF, Rodriguez RE. 80. Sison M, Gerlai R. Behavioral performance altering ef- Characterization of -binding sites in zebrafish fects of MK-801 in zebrafish (Danio rerio). Behav Brain brain. Neurosci Lett 2007;413:249–254. Res 2011;220:331–337. 99. De Velasco EMF, Law PY, Rodrı´guez RE. Mu opioid 81. Menezes FP, Kist LW, Bogo MR, Bonan CD, Da Silva RS. receptor from the zebrafish exhibits functional character- Evaluation of age-dependent response to NMDA receptor istics as those of mammalian mu opioid receptor. Zebra- antagonism in zebrafish. Zebrafish 2015;12:137–143. fish 2009;6:259–268. 82. Xu X, Scott-Scheiern T, Kempker L, Simons K. Active 100. Pinal-Seoane N, et al. Characterization of a new duplicate avoidance conditioning in zebrafish (Danio rerio). Neu- delta-opioid receptor from zebrafish. J Mol Endocrinol robiol Learn Mem 2007;87:72–77. 2006;37:391–403. 83. Blank M, Guerim LD, Cordeiro RF, Vianna MR. M. A 101. Lillesaar C. The serotonergic system in fish. J Chem one-trial inhibitory avoidance task to zebrafish: rapid ac- Neuroanat 2011;41:294–308. quisition of an NMDA-dependent long-term memory. 102. Maximino C, et al. The serotonergic system of zebrafish: Neurobiol Learn Mem 2009;92:529–534. genomics, neuroanatomy and neuropharmacology. Ser- 84. Sison M, Gerlai R. Associative learning performance is otonin Biosynthesis Regul Heal Implic 2013;53–67. impaired in zebrafish (Danio rerio) by the NMDA-R an- 103. Panula P, et al. The comparative neuroanatomy and neu- tagonist MK-801. Neurobiol Learn Mem 2011;96:230– rochemistry of zebrafish CNS systems of relevance to 237. human neuropsychiatric diseases. Neurobiol Dis 85. Cognato GDP, et al. Y-Maze memory task in zebrafish 2010;40:46–57. (Danio rerio): the role of glutamatergic and cholinergic 104. Panula P, et al. Modulatory neurotransmitter systems and systems on the acquisition and consolidation periods. behavior: towards zebrafish models of neurodegenerative Neurobiol Learn Mem 2012;98:321–328. diseases. Zebrafish 2006;3:235–247. 86. Seibt KJ, et al. Antipsychotic drugs prevent the motor 105. Kaslin J, Panula P. Comparative anatomy of the hista- hyperactivity induced by MK-801 in minergic and other aminergic systems in zebrafish (Danio zebrafish (Danio rerio). Behav Brain Res 2010;214:417– rerio). J Comp Neurol 2001;440:342–377. 422. 106. Sundvik M, Panula P. Organization of the histaminergic 87. Seibt KJ, et al. Antipsychotic drugs reverse MK-801- system in adult zebrafish (Danio rerio) brain: neuron induced cognitive and social interaction deficits in zeb- number, location, and cotransmitters. J Comp Neurol rafish (Danio rerio). Behav Brain Res 2011;224:135–139. 2012;520:3827–3845. 88. Seibt KJ, et al. MK-801 alters Na+,K+-ATPase activity 107. Kastenhuber E, Kratochwil CF, Ryu S, Schweitzer J, and oxidative status in zebrafish brain: reversal by anti- Driever W. Genetic dissection of dopaminergic and nor- psychotic drugs. J Neural Transm 2012;119:661–667. adrenergic contributions to catecholaminergic tracts in 89. Herculano AM, Puty B, Miranda V, Lima MG, Maximino early larval zebrafish. J Comp Neurol 2010;518:439–458. C. Interactions between serotonin and glutamate–nitric 108. Tay TL, Ronneberger O, Ryu S, Nitschke R, Driever W. oxide pathways in zebrafish scototaxis. Pharmacol Bio- Comprehensive catecholaminergic projectome analysis re- chem Behav 2015;129:97–104. veals single-neuron integration of zebrafish ascending and 90. Bohrer R, Maria A, Denise C. The role of purinergic and descending dopaminergic systems. Nat Commun 2011;2:171. dopaminergic systems on MK-801-induced antidepressant 109. Rink E, Wullimann MF. Development of the catechol- effects in zebrafish. Pharmacol Biochem Behav 2015; aminergic system in the early zebrafish brain: an immu- 139:149–157. nohistochemical study. Dev Brain Res 2002;137:89–100. 91. Braida D, et al. Hallucinatory and rewarding effect of 110. Postlethwait J, Amores A: The zebrafish genome. Meth- salvinorin A in zebrafish: Kappa-opioid and CB1- ods in Cell Biology 1998, pp. 149–163. cannabinoid receptor involvement. Psychopharmacology 111. Woods IG, et al. A comparative map of the zebrafish (Berl) 2007;190:441–448. genome. Genome Res 2000;10:1903–1914. 92. Collins C: The Zebrafish (Danio rerio) as a Highly Sen- 112. Schier AF. Genomics: zebrafish earns its stripes. Nature sitive Tool for Screening Hallucinogenic Drug Action: 2013;496:443–444. Lessons from Mescaline and Salvinorin A. Tulane Uni- 113. Kettleborough RNW, et al. A systematic genome-wide versity, New Orleans, LA, 2012. analysis of zebrafish protein-coding gene function. Nature 93. Park E, Lee Y, Kim Y, Lee CJ. Cholinergic modulation of 2013;496:494–497. neural activity in the telencephalon of the zebrafish. 114. Aday AW, Zhu LJ, Lakshmanan A, Wang J, Lawson ND. Neurosci Lett 2008;439:79–83. Identification of cis regulatory features in the embryonic 94. Kim YH, Lee Y, Kim D, Jung MW, Lee CJ. Scopolamine- zebrafish genome through large-scale profiling of induced learning impairment reversed by physostigmine H3K4me1 and H3K4me3 binding sites. Dev Biol in zebrafish. Neurosci Res 2010;67:156–161. 2011;357:450–462. 95. Richetti SK, et al. Quercetin and rutin prevent 115. Harrison MRM, Georgiou AS, Spaink HP, Cunliffe VT. scopolamine-induced memory impairment in zebrafish. The epigenetic regulator Histone Deacetylase 1 promotes Behav Brain Res 2011;217:10–15. transcription of a core neurogenic programme in zebrafish 96. Bortolotto JW, de Melo GM, Cognato GDP, Vianna embryos. BMC Genomics 2011;12:24. MRM, Bonan CD. Modulation of adenosine signaling 116. Kalueff AV, Stewart AM, Gerlai R. Zebrafish as an prevents scopolamine-induced cognitive impairment in emerging model for studying complex brain disorders. zebrafish. Neurobiol Learn Mem 2015;118:113–119. Trends Pharmacol Sci 2014;35:63–75. PSYCHEDELIC MEDICINE AND ZEBRAFISH 389

117. Spence R, Gerlach G, Lawrence C, Smith C. The behav- 138. Shulgin AT, Shulgin A: PIHKAL: A Chemical Love iour and ecology of the zebrafish, Danio rerio. Biol Rev Story. Transform Press, Berkeley, CA, 1991. 2008;83;13–34. 139. Shulgin AT, Shulgin A: TIHKAL: The Continuation. 118. Stewart AM, et al. A novel 3D method of locomotor Transform Press, Berkeley, CA, 1997. analysis in adult zebrafish: implications for automated 140. Marona-Lewicka D, Thisted RA, Nichols DE. Distinct detection of CNS drug-evoked phenotypes. J Neurosci temporal phases in the behavioral pharmacology of LSD: Methods 2015;255:66–74. D2 receptor-mediated effects in the rat and 119. Cachat J, et al. Three-dimensional neurophenotyping of implications for . Psychopharmacology (Berl) adult zebrafish behavior. PLoS One 2011;6. 2005;180:427–435. 120. Halberstadt AL, Geyer MA. Characterization of the head- 141. Marona-Lewicka D, Nichols DE. Further evidence that the twitch response induced by hallucinogens in mice: detection delayed temporal dopaminergic effects of LSD are medi- of the behavior based on the dynamics of head movement. ated by a mechanism different than the first temporal phase Psychopharmacology (Berl) 2013;227:727–739. of action. Pharmacol Biochem Behav 2007;87:453–461. 121. Stewart AM, et al. Perspectives on experimental models 142. Marona-Lewicka D, Chemel B, Nichols D. Dopamine D4 of serotonin syndrome in zebrafish. Neurochem Int receptor involvement in the discriminative stimulus effects 2013;62:893–902. in rats of LSD, but not the hallucinogen 122. Parng C, Seng WL, Semino C, McGrath P. Zebrafish: a DOI. Psychopharmacology (Berl) 2009;203:265–277. preclinical model for drug screening. Assay Drug Dev 143. Kyzar EJ, Stewart AM, Kalueff AV. Effects of LSD on Technol 2002;1:41–48. grooming behavior in heterozygous 123. Rubinstein AL. Zebrafish assays for drug toxicity (Sert) mice. Behav Brain Res 2015;296:47–52. screening. Expert Opin Drug Metab Toxicol 2006;2:231– 144. Eddins D, Petro A, Williams P, Cerutti DT, Levin ED. 240. Nicotine effects on learning in zebrafish: the role of do- 124. Murphey RD, Zon LI. Small molecule screening in the paminergic systems. Psychopharmacology (Berl) 2009; zebrafish. Methods 2006;39:255–261. 202:103–109. 125. Zon LI, Peterson RT. In vivo drug discovery in the zeb- 145. Chatterjee D, Gerlai R. High precision liquid chroma- rafish. Nat Rev Drug Discov 2005;4:35–44. tography analysis of dopaminergic and serotoninergic re- 126. Stewart A, et al. Modeling anxiety using adult zebrafish: sponses to acute alcohol exposure in zebrafish. Behav a conceptual review. Neuropharmacology 2012;62: Brain Res 2009;200:208–213. 135–143. 146. Tupin JP. Drug, Set, and Setting: the Basis for Controlled 127. Kyzar E, et al. Behavioral effects of bidirectional modu- Intoxicant Use. Am J Psychiatry 1986;143:548–549. lators of brain monoamines and d-amphetamine 147. Eisner B. Set, setting, and matrix. J Psychoactive Drugs in zebrafish. Brain Res 2013;1527:108–116. 2011;29:213–216. 128. Stewart A, et al. Pharmacological modulation of anxiety- 148. Halpern J. Hallucinogen persisting disorder: like phenotypes in adult zebrafish behavioral models. Prog what do we know after 50 years? Drug Alcohol Depend Neuro Psychopharmacol Biol Psychiatry 2011;35:1421– 2003;69:109–119. 1431. 149. Hermle L, Simon M, Ruchsow M, Geppert M. 129. Stewart A, et al. Zebrafish models to study drug abuse- Hallucinogen-persisting perception disorder. Ther Adv related phenotypes. Rev Neurosci 2011;22:95–105. Psychopharmacol 2012;2:199–205. 130. Gerlai R, Ahmad F, Prajapati S. Differences in acute 150. Umemura Y, et al. Fatal 25I-NBOMe intoxication: a new alcohol-induced behavioral responses among zebrafish recreational risk. Acad Forensic Pathol 2015;5:91–97. populations. Alcohol Clin Exp Res 2008;32:1763–1773. 151. Suzuki J, Poklis JL, Poklis A. ‘My friend said it was good 131. Miller N, Greene K, Dydinski A, Gerlai R. Effects of LSD’: a suicide attempt following analytically confirmed nicotine and alcohol on zebrafish (Danio rerio) shoaling. 25I-NBOMe ingestion. J Psychoact Drugs 2014;46:379–382. Behav Brain Res 2013;240:192–196. 152. Kalueff AV, et al. Zebrafish neurobehavioral phenomics 132. Pan Y, Kaiguo M, Razak Z, Westwood JT, Gerlai R. for aquatic neuropharmacology and toxicology research. Chronic alcohol exposure induced gene expression chan- Aquat Toxicol 2016;170:297–309. ges in the zebrafish brain. Behav Brain Res 2011;216:66– 153. Haberzettl R, Bert B, Fink H, Fox MA. Animal models of 76. the serotonin syndrome: a systematic review. Behav Brain 133. Tran S, Chatterjee D, Gerlai R. An integrative analysis of Res 2013;256:328–345. tolerance and withdrawal in zebrafish (Danio rerio). 154. Boyer EW, Shannon M. The serotonin syndrome. N Engl Behav Brain Res 2014;276:161–170. J Med 2005;352:1112–1120. 134. Klee EW, et al. Zebrafish: a model for the study of ad- 155. Kelly BC, Wells BE, Pawson M, LeClair A, Parsons JT. diction genetics. Hum Genet 2012;131:977–1008. Combinations of prescription drug misuse and illicit drugs 135. Kometer M, et al. Psilocybin biases facial recognition, among young adults. Addict Behav 2014;39:941–944. goal-directed behavior, and mood state toward positive 156. McCabe SE, Cranford JA, Morales M, Young A. Si- relative to negative through different seroto- multaneous and concurrent polydrug use of alcohol and nergic subreceptors. Biol Psychiatry 2012;72:898–906. prescription drugs: prevalence, correlates, and conse- 136. Carhart-Harris RL, et al. Neural correlates of the psy- quences. J Stud Alcohol 2006;67:529–537. chedelic state as determined by fMRI studies with psilo- 157. Jones JD, Mogali S, Comer SD. Polydrug abuse: a review cybin. Proc Natl Acad Sci U S A 2012;109:2138–2143. of opioid and combination use. Drug Al- 137. Carhart-Harris RL, et al. Implications for psychedelic- cohol Depend 2012;125:8–18. assisted psychotherapy: functional magnetic resonance 158. Kedia S, Sell MA, Relyea G. Mono- versus polydrug abuse imaging study with psilocybin. Br J Psychiatry 2012;200: patterns among publicly funded clients. Subst Abuse Treat 238–244. Prev Policy 2007;2:33. 390 KYZAR AND KALUEFF

159. Alper KR, Stajic´ M, Gill JR. Fatalities temporally associ- 170. Carnicella S, He DY, Yowell QV, Glick SD, Ron D. ated with the ingestion of ibogaine. J Forensic Sci 2012;57: Noribogaine, but not 18-MC, exhibits similar actions 398–412. as ibogaine on GDNF expression and ethanol self- 160. Maas U, Strubelt S. Fatalities after taking ibogaine in administration. Addict Biol 2010;15:424–433. addiction treatment could be related to sudden cardiac 171. Maillet EL. Noribogaine multitarget directed pharmacol- death caused by autonomic dysfunction. Med Hypotheses ogy: a canonical reciple for addiction and anxiety related 2006;67:960–964. disorders? In Abstracts of the 5th Regional ‘Stress and 161. Alper KR, Lotsof HS, Kaplan CD. The ibogaine medical Behavior’ ISBS Conference 2012, pp. 12–13. . J Ethnopharmacol 2008;115:9–24. 172. Nutt DJ, King LA, Nichols DE. New victims of current 162. Papadodima SA, Dona A, Evaggelakos CI, Goutas N, drug laws. Nat Rev Neurosci 2013;14:877. Athanaselis SA. Ibogaine related sudden death: a case 173. Mann KD, et al. Cardiac response to startle stimuli in report. J Forensic Leg Med 2013;20:809–811. larval zebrafish: sympathetic and parasympathetic com- 163. Vastag B. Addiction research. Ibogaine therapy: a ‘vast, ponents. Am J Physiol Regul Integr Comp Physiol uncontrolled experiment’. Science 2005;308:345–346. 2010;98:R1288–R1297. 164. Koenig X, Kovar M, Boehm S, Sandtner W, Hilber K. 174. Best JD, et al. Non-associative learning in larval zebrafish. Anti-addiction drug ibogaine inhibits hERG chan- 2008;33:1206–1215. nels: a cardiac arrhythmia risk. Addict Biol 2014;19: 237–239. Address correspondence to: 165. Koenig X, et al. Anti-addiction drug ibogaine inhibits Evan J. Kyzar, BS voltage-gated ionic currents: a study to assess the drug’s Department of Psychiatry, College of Medicine cardiac profile. Toxicol Appl Pharmacol University of Illinois at Chicago 2013;273:259–268. 166. Koenig X, Hilber K. The anti-addiction drug ibogaine 1601 West Taylor Street and the heart: a delicate relation. Molecules 2015;20: Chicago, IL 60612 2208–2228. E-mail: [email protected] 167. Glue P, et al. Ascending-dose study of noribogaine in healthy volunteers: pharmacokinetics, , Allan V. Kalueff, PhD safety, and tolerability. J Clin Pharmacol 2015;55:189–194. Research Institute for Marine Drugs and Nutrition 168. Kubiliene A, et al. Acute toxicity of ibogaine and nor- College of Food Science and Technology ibogaine. Medicina (Kaunas) 2008;44:984–988. Guangdong Ocean University 169. Baumann MH, Pablo JP, Ali SF, Rothman RB, Mash DC. Zhanjiang 524025 Noribogaine (12-Hydroxyibogamine): a biologically ac- China tive metabolite of the antiaddictive drug ibogaine. Ann NY Acad Sci 2000;914:354–368. E-mail: [email protected]