Schizophrenia Research 133 (2011) 264265

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Schizophrenia Research
journal homepage: www.elsevier.com/locate/schres

Letter to the Editor

Adrenochrome and related oxidative metabolites of catecholamines: Effects on dopamine neurons and receptor binding proles also screened for potential binding sites of adrenochrome through the National Institute of Mental Health's Psychoactive Drug Screening Program (Contract # HHSN-271-2008-00025-C). Injection of adrenochrome (48 mg/kg, i.v.) produced no signicant effect on DA cell ring rate (from 4.6 0.9 to 4.2 0.9 spikes/s, n = 6, p = 0.14, paired t-test), variability of ring (CV, from 51.8 9.7 to 49.3 5.6%, p = 0.71), and slow oscillation in ring (SO, relative spectral power between 0.4 and 1.2 Hz, from 0.48 0.19 to 0.36 0.11%, p = 0.81, Fig. 1AC). Adrenolutin (39 mg/kg, i.v., n = 6), a relatively stable derivative from adrenochrome, also had no signicant effect on ring rate (from 3.9 1.1 to 4.1 1.0 spikes/s, p = 0.67) and CV (from 42 4.0% to 46 8.2%, p = 0.55), and induced a small decrease in SO (from 0.23 0.04% to 0.18 0.04%, p = 0.04, Fig. 1C). In contrast, the DA precursor L-DOPA (50100 mg/kg, i.v., n = 9) consistently inhibited all three measures of DA cell activity (ring rate: from 5.6 0.3 to 4.3 0.4 spikes/s, p b 0.001; CV: from 59.6 8.1% to 41.3 7.0%, p b 0.001; SO: from 0.51 0.14% to 0.26 0.10%, p b 0.001, Fig. 1C). All effects were reversed by the D2 antagonist raclopride (50100 g/kg, i.v.). The psychotomimetic drug D-amphetamine (0.51.0 mg/kg, i.v.) also inhibited the ring rate (4.9 0.7 to 3.3 0.8 spikes/s, p = 0.001, n = 6), but it produced variable effects on CV and SO. Overall, CV and SO were unchanged (CV: from 43.8 6.3% to

Dear Editors, In 1954, Hoffer et al. reported that adrenochrome, an oxidative metabolite of epinephrine, was psychotomimetic in humans when administered intravenously (Hoffer et al., 1954). This and other observations led the authors to hypothesize that adrenochrome is overproduced and responsible for some symptoms in schizophrenia (see review by Mills, 2010). Hoffer et al. further proposed that vitamin B3 (niacin) might be therapeutic for schizophrenia since it may act as a methyl acceptor to decrease the conversion of norepinephrine to epinephrine, thus inhibiting adrenochrome synthesis. Although this transmethylation theory has received a great deal of criticism (Mills, 2010), the psychotomimetic effect of adrenochrome seems to have been conrmed (Schwarz et al., 1956; Grof et al., 1963). To test whether adrenochrome acts in part through central dopamine (DA) systems, we made single-cell recordings from DA neurons in the ventral tegmental area in chloral hydrate-anesthetized rats. We

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Fig. 1. Comparison of effects of adrenochrome, adrenolutin, L-DOPA, and D-amphetamine on DA neurons. A) Typical rate histogram (top, binwidth: 10 s) of a DA neuron and corresponding interspike interval (ISI) plots (bottom) showing that adrenochrome produced no signicant effect on the ring rate and induced only a transient decrease in the variability of ISIs at high doses. Each dot in the ISI plots represents one ISI. B) Right: segments of spike trains and corresponding smoothed rate histograms (binwidth: 50 ms) from the same neuron showing the presence of SO both under baseline conditions and after the last dose of adrenochrome. Left: autospectra of the rate histograms showing a lack of a signicant change in SO by adrenochrome. C) Bar graphs comparing effects of adrenochrome and adrenolutin with those of L-DOPA and D-amphetamine. Adrenochrome had no signicant effect on all three measures of DA cell activity, whereas adrenolutin produced a small suppression of SO. In contrast, L-DOPA inhibited all three measures, while D-amphetamine decreased the ring rate and slightly increased CV and SO. The changes in CV and SO induced by D-amphetamine, however, were statistically insignicant. *p = 0.04, ***p 0.001. 0920-9964/$ see front matter 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.schres.2011.08.015

Letter to the Editor

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45.4 6.1%, p = 0.847; SO: from 0.18 0.04% to 0.26 0.07%, p = 0.267, Fig. 1C). Similar results were previously obtained with cocaine (Zhou et al., 2006). L-DOPA and D-amphetamine inhibit DA cell ring through DAmediated feedback mechanisms. The lack of the inhibitory effect of adrenochrome and adrenolutin suggests that the two compounds are unlikely to have a signicant inuence on DA synthesis, release, or reuptake, nor do they directly bind to and stimulate DA receptors. Binding assays conrmed that the two compounds as well as two other catecholamine metabolites, 5,6-dihydroxyindole (DHI) and 5,6-dihydroxyindole-2-carboxylic acid (DHICA), lacked a signicant afnity for all ve DA receptor subtypes (D15) and the DA transporter. Amphetamine-like psychotomimetics also have an excitatory effect on DA neurons, which is in part mediated through adrenergic 1 receptors (Shi et al., 2000; Shi et al., 2004). Our recordings suggest that adrenochrome and adrenolutin do not mimic the effect. The two compounds as well as DHICA and DHI also showed no signicant afnity for the norepinephrine transporter and for most norepinephrine receptors screened, including 1 receptors. Adrenochrome, adrenolutin, and DHICA, however, displayed a low-micromole afnity for 2c receptors with Ki values of 2.1, 2.7 and 1.8 M, respectively. These values are on a par with the afnity of norepinephrine (1.3 M) and epinephrine (1.7 M) for the same receptor (Jasper et al., 1998). DA neurons have been shown to express 2c receptors (Rosin et al., 1996). Activation of 2c receptors reduces the hyperpolarizationactivated cation current, leading to an increased ring in DA cells (Inyushin et al., 2010). Whether adrenochrome and adrenolutin act as agonists or antagonists at 2c receptors and whether 2c receptors contribute to the small effect of adrenolutin on SO remain to be determined. Currently, there is no evidence that activation or blockade of 2c receptors is psychotomimetic. DA neurons can also be excited by several other classes of psychotomimetic drugs acting through opioid receptors, glutamate NMDA receptors, or cannabinoid CB1 receptors. The lack of an excitatory effect of adrenochrome suggests that the compound is unlikely to have a signicant effect on these receptors. Binding assays conrmed that all 4 catecholamine metabolites had no signicant afnity for opioid , , and receptors. They also showed no signicant binding to the 5-HT transporter and 5-HT1A,B,D,E, 2A,B,C, 3, 5A, 6, 7, adrenergic 1A,B,D, 2A,B, 1, 2, 3, histamine H14, muscarinic M15, GABAA, Sigma 1, 2, and benzodiazepine receptors. Thus, despite efforts to determine a basis for the reported psychotomimetic effect of adrenochrome, the present study failed to nd any evidence that would support those claims. The afnity of adrenochrome, adrenolutin and DHICA for the 2C receptor was unexpected and the signicance of this nding remains to be determined. References
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Jasper, J.R., Lesnick, J.D., Chang, L.K., Yamanishi, S.S., Chang, T.K., Hsu, S.A., Daunt, D.A., Bonhaus, D.W., Eglen, R.M., 1998. Ligand efcacy and potency at recombinant alpha2 adrenergic receptors: agonist-mediated [35S]GTPgammaS binding. Biochem. Pharmacol. 55 (7), 10351043. Mills, J.A., 2010. Hallucinogens as hard science: the adrenochrome hypothesis for the biogenesis of schizophrenia. Hist. Psychol. 13 (2), 178195. Rosin, D.L., Talley, E.M., Lee, A., Stornetta, R.L., Gaylinn, B.D., Guyenet, P.G., Lynch, K.R., 1996. Distribution of alpha 2C-adrenergic receptor-like immunoreactivity in the rat central nervous system. J. Comp. Neurol. 372 (1), 135165. Schwarz, B.E., Sem-Jacobsen, C.W., Petersen, M.C., 1956. Effects of mescaline, LSD-25, and adrenochrome on depth electrograms in man. AMA Arch. Neurol. Psychiatry 75 (6), 579587. Shi, W.X., Pun, C.L., Zhang, X.X., Jones, M.D., Bunney, B.S., 2000. Dual effects of Damphetamine on dopamine neurons mediated by dopamine and nondopamine receptors. J. Neurosci. 20 (9), 35043511. Shi, W.X., Pun, C.L., Zhou, Y., 2004. Psychostimulants induce low-frequency oscillations in the ring activity of dopamine neurons. Neuropsychopharmacology 29 (12), 21602167. Zhou, Y., Bunney, B.S., Shi, W.X., 2006. Differential effects of cocaine on ring rate and pattern of dopamine neurons: role of alpha1 receptors and comparison with L-dopa and apomorphine. J. Pharmacol. Exp. Ther. 317 (1), 196201.

John L. Krstenansky Department of Pharmaceutical Sciences, Loma Linda University Schools of Pharmacy and Medicine, Loma Linda, CA 92350, USA Department of Basic Sciences, Loma Linda University Schools of Pharmacy and Medicine, Loma Linda, CA 92350, USA E-mail address: jkrstenansky@gmail.com. Dan Xu Department of Pharmaceutical Sciences, Loma Linda University Schools of Pharmacy and Medicine, Loma Linda, CA 92350, USA Department of Geriatric Neurology, Chinese PLA General Hospital, Beijing, China E-mail address: dxu@llu.edu. Rolf Leitzke April Saldivar Ruzanna Gevorkian Department of Pharmaceutical Sciences, Loma Linda University Schools of Pharmacy and Medicine, Loma Linda, CA 92350, USA E-mail addresses: rleitzke@llu.edu (R. Leitzke), asaldivar@llu.edu (A. Saldivar), rgevorkian@gmail.com (R. Gevorkian). Wei-Xing Shi Department of Pharmaceutical Sciences, Loma Linda University Schools of Pharmacy and Medicine, Loma Linda, CA 92350, USA Department of Basic Sciences, Loma Linda University Schools of Pharmacy and Medicine, Loma Linda, CA 92350, USA Corresponding author at: Dept of Pharmaceutical Sci., LLU School of Pharmacy, 11262 Campus Street, West Hall, Loma Linda, CA 92350, USA. Tel.: + 1 909 558 7008; fax: +1 909 558 0446. E-mail address: wshi@llu.edu. 14 July 2011