Effect of a pyridinium metabolite derived from haloperidol on the activities of striatal tyrosine hydroxylase in freely moving rats

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Neuroscience Letters 214 (1996) 183-186

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Effect of a pyridinium metabolite derived from haloperidol on the activities of striatal tyrosine hydroxylase in freely moving rats Kazuo Igarashi a'*, Kazuo Matsubara b, Fumiyo Kasuya a, Miyoshi Fukui a, Tomoko Idzu b, Neal Castagnoli, Jr. c aFaculty of Pi~rmaceutical Sciences, Kobegakuin University, 518 Arise, Ikawadani-eho, Nishi-ku, Kobe 651-21, Japan bDepartment of Legal Medicine, Shimane Medical University, lzumo 693, Japan CDepartment of Chemistry, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061-0212, USA Received 3 June 1996; revised version received 17 July 1996; accepted 17 July 1996

Abstract

The effects of a pyridirdum metabolite (I-IPP+) derived from haloperidol (HP) on in vivo tyrosine hydroxylation was evaluated in freely moving rats. As an index of the in vivo activity of tyrosine hydroxylase (TH), the rat striatum was peffused with NSD-1015, and extracellular 3,4-dihydroxyphenylalanine (DOPA) levels were measured. HPP + (1 raM) gradually reduced tyrosine hydroxylation to 30% of the basal level, alflaough the effect was less potent than 1-methyl-4-phenylpyridinium ion (MPP+). On the contrary, HPP + at a 0.1 mM dose decreased in 5-hydroxyindoleacetic acid (5-HIAA) level, but did not affect dopamine metabolites. The present study revealed that HPP + irreversible inhibited in vivo tyrosine hydroxylation by the same manner of MPP +. However, the neurotoxie effects of I-IPP+ in vivo would be selective for serotonergic over dopaminergic neurons, which distinguishes the toxic profile of this compound compared to that of MPP +.

Keywords: Pyridinium metabolite; Haloperidol; 1-Methyl-4-phenylpyridinium ion; Tyrosine hydroxylase; Microdialysis

The neurotoxin 1-methyl-4-phenyl-l,2,3,6-tetrahydropyridine (MPTP) damages the nigrostriatal dopaminerigic (DA) system in animal.s and humans [11]. The neurotoxic effects of MPTP are caused by its active metabolite, 1methyl-4-phenylpyridinium ion (MPP+), via monamine oxidase B [4,12]. Several analogs of MPTP, that are metabolized to the corresponding pyridinium derivatives, are known to display MPTP-type neurotoxicity in animals [3,13,16,21,28-30]. The antipsychotic agent haloperidol (HP) (Fig. 1) is a piperidinol derivative which undergoes chemical [24] and/or enzymatic dehydration [5,6] to produce the 1,2,3,6-tetrahydropyridine derivative (HPTP) (Fig. 1), which bears structural features similar to those of MPTP. These findings have prompted us to consider that the side effects as,sociated with chronic HP use [1], such as the tardive dyskinesias, might be mediated by the 4-(4-chlorophenyl)- 1-[,4-(4-fluorophenyl)-4-oxobutyl]pyr* Corresponding author. Tel.:+81 78 9741551, ext. 2488; fax: +81 78 9745689.

idinium ion (HPP +) (Fig. 1), a structural analog of MPP +. The results from earlier studies documented the oxidative biotransformation of I-1P to HPP + in both rodents [25] and humans [9,26]. More recently, we have reported that HPP + levels in brain tissues of rats increased gradually along with the HP administration [10]. The results form intracerebral microdialysis [22] and neuronal cell culture [2] studies have shown that HPP + has toxic effects on dopaminergic and serotonergic neurons resembling those of MPP +. Moreover, HPP + is a more potent inhibitor of mitochondrial respiration than MPP + in vitro. MPTP and MPP + are known to affect the activity of tyrosine hydroxylase (TH), the rate-limiting enzyme of dopamine synthesis. Repeated administration of MPTP to mice reduce both TH activity and TH protein in the striatum [17]. MPTP and MPP + reduce tyrosine hydroxylation in rat striatal tissues [8,20]. The recent study using microdialysis have revealed that MPP + reduced in vivo doparnine synthesis possibly through the inhibition of phosphorylation of TH [14]. On the other hand, the effect

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K. Igarashi et al. / Neuroscience Letters 214 (1996) 183-186

continuously perfused for 120 min to measure the basal in vivo tyrosine hydroxylation. Subsequently, MPP + (0.1 or 1 mM) or HPP ÷ (0.1 or 1 mM) in Ringer's solution containing 10 /zM NSD-1015 was perfused for 60 min followed by sequential perfusion with Ringer's solution containing 10/zM NSD-1015 for 180 min. The dialysate was collected every 20 min during the perfusion. Dialysate concentrations of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), DOPA, homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) were measured using high performance liquid chromatography (HPLC). The analytes were separated on a reverse phase C18 column (Eicompack MA-5ODS, 150 × 4.6 mm, EICOM, Japan), and detected electrochemically with a glassy carbone working electrode, set at +750 mV versus Ag/AgC1 (EICOM, Japan). The mobile phase, 0.1 M citric acid/ sodium acetate buffer (pH 2.8) containing 1.2 mM 1-octanesulfonic acid, 10 mM EDTA and 15% methanol (v/v), was delivered at a flow rate of 0.8 ml/min by a Shimadzu LC-10A HPLC pump. Statistical analysis was performed using two-way analysis of variance (ANOVA) with repeated measures on one factor; the post hoc Dunnett ttest was used to determine statistical significance between drug treatments. In the present study, we used a microdialysis technique to examine the acute effect of HPP ÷ on the in vivo tyrosine hydroxylation. Fig. 2 shows the time-response curves for the effects of HPP ÷ and MPP ÷ on extracellular levels of striatal DOPA in the presence of 10 #M NSD-1015, an inhibitor of aromatic L-amino acid decarboxylase. Before addition of NSD-1015 into the perfusion medium, DOPA was not detectable in the striatal dialysate. During perfusion of NSD-1015, DOPA levels increased steadily to reach a stable level within 100-120 min (363.1 + 22.4 fmol/min, mean _+ SEM, which was equivalent to in vivo TH activity). MPP ÷ at all experimental doses gradually

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of HPP ÷ on the TH activity has not been studied yet. In an effort to characterize further its neurotoxic potential, we undertook the experiment to asseSs the acute effect of HPP ÷ on in vivo tyrosine hydroxylation using a recently developed microdialysis technique [7,27]. Anesthetized (50 mg/kg i.p., sodium pentobarbital) male Wistar rats (250-300 g, SLC) were stereotaxically implanted with 22-gauge cannulae in the left striata at AP +0.48 mm, L +2.8-2.9 mm from the bregma, and -3.5 mm from the skull, according to the stereotaxic atlas of Paxinos and Watson [19]. Dummy probes were then placed inside the cannulae. The rats were housed in plastic cages (35 x 35 x 40 cm) with free access to food and water, and a 20 h recovery period was allotted. The microdialysis probes with dialysis area of 3 mm length were of the Ishaped type reported previously [18]. The dialysis tube (0.2 mm i.d., 0.31 mm o.d.) was prepared from a polyacrylonitrile/sodium methylsulfonate membrane (Hospal, Bologna) with a molecular weight cut-off of 1100 Da. After insertion through the guide cannulae, the probe was connected to a microinfusion pump and perfused with Ringer's solution at a flow rate of 2 #l/min for 180 min. Then, the perfusion medium was replaced with Ringer's solution containing 10 #M m-hydroxybenzylhydrazine (NSD-1015) and the striatal specimen was MPP m +m or H P P + 150

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K. Igarashi et al. / Neuroscience Letters 214 (1996) 183-186

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Fig. 3. Effects of HPP÷ and MPP÷ on 5-HIAA level in the presence of 10 #M NSD-1015. After steady-state production of 5-HIAA was established, varying concentration of HPP÷ (closed triangle and square) and MPP+ (open Iriangle and square) were added to the perfusion medium for 60 min as indicated by a bar. Data are mean + SEM values expressed as percentages of the steady-state 5-HIAAproduction. Number of animals used are shown in parentheses. reduced D O P A levels !in the dialysate (P < 0.01). Perfusion with HPP ÷ at 1 mM dose also displayed a similar effect (P < 0.01). The perfusion of either HPP ÷ (1 mM) or MPP ÷ (0.1 and 1 mM) decreased D O P A level in the dialysate irreversibly, at least, during the experiment. However, HPP ÷ at 0.l m M did not cause a decrease in extraceUular DOPA. "I~ese results indicate that the two compounds have a similar inhibitory mechanism on TH activity, although HPP ÷ is less potent than MPP ÷. The inhibition of tyrosine hydroxylation by MPP ÷ is probably not due to direct inhibition of TH but to restraint of the enzyme activation through continuous inhibition of the enzyme phosphorylation [14]. Inhibitory effect of HPP ÷ on TH activity is demonstrated first in the present study. However, a lower dose (0.1 mM) of HPP ÷ did not affect in vivo tyrosine hydroxylation. Also, total DA content released by perfusion with HPP ÷ was 4 - 5 fold less than that released by MPP ÷ (not shown). These results might be explained by lower affinity for the active transporter of dopamine neurons cornpared with MPP ÷. The concentrations of HPP ÷ in the striatum has been reported to be less than 50 pmol/g and 1 mnol/stdatum (ca. 10 nmol/g) after 0.1 and 1 m M perfusion for 2 h, respectively [21]. MPP ÷ levels has been ca. 6 and 94 nmol/g after 2 h perfusions with 0.1 and 1 m M solutions, respectively [21]. On the other hand, daily interaperitonial administration of HP leads to the gradual accumulation of HPP ÷ in the brain; the level reaches to ca. 150 pmol/g striatum only after daily 10 mg/kg HP for three days in rat [10]. However, it is not clear whether chronic treatment with HP [15,23,31] to animals cause structural damage of neuronal cells. Moreover, the side effects of HP, such as tradive dyskinesia, are usually occurred after months or years of treatment in human [1] and therefore HPP ÷ concentration in the striatum after long-term HP use in patients might be high enough to inhibit ~[7-Iactivity in the striatum, although

the actual brain level has not been reported in these patients. We also determined the acute effects of HPP ÷ and MPP + on the extracellular levels of DOPAC, H V A and 5-HIAA during perfusion of NSD-1015. Perfusion with HPP + at 0.1 and 1 mM doses significantly decreased 5H I A A levels in the dialysis (P < 0.01; Fig. 3), but did not affect DOPAC nor H V A level compared to those of control (not shown). On the contrary, MPP ÷ at 0.1 m M solution did not change 5-HIAA level (Fig. 3), but decreased DOPAC and H V A levels (P < 0.01, data not shown). These data may confirm the previous data that the toxicity of HPP + is selective for serotonergic over dopaminergic neurons, which distinguishes the toxic profile of this compound compared to that of MPP + [21]. In conclusion, the present study revealed that HPP ÷ inhibited in vivo tyrosine hydroxylation by the same manner of MPP +. However, the neurotoxic effects of HPP + in vivo would be different from those of MPP +, especially in the selectivity for dopaminergic neurons. The animal experiments were done in accordance with the guidelines for care and use of laboratory animals by the Committee of Shimane Medical University. A part of this work was supported by a Grant-in-Aid for Scientific Research in Japan. [1] Baidessarini, R.J., Drugs and the treatment of psychiatric disorders. In A.G. Gilman, T.W. Rail, A.S. Nies and P. Taylor (Eds.), Goodman and Gilman's The PharmacologicalBasis of Therapeutics, 8th edn., Pergamon Press, New York, 1991, pp. 400. [2] Bloomquist, J., King, E., Wright, A., Mytilineoli, C., Klmura, K., Castagnoli, K. and Castagnoli, N. Jr., 1-Methyl-4-phenylpyfidinium-like neurotoxicity of a pyridinium metabolite derived from haioperidol: cell culture and neurotransmitter uptake studies, J. Pharmacol. Exp. Ther., 270 (1994) 822-833. [3] Booth, R.G., Castagnoli, N. Jr. and RoUema, H., Intracerebral

186

[4]

[5]

[6]

[7]

[8]

[9]

[10]

[11]

[12]

[13]

[14]

[15]

[16]

[17]

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microdialysis neurotoxicity studies of quinoline and isoquinoline derivatives related to MPTP/MPP ÷, Neurosci. Lett., 100 (1989) 306-312. Chiba, K., Trevor, A. and Castagnoli, N. Jr., Metabolism of the neurotoxic tertiary amine MPTP by brain monoamine oxidase, Biochem. Biophys. Res. Commun., 120 (1985) 574-578. Fang, J. and Gorrod, J.W., Dehydration is the first step in the bioactivation of haloperidol to its pyridinium metabolites, Toxicol. Lett., 59 (1991) 117-123. Fang, J., Yu, P.H., Gorrod, J.W. and Boulton, A.A., Inhibition of monoamine oxidases by haloperidol and its metabolites: pharmacological implications for the chemotherapy of schizophrenia, Psychopharmacology, 118 (1995) 206-212. Hashiguti, H., Nakahara, D., Mamyama, W., Naoi, M. and Ikeda, T., Simultaneous determination of in vivo hydroxylation of tyrosine and tryptophan in rat striatum by microdialysis-HPLC: relationship between dopamine and serotonin biosynthesis, J. Neural Transm., 93 (1993) 213-223. Hirata, Y. and Nagatsu, T., Inhibition of tyrosine hydroxylation in tissue of the rat striatum by 1-methyl-4-phenyl-l,2,3,6tetrahydropyridine, Brain Res., 337 (1985) 193-196. Igarashi, K., Kasuya, F., Fukui, M., Abe, T. and Castagnoli, N. Jr., Simulataneous determination of haloperiodol and its neurotoxic metabolite in plasma and brain tissue from schizophrenic patients treated with haloperiodol using HPLC and solid-phase extraction, Jpn. J. Forensic Toxicol., 13 (1995) 31-38. Igarashi, K., Kasuya, F., Fukui, M., Usuki, E. and Castagnoli, N. Jr., Studies on the metabolism of haloperidol (HP): the role of CYP3A in the production of the neurotoxic pyridinium metabolite HPP ÷ found in rat brain following i.p. administration of HP, Life Sci., 57 (1995) 2439-2446. Langston, W.J., Mechanisms underlying neuronal degeneration in Parkinson's Disease: an experimental and theoretical treatise, Movement Disorders (Suppl.), 1,4,S (1989) 15-125. Markey, S.P., Johannessen, J.N., Chiurh, C.C. and Burns, R.S., Intraneuronal generation of a pyridinium metabolite may cause drug-induced Parkinsonism, Nature, 311 (1984) 464-467. Matsubara, K., Neafsey, E.J. and Collins, M.A., Novel S-adenosylmethionine-dependent indole-N-methylation of B-carbolines in brain particulate fractions, J. Neurochem., 59 (1992) 511-518. Matsubara, K., Idzu, T., Kobayashi, Y., Nakahara, D., Maruyama, W., Kobayashi, S., Kimura, K. and Naoi, M., N-Methyl-4-phenylpyridinium and an endogenously formed analog, N-methylated Bcarbolinium, inhibit striatal tyrosine hydroxylation in freely moving rats, Neurosci. Lett., 199 (1995) 199-202. Mereu, G., Lilliu, V., Vargiu, P., Muntoni, A.L., Diana, M. and Gessa, G.L., Failure of chronic haloperidol to induce depolarization inactivation of dopamine neurons in unanesthetized rats, Eur. J. Pharmacol., 264 (1994) 449-453. Michel, P.P., Dandapani, B.P., Sanchez-Ramos, J., Efange, S.J., Pressman, B.C. and Hefti, F., Toxic effects of potential environmental neurotoxins related to 1-methy174-phenylpyddininm on cultured rat dopaminergic neurons, J. Pharmacol. Exp. Ther., 248 (1989) 842-850. Mogi, M., Harada, M., Kojima, K., Kiuchi, K., Nagatsu, L. and Nagatsu, T., Effects of repeated systemic administration of 1methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on striatal tyrosine hydroxylase activity in vitro and tyrosine hydroxylase content, Neurosci. Lett., 80 (1987) 213-218.

[18] Nakahara, D., Ozaki, N. and Nagatsu, T., A removed brain microdialysis probe units for in vivo monitoring of neurochemical activity, Biogenic Amines, 6 (1989) 559-564. [19] Paxinos, G. and Watson, C., The Rat Brain in Stereotaxic Coordinates, 2nd edn., Academic Press, New York, 1986. [20] Pileblad, E., Fomstedt, B., Clark, D. and Carlsson, A., Acute effects of 1-methyl-4-phenyl-l,2,3,6-tetrahydropyridine on dopamine metabolism in mouse and rat striatum, J. Pharm. Pharmacol., 37 (1985) 700-712. [21] Rollema, H., Johnson, E.A., Booth, R.G., Caidera, P., Lampen, P., Youngster, S.K., Trevor, A., Naiman, N. and Castagnoli, N. Jr., In vivo intracerebral microdialysis studies in rats of MPP ÷ analogs and related charged species, J. Meat. Chem., 33 (1990) 2221-2230. [22] Rollema, H., Skolnik, M., D'Engelbronner, J., Igarashi, K., Usuki, E. and Castagnoli, N. Jr., MPP ÷-like neurotoxicity of a pyridinium metabolite derived from haloperidol: in vivo microdyalysis and in vitro mitochondrial studies, J. Pharmacol. Exp. Ther., 268 (1994) 380-387. [23] See, R.E., Chapman, M.A. and Meshul, C.K., Comparison of chronic intermittent haloperidol and raclopride effects on striatal dopamine release and synaptic ultrastructural in rats, Synapse, 12 (1992) 147-154. [24] Subramanyam, B., Rollema, H., Woolf, T. and Castagnoli, N. Jr., Identification of a potentially neurotoxic pyridinium metabolite of baloperidol in rats, Biochem. Biophys. Res. Commun., 166 (1990) 238-244. [25] Subramanyam, B., Woolf, T. and Castagnoli, N. Jr., Studies on the in vitro conversion of haloperidol to a potentially neurotoxic pyridinium metabolites, Chem. Res. Toxicol., 4 (1991) 123-128. [26] Subramanyam, B., Pond, S.M., Eyles, D.W., Whiteford, H.A., Fouda, H.G. and Castagnoli, N. Jr., Identification of a potentially neurotoxic pyridmium metabolite in the urine of schizophrenic patients treated with haloperidol, Biochem. Biophys. Res. Commun., 181 (1991)573-578. [27] Westerink, B.H.C., De Vries, J.B. and Duran, R., Use of microdialysis for monitoring tyrosine hydroxylase activity in the brain of conscious rats, J. Neurochem., 54 (1990) 381-387. [28] Youngster, S.K., SonsaUa, P.K. and Heikkila, R.E., Evaluation of the biological activity of several analogs of the dopaminergic neurotoxin 1-methyl-4-phenyl- 1,2,3,6-tetrahydropyfidine, J. Neurochem., 48 (1987) 929-934. [29] Youngster, S.K., Nicldas, W.J. and Heikkla, R.E., Structure-activity study of the mechanism of 1-methyl-4-phenyi-l,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity. II. Evaluation of the biological activity of the pyridinium metabolites formed from the monoamine oxidase-catalyzed oxidation of MPTP analogs, J. Pharmacol. Exp. Ther., 248 (1989) 828-835. [30] Youngster, S.K., Sonsalla, P.K., Sieber, B.E. and Heikldla, R.E., Structure-activity study of the mechanism of 1-methyl-4-phenyl1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity. I. Evaluation of the biological activity of MPTP analogs, J. Pharmacol. Exp. Ther., 248 (1989) 820-828. [31] Zhang, W., Tilson, M.K., Srachowisk and Hong, J.S., Repeated haloperidol administration changes basal release of striatal dopamine and subsequent response to haloperidol challenge, Brain Res., 484 (1989) 389-395.