Shin-ichi Iwata

Antinociceptive mechanism of l-DOPA

&NA; The mechanism of l‐DOPA for antinociception was investigated. Nociceptive behaviors in mice after an intrathecal (i.t.) administration of substance P were evaluated. l‐DOPA (i.t.) dose‐dependently attenuated the substance P‐induced nociceptive behaviors. Co‐administration of benserazide (i.t.), a DOPA decarboxylase inhibitor, abolished the antinociceptive effect of l‐DOPA. The l‐DOPA‐induced antinociception was antagonized by sulpiride, a D2 blocker, but not by SCH 23390, a D1 blocker. These results suggest that l‐DOPA relieves pain after conversion to dopamine, with the dopamine sedating pain transmission by way of the dopamine D2 receptor.

Effect of selegiline on dopamine concentration in the striatum of a primate

Monoamine oxidase (MAO) has two subtypes, A and B, that have different distributions between the rodent and the human. In the striatum, dopamine (DA) of the rat seems to be metabolized by MAO A, and DA of the human is largely deaminated by MAO B. MAO in the striatum of common marmosets is also type B. Using in vivo microdialysis, we investigated the pharmacological activity of selegiline, a selective irreversible inhibitor of MAO B, in the striatum of marmosets. Intraperitoneal co-administration of selegiline (1 mg kg-1, i.p.) with levodopa/carbidopa (10/2.5 mg kg-1, i.p.) did not significantly increase extracellular concentration of DA in the striatum of common marmosets compared with control animals receiving levodopa/carbidopa alone. Daily pretreatment with 0.1 mg kg-1 (i.p.) selegiline for two weeks, however, dramatically increased extracellular concentration of DA to about seven times that of control animals treated with levodopa/carbidopa alone in marmosets. Such an increase in extracellular concentrations of DA could not be observed in a similar study with Wistar rats. This study showed that chronic administration of a small dose of selegiline caused a marked increase in extracellular DA concentration in the striatum of primates, but not in the rodents.

The metabolic rate and vulnerability of dopaminergic neurons, and adenosine dynamics in the cerebral cortex, nucleus accumbens, caudate nucleus, and putamen of the common marmoset.

Abstract The pathophysiology of the striatum and cerebral cortex were studied from the pharmacological aspect.Investigation of the dopamine content in the cerebral cortex revealed that the premotor and motor area showed the highest level (61±6.2 ng/g). Intravenous injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) at a dose of 10 mg/kg reduced the dopamine content in the caudate nucleus and putamen to 2–3% of the control level in common marmosets, while it fell to 60% in the nucleus accumbens. There was no alteration of the dopamine content in the cerebral cortex. Immunohistochemical staining for tyrosine hydroxylase in the midbrains of MPTP-treated marmosets showed almost complete disappearance of dopaminergic cells from the substantia nigra and good preservation of cells in the ventrotegmental area. Dopaminergic cells projecting to the caudate/putamen, nucleus accumbens, and cerebral cortex showed marked, moderate, and no vulnerability to MPTP, respectively.After systemic administration of MPTP, dopaminergic neurons projecting to the caudate nucleus and putamen were damaged equally. However, the compensatory increase of dopamine turnover was more prominent in the putamen than in the caudate nucleus. Thus, nigroputaminal dopaminergic neurons may have a higher level of activity than neuron in the caudate. The neural connections and functions of the caudate nucleus and putamen have already been differentiated anatomically or physiologically. This compensatory increase of the dopamine turnover rate is another aspect of functional differences between the caudate nucleus and putamen.Investigation of the dopamine content in the heat, body, and tail of the caudate nucleus showed no differences in the concentration of dopamine. However, a study of the metabolic rate of dopamine using α-methyl-p-tyrosine, a tyrosine hydoxylase inhibitor, showed higher metabolism of dopamine in the head of the caudate nucleus in common marmosets. Thus, dopaminergic neurons projecting to the caudate nucleus may show topographical differences in their firing rates.A microdialysis study indicated an increase in the metabolism of adenosine in the striatum of MPTP-treated animals. Cholinergic neurons are interneurons and are one of the main sources of adenosine in the striatum. Dopaminergic input from the substantia nigra acting on cholinergic neurons was decreased following MPTP treatment. The increase of adenosine metabolism suggested that cholinergic neurons in the striatum receive inhibitory inputs from nigrostriatal dopaminergic neurons.

Effects of Acute or Prolonged Administration of Cabergoline on Parkinsonism Induced by MPTP in Common Marmosets

The effects of a single treatment or chronic administration of cabergoline (1-[(6-allylergolin-8beta-yl)carbonyl]-1-[3-(dimethylamino)p ropyl]-3-ethyl-urea), a potent, long-lasting dopamine receptor agonist, on parkinsonism induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in common marmosets were studied. The administration of 0.2 mg/kg or a higer dose of cabergoline began to reverse parkinsonism-like symptoms 60 min after a subcutaneous injection, and showed steady and constant effects throughout the observation period. For prolonged administration, 0.2 mg/kg cabergoline was injected daily for 22 consecutive days. Locomotor activity in MPTP-treated animals increased until it reached its peak on the third day, then it gradually decreased. Akinesia scores, rating the quality of movements, were also improved, and the improvement was sustained up to the last day of chronic administration. None of the animals developed abnormal behaviors after either acute or chronic administration. These results suggest that cabergoline has long-acting effects in the marmoset model of parkinsonism, and that it will be a useful agent for the treatment of Parkinsons disease, particularly in cases with fluctuating motor disabilities.

Increased dopamine turnover in the putamen after MPTP treatment in common marmosets

The differences in dopamine turnover rate between the putamen and the caudate nucleus in the striatum lesioned by a neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) were studied in the common marmoset, a small New World monkey. Systemic administration of MPTP damaged equally and dose-dependently nigrostriatal dopaminergic neurons projecting both to the caudate nucleus and the putamen. The compensatory increase of dopamine turnover, however, occurred more prominently in the putamen than in the caudate. The neural connection and function of the caudate nucleus and the putamen have been differentiated anatomically or physiologically. The compensatory increase of dopamine turnover rate is another different aspect of functions between the caudate nucleus and the putamen. Dopaminergic neurons projecting to the putamen showed more prominent cell loss than those projecting to the caudate in Parkinsons disease or related disorders. The selective augmented turnover rate of lesioned dopaminergic neurons might be, at least partly, involved with selective degeneration of nigrostriatal neurons projecting to the putamen.

Delayed L-DOPA-induced hyperalgesia.

Previously we reported on L-DOPAs antinociceptive effect on substance P-induced nociceptive behaviors in mice [Shimizu T, Iwata S, Morioka H, Masuyama T, Fukuda T, Nomoto M. Antinociceptive mechanism of L-DOPA. Pain 2004;110;246-9.]. Since significant hyperalgesia was noted following antinociception, our study was designed to investigate the mechanism of this hyperalgesia. Nociceptive behaviors were enhanced 2 h after L-DOPA administration. L-DOPA induced hyperalgesia occurred after conversion to dopamine because co-administration of benserazide, a DOPA decarboxylase inhibitor, completely abolished the L-DOPA-induced hyperalgesia. The D2 receptor agonist, quinpirole, depressed these behaviors entirely, while the D1 antagonist, SCH23390, inhibited the enhancement of these behaviors by L-DOPA. The D2 receptor antagonist, sulpiride, which induced hyperalgesia of the substance P-induced behaviors in naive mice, did not have any effects on L-DOPA-induced hyperalgesia. Spinal cord dopamine content increased rapidly after L-DOPA administration, exhibiting levels 100 times greater than baseline, and then returned to control after 1 h. These results suggested that the dopaminergic inhibitory system for pain sensation was temporarily impaired by excess amounts of exogenous dopamine that were derived from L-DOPA and both D1 and D2 receptors were involved in L-DOPA-induced hyperalgesia.

Effects of β-adrenergic blockers on drug-induced tremors

Abstract We studied the effect of various kinds of β-adrenergic blockers on oxotremorine-, harmaline- and thyrotropin-releasing hormone (TRH)-induced tremors in mice. To investigate what property of β-blockers plays the main role in suppressing tremor, we employed five β-blockers (propranolol, atenolol, butoxamine, pindolol, and arotinolol). All drugs suppressed oxotremorine-induced tremors but none reduced harmaline-induced tremors. Even though TRH-induced tremors were decreased significantly only by propanolol and high doses of arotinolol, all drugs had a tendency to reduce the tremor. We concluded that neuropharmacological mechanisms underlying to harmaline-induced tremors were different from those of TRH- and oxotremorine-induced tremors and that features of β-blockers ( β 1 or β 2 -selectivity, intrinsic sympathomimetic activity, and membrane stabilizing activity) did not primarily contribute to the suppression of tremors.

TH protein and mRNA in nigrostriatal dopaminergic neurons are down-regulated by continuous but not intermittent apomorphine.

Levels of tyrosine hydroxylase (TH) and TH mRNA were measured after administration of dopamine agonists for a long period of time to elucidate the long-term feedback inhibition of dopamine synthesis in nigrostriatal dopaminergic neurons. Continuous infusion, which desensitized presynaptic dopamine receptors, but not repeated administration, down-regulated TH and TH mRNA levels. This suggests levels of TH protein and mRNA are only feedback inhibited by the continuous stimulation of postsynaptic dopamine receptors.

Effects of repeated testing on the incidence of haloperidol-induced catalepsy in mice

Effects of repeated testing on the incidence of haloperidol-induced catalepsy were investigated in mice. The incidence of catalepsy, evaluated with the forelimbs or hindlimbs placed on a standard horizontal bar, increased in three successive tests in mice injected with haloperidol. Catalepsy was not provoked by repeated testing in animals with saline. In a subsequent study, mice were examined for catalepsy in the forelimbs in the first two trials and then in the hindlimbs. In this procedure, the incidence of catalepsy did not increase with repeated testing. These results suggest that repeated testing increases the incidence of haloperidol-induced catalepsy but does not influence the cataleptogenic potency of the drug.

Upregulation of postsynaptic dopamine receptors in the striatum does not influence haloperidol-induced catalepsy in mice.

The incidence of haloperidol-induced catalepsy was investigated in mice whose postsynaptic dopamine (DA) receptors in the striatum had been upregulated by denervation with 6-hydroxydopamine (6-OHDA). In nonupregulated mice, which were injected with 6-OHDA 4 days before, DA in the striatum fell to 21% of the level found in vehicle-injected mice but [3H]spiperone binding to the membrane of the striatum did not increase. In upregulated mice, which were injected with 6-OHDA 28 days before, DA was at 24% and [3H]spiperone binding increased by 15%. The ED50 values (with 95% confidence limits) for haloperidol-induced catalepsy in nonupregulated mice and that in upregulated mice was 0.40 mg/kg (0.25-0.65 mg/kg) and 0.29 mg/kg (0.16-0.51 mg/kg), respectively. There was no significant difference in the incidence of catalepsy between the two groups of mice. This suggests that the intensity of catalepsy produced by the DA receptor blockade may be unaltered even when the density of receptors increases.