Wacław Kolasiewicz

MESOLIMBIC NMDA RECEPTORS ARE IMPLICATED IN THE EXPRESSION OF CONDITIONED MORPHINE REWARD

Abstract Systemic administration of a variety of N-methyl-d-aspartate (NMDA) receptor antagonists inhibits morphine’s rewarding properties in the conditioned place preference test. In this study, we investigated the anatomical loci implicated in the inhibition of expression of morphine’s reward by bilateral microinjections of a selective NMDA antagonist into the mesolimbic areas, including ventral tegmental area and nucleus accumbens. During conditioning, injections of 1 mg/kg morphine were associated with placing rats in one chamber of the place preference box; the exposures to the other chamber were associated with placebo administration. On the test day, drug-free control subjects demonstrated a marked preference for the morphine-associated chamber. Systemic administration of 5 mg/kg and 10 mg/kg of the competitive NMDA antagonist, NPC 17742 (2R,4R,5S-2-amino-4,5-(1,2-cyclohexyl)-7-phosphonoheptanoic acid), significantly reduced the expression of morphine-induced conditioned place preference; the dose of 10 mg/kg produced also an inhibition of locomotor activity. Similar attenuation of the expression of morphine-induced conditioned place preference was observed in rats receiving 15.6 and 62.5 ng/0.5 μl side of NPC 17742 injected bilaterally into the nucleus accumbens and ventral tegmental area. While the higher intra-accumbal dose of NPC 17742 produced behavioral stimulation, intra-tegmental injection did not affect locomotor activity. These findings suggest that activation of NMDA receptors in the nucleus accumbens and ventral tegmental area is necessary for the elicitation of approach by environments previously associated with morphine’s rewarding action.

Substantia nigra as a station that not only transmits, but also transforms, incoming signals for its behavioural expression: Striatal dopamine and GABA-mediated responses of pars reticulata neurons

Abstract In order to investigate whether striatal dopaminergic mechanisms are involved in the behavioural expression of the GABAergic mechanisms within the pars reticulata of the substantia nigra, apomorphine or haloperidol were bilaterally administered into the caudate nucleus of cats pretreated with a unilateral injection of picrotoxin or muscimol into the nigral pars reticulata. Although the doses selected for the intracaudate injections have been shown to be maximally effective in affecting the behavioural expression of the caudate function, the pharmacological treatment of the caudate nucleus did not produce any significant change in the behaviour elicited from the nigra; neither the picrotoxin-induced asymmetric posturing, asymmetric circling, freezing and hind leg disorders nor the muscimol-induced asymmetric posturing, asymmetric spinning and stereotyped licking were significantly affected. The latter behaviour was absent in animals with a partial or total destruction of the nigral pars reticulata. As the behavioural expression of the nigral pars reticulata differed completely from the asymmetric head twisting known to be characteristic for the caudate nucleus, it is suggested that the behavioural expression of the caudate nucleus requires a main output station elsewhere in the brain. Furthermore, the present results demonstrate that the nigral pars reticulata does not form part and parcel of a feedback system that simply transmits incoming signals from the caudate nucleus towards the pars compacta, i.e. the origin of the dopaminergic, nigrostriatal system. Finally, the present study demonstrates that the dopaminergic activity within the caudate nucleus may only modify, but certainly not determine, the behavioural expression of the nigral pars reticulata. It is concluded that the nigral pars reticulata not only transmits, but also transforms its incoming signals.

The effects of muscimol and picrotoxin injections into the cat substantia nigra.

SummaryThe behaviour of cats after unilateral injections of muscimol, picrotoxin and bicuculline into the posterior and lateral parts of the substantia nigra was observed. The antagonism between muscimol and picrotoxin was limited to some effects of the drugs: (1) muscimol-induced sniffing, licking and enhanced locomotor activity was attenuated by picrotoxin in contrast to the muscimol-induced contralateral turning which was even enhanced by picrotoxin; (2) picrotoxin-induced balance disorders as well as characteristic motor disturbances in the hind legs remained unaffected following an additional treatment with muscimol. Furthermore, subcutaneously given apomorphine did not affect the picrotoxin-induced balance and hind leg disorders, although the apomorphine-induced stereotyped behaviour itself was enhanced and suppressed by intranigral administration of muscimol and picrotoxin respectively.The conclusion is reached that there are at least two distinct populations of GABA receptors within the substantia nigra: one group controlling or being controlled by dopaminergic neurons and one group operating fully independently of the former neurons. The similarities to experiments on rats are stressed. The mechanism underlying the muscimol-resistant picrotoxin-induced effects (balance disorders and motor disturbance of the hind legs) is discussed.

Thalamus as a relay station for catalepsy and rigidity

The aim of the study was to determine to what extent catalepsy and tonic rigidity of muscles induced by muscimol administration into the ventral thalamic nuclei disturb the motor activity of rats. This study also aimed to test whether the ventromedial thalamic nucleus (Vm) was involved in transmitting effects evoked by the systemic injection of neuroleptics or opioids. For this purpose muscimol and/or picrotoxin was injected into the ventral thalamic nuclei and the behaviour of the animals was assessed in a series of test situations. It was found that muscimol administration to the Vm disturbs not only the initiation and performance of voluntary movements but also the occurrence of avoidance when the animals life is endangered. Postural reflexes remained, however, undisturbed. Those effects seemed to be GABA- and site-specific to Vm. The haloperidol catalepsy was strongly inhibited by administration of picrotoxin to the Vm while the morphine catalepsy remained unchanged after picrotoxin. The Vm plays a crucial role in the motor behaviour and transmission of cataleptogenic effects of haloperidol, whereas similar effects produced by morphine appear to by-pass the investigated thalamic region.

Glycogen Synthase Kinase 3β and Its Phosphorylated Form (Y216) in the Paraquat-Induced Model of Parkinsonism

Parkinson’s disease is a slowly progressing disease, due to a lesion of dopaminergic neurons in the substantia nigra and a dramatic loss of dopamine in the striatum. It is now accepted that several environmental agents including the herbicide paraquat (PQ) may contribute to its pathogenesis. However, till now nothing is known about the role of glycogen synthase kinase-3β (GSK-3β) in the PQ toxicity. Therefore, the aim of this study was to examine the influence of 37-week administration of PQ in rats on the immunohistochemically measured levels of the total GSK-3β and its active, tyrosine 216 (pY216)—phosphorylated form in subcellular fractions of the midbrain with pons, as well as of the striatum. The present results revealed that the long-term PQ administration increased the levels of total and active forms of GSK-3β in the midbrain with pons, whereas decreased them in the striatum. Examination of the lesion extent showed a decrease in the number of tyrosine-immunoreactive neurons in the substantia nigra pars compacta, ventral tegmental area, and locus coeruleus, as well as lower DOPAC/dopamine ratio and noradrenaline level in the striatum in rats treated with PQ. The long-term PQ administration disturbed also motor activity of rats. Summarizing, the present data indicate that the long-term exposure of rats to PQ, a commonly used herbicide, diversely alters levels of GSK-3β in different brain structures, which may be associated with their vulnerability to its toxicity.

The role of the ventromedial thalamic nucleus in the catalepsy evoked from the substantia nigra pars reticulata in rats

Picrotoxin (25, 50 and 100 ng), injected unilaterally into the posterior part of the substantia nigra pars reticulata (SNR) of rats, evoked a dose-dependent catalepsy. The catalepsy evoked by 100 ng of picrotoxin injected into the SNR was abolished by a subsequent bilateral injection of the same drug (200 ng) into the ventromedial thalamic nuclei. It is suggested that impulses pertinent to the catalepsy evoked from the SNR are transmitted via a GABAergic pathway to the ventromedial thalamic nucleus, wherefrom they reach the striatum, as had been shown previously.

Endogenous alkaloids in man XXVI. Determination of the dopaminergic neurotoxin 1-trichloromethyl-1,2,3,4-tetrahydro-β-carboline (TaClo) in biological samples using gas chromatography with selected ion monitoring

Highly chlorinated beta-carbolines have a potential in vivo relevance to Parkinsons disease. In this paper, a gas chromatographic method for the determination of the neurotoxic 1-trichloromethyl-1,2,3,4-tetrahydro-beta-carboline (TaClo), the condensation product of tryptamine and chloral hydrate, is described. The specific and sensitive assay involves purification of the biological samples by solid-phase extraction with C18 cartridges, derivatization with heptafluorobutyric anhydride, and chromatography on a non-polar fused-silica capillary column. Detection of TaClo was achieved by the registration of characteristic mass fragments of the TaClo heptafluorobutyric amide derivative using selected ion monitoring. The method was utilized to detect and quantify TaClo in blood, urine, bile, faeces, and brain tissue of rats treated with this alkaloid-type heterocycle. Four-fold deuterium-labelled TaClo was used as an internal standard.

Effects of intrastriatal injections of atropine and methacholine on the apomorphine-induced gnawing in the rabbit

To find out the anatomical location of the target point of cholinergic-dopaminergic equilibrium, atropine (40 microng) or methacholine (10 microng) were injected through previously implanted cannulas into various places of caudate nucleus and putamen of the rabbit, and the effect of the injections on stereotype gnawing induced by subcutaneously or intravenously administered apomorphine (1--2 mg/kg) was assessed. The intensity of gnawing was measured using a special apparatus, counting each bite. Atropine inhibited the stereotype, while metacholine potentiated it. The effects were evident with the method used, but difficult to reveal with the classical method of assessing the intensity of stereotyped behavior, based on visual observation. The results suggest that the striatum is not a target point for the cholinergic component of the cholinergic-dopaminergic equilibrium in the central nervous system.

6-OHDA injections into A8–A9 dopaminergic neurons modelling early stages of Parkinson's disease increase the harmaline-induced tremor in rats

The aim of the present study was to examine the influence of a unilateral 6-hydroxydopamine (6-OHDA)-induced partial lesion of both the substantia nigra pars compacta (SNc, A9) and retrorubral field (RRF, A8) on the tremor evoked by harmaline. 6-OHDA (8μg/2μl) was injected unilaterally into the region of the posterior part of the SNc and RRF. Harmaline was administered in a dose of 7.5mg/kg ip on the eighth day after the operation and tremor of forelimbs, head and trunk was measured. We found that the lesion increased intensity of the tremor induced by harmaline but did not influence its character. Stereological examination of the lesion extent revealed losses of dopaminergic (tyrosine hydroxylase-immunoreactive) neurons in the anterior (30%) and posterior (72%) SNc, as well as in RRF (72% on the average). Levels of dopamine and all its metabolites, as well as noradrenaline concentrations, were ipsilaterally moderately decreased in the caudate-putamen in the lesioned animals, however, dopamine and DOPAC in the anterior cerebellum were increased. In the caudate-putamen, the ipsi/contra ratio of dopamine level correlated negatively, while that of dopamine turnover positively with the tremor intensity. However, in the anterior cerebellum an inverse relationship was found. Moreover, this symptom correlated positively with the serotonin level and negatively with the 5-HIAA/serotonin ratio on the contralateral side of the posterior cerebellum. The present results seem to indicate that the modulation of dopaminergic and serotonergic transmissions by the lesion modelling early stages of Parkinsons disease may influence tremor triggered in the cerebellum.

The neostriatal inhibition of catalepsy, but not of muscle rigidity, evoked from the substantia nigra pars reticulata

The effects of a bilateral blockade of neo- and palleostriatal GABAergic mechanisms on catalepsy and muscle rigidity resulting from picrotoxin injection into the substantia nigra pars reticulata (SNR) were studied. The catalepsy and rigidity were induced by a unilateral injection of 100 ng/0.5 microliter of picrotoxin. Bilateral injections of 250 ng/l microliter of picrotoxin into the intermediate-ventral parts of the caudato-putamen (CP) abolished the catalepsy but had no effect on the muscle rigidity induced by an intranigral injection of the drug. Bilateral injections of 250 ng/l microliter of picrotoxin into the globus pallidus (GP) did not influence the catalepsy and rigidity induced by the intranigral injection of the drug. The results indicate that the impulses, connected with the catalepsy evoked from the SNR seem to be transmitted back to the CP and blocked therein by inhibition of GABAergic synapses in its intermediate-ventral part. The impulses, connected with the muscle rigidity evoked from the SNR, presumably do not return to the striatum.