Intracerebroventricular Injection Of 6-hydroxydopamine Research Articles

Effects of squalene/squalane on dopamine levels, antioxidant enzyme activity, and fatty acid composition in the striatum of Parkinson^|^rsquo;s disease mouse model

Active oxygen has been implicated in the pathogenesis of Parkinson's disease (PD); therefore, antioxidants have attracted attention as a potential way to prevent this disease. Squalene, a natural triterpene and an intermediate in the biosynthesis of cholesterol, is known to have active oxygen scavenging activities. Squalane, synthesized by complete hydrogenation of squalene, does not have active oxygen scavenging activities. We examined the effects of oral administration of squalene or squalane on a PD mouse model, which was developed by intracerebroventricular injection of 6-hydroxydopamine (6-OHDA). Squalene administration 7 days before and 7 days after one 6-OHDA injection prevented a reduction in striatal dopamine (DA) levels, while the same administration of squalane enhanced the levels. Neither squalene nor squalane administration for 7 days changed the levels of catalase, glutathione peroxidase, or superoxide dismutase activities in the striatum. Squalane increased thiobarbituric acid reactive substances, a marker of lipid peroxidation, in the striatum. Both squalane and squalene increased the ratio of linoleic acid/linolenic acid in the striatum. These results suggest that the administration of squalene or squalane induces similar changes in the composition of fatty acids and has no effect on the activities of active oxygen scavenging enzymes in the striatum. However, squalane increases oxidative damage in the striatum and exacerbates the toxicity of 6-OHDA, while squalene prevents it. The effects of squalene or squalane treatment in this model suggest their possible uses and risks in the treatment of PD.

Selective inhibition of monoamine oxidase A or B reduces striatal oxidative stress in rats with partial depletion of the nigro-striatal dopaminergic pathway

Partial lesion (50%) of the nigro-striatal dopaminergic pathway induces compensatory increase in dopamine release from the remaining neurons and increased extracellular oxidative stress (OS-ec) in the striatum. The present study was designed to explore the role of monoamine oxidase types A and B (MAO-A, MAO-B) in producing this increased oxidative stress. Lesion of the dopaminergic pathways in the CNS was produced in rats by intra-cerebroventricular injection of 6-hydroxydopamine (6-OHDA; 250 μg) and striatal microdialysis was carried out 5 weeks later. Striatal OSec was determined by measurement of oxidized derivatives of the marker molecule N-linoleyl-tyrosine. Striatal tissue MAO-A activity was unchanged by 6-OHDA lesion but MAO-B activity was increased by 16%, together with a 45% increase in glial cell content. The selective MAO-B inhibitor rasagiline (0.05 mg/kg s.c. daily for 14 days) did not affect microdialysate dopamine concentration [DAec] in sham-operated rats, but decreased OSec by 30%. In lesioned rats, rasagiline decreased [DAec] by 42% with a 49% reduction in OSec. The decrease in [DAec] was reversed by the dopamine D2 receptor antagonist sulpiride (10 mg/kg s.c.). The selective MAO-A inhibitor clorgyline (0.2 mg/kg s.c. daily for 14 days) increased striatal [DAec] by 72% in sham-operated rats with no change in OSec. In lesioned rats clorgyline increased [DAec] by 66% and decreased OSec by 44%. Rasagiline and clorgyline were effective to a similar extent in reduction of tissue levels of 7-ketocholesterol and the ratio GSSG/GSH, indicative of reduced intracellular oxidative stress level. This data implies that gliosis in our 6-OHDA animals together with inhibition of glial cell MAO-B by rasagiline causes an increase in local levels of dopamine at the presynaptic receptors, and a reduction in dopamine release (and in [DAec]) by presynaptic inhibition. Moreover, inhibition of MAO-A or MAO-B reduces the enhanced level of oxidative stress in the lesioned striatum, and while both clorgyline and rasagiline reduced DA oxidative metabolism, rasagiline possesses an additional antioxidant property, not only that resulting from MAO inhibition.

Enhanced striatal glutamate release after the administration of rimonabant to 6-hydroxydopamine-lesioned rats

While recent studies have shown that the blockade of cannabinoid CB(1) receptors might be beneficial to alleviate the motor inhibition typical of Parkinson's disease (PD), the neurochemical substrates for this effect remain elusive. Here we have carried out microdialysis experiments to determine whether the effects of rimonabant, a selective antagonist of CB(1) receptors, might be associated with changes in striatal glutamate release in a rat model of PD generated by intracerebroventricular injection of 6-hydroxydopamine. Our data demonstrate that the treatment with rimonabant slightly increased striatal glutamate release in control rats, although this effect was only evident with the highest dose of rimonabant tested (1mg/kg). However, the increase in glutamate release was much more marked in the parkinsonian rats where similar changes were observed at a dose of 1 and 0.1mg/kg, exactly the same dose that relieved motor inhibition in previous behavioral studies. In summary, the potential of rimonabant to act as a possible antihypokinetic agent in parkinsonian rats seems to be related to enhanced glutamate release from excitatory afferents to the striatum. This observation is of potential clinical interest, particularly for those parkinsonian patients that exhibit a poor response to classic levodopa treatment.

Effects of rimonabant, a selective cannabinoid CB 1 receptor antagonist, in a rat model of Parkinson's disease

Recent evidence suggest that the blockade of cannabinoid CB 1 receptors might be beneficial to alleviate motor inhibition typical of Parkinson's disease (PD). In the present study, we have explored the motor effects of rimonabant, a selective antagonist of CB 1 receptors, in a rat model of PD generated by an intracerebroventricular injection of 6-hydroxydopamine. Compared with rats subjected to unilateral injection of this toxin in the medial forebrain bundle, this model allows nigral dopaminergic neurons be symmetrically affected. Dose-response studies with 6-hydroxydopamine revealed that the application of 200 μg per animal caused hypokinetic signs (decreased ambulatory activity, increased inactivity, and reduced motor coordination), which paralleled several signs of degeneration of nigrostriatal dopaminergic neurons (dopamine depletion in the caudate-putamen, and decreased mRNA levels for tyrosine hydroxylase and superoxide dismutase-1 and -2 in the substantia nigra). In these conditions, the degree of hypokinesia and dopaminergic degeneration may be considered moderate, comparable to the disturbances occurring in early and middle stages of PD in humans, a period that might be appropriate to test the effects of rimonabant. There is also degeneration of other dopaminergic pathways out of the basal ganglia, but this does not appear to interfere significantly with the hypokinetic profile of these rats. Higher doses of 6-hydroxydopamine elevated significantly animal mortality and lower doses failed in general to reproduce motor inhibition. Like other animal models of PD, these rats exhibited an increase in the density of CB 1 receptors in the substantia nigra, which is indicative of the expected overactivity of the cannabinoid transmission in this disease and supports the potential of CB 1 receptor blockade to attenuate hypokinesia associated with nigral cell death. Thus, the injection of 0.1 mg/kg of rimonabant partially attenuated the hypokinesia shown by these animals with no effects in control rats, whereas higher doses (0.5–1.0 mg/kg) were not effective. We also found that the antihypokinetic effects of low doses of rimonabant did not influence the dopamine deficits of these animals, as well as it did not modify GABA or glutamate transmission in the caudate-putamen. In summary, rimonabant may have potential antihypokinetic activity in moderate parkinsonism at low doses, but this effect is not related to changes in dopaminergic, GABAergic, or glutamatergic transmission in the striatum. Therefore, the elucidation of the neurochemical substrate involved in this effect remains a major challenge for the future.

Central and peripheral catecholamines regulate the exercise-induced elevation of plasma interleukin 6 in rats

Several recent reports indicate that exercise elevates the plasma interleukin 6 levels; however, the precise regulation of such an elevation still remains to be clarified. In this study, in order to clarify the requirements of central and peripheral catecholaminergic system for this exercise-induced interleukin 6 elevation, rats were either intraperitoneally or intracerebroventricularly injected with 6-hydroxydopamine which depletes the catecholamine in the central or peripheral tissues. As a result, our exercise protocol elevated the plasma interleukin 6, ACTH, and corticosterone levels in response to exercise. All such exercise-induced increases in the interleukin 6, ACTH, and corticosterone levels were significantly inhibited by pretreatment with an intracerebroventricular injection of 6-hydroxydopamine. In the intraperitoneal 6-hydroxydopamine-treated animals, the exercise-induced interleukin 6 elevation was significantly suppressed compared with the vehicle-treated animals, although no significant difference was found in either the ACTH level or the corticosterone level between both groups of animals. These results thus suggest that central and peripheral catecholamines are involved in the regulation of the exercise-induced interleukin 6 elevation.

Motor behavioural changes after intracerebroventricular injection of 6-hydroxydopamine in the rat: an animal model of Parkinson's disease

At the beginning of the 1970s, different studies reported behavioural disturbances after the intracerebroventricular (icv) administration of 6-hydroxydopamine (6-OHDA) in the rat. Despite the fact that this neurotoxic agent degenerates brain dopaminergic (DA-) cells, its potential utility to produce a rat model of Parkinson's disease (PD) was never systematically studied because the aphagia and adipsia were often observed. In the present study, a procedure that induces a marked DA-cell degeneration that bypasses these and other undesirable complications of icv injection of 6-OHDA is reported. Lesioned animals (50–500 μg of 6-OHDA) showed a persistent motor syndrome composed of hypokinesia, purposeless chewing and catalepsy. The intensity of motor signs was dose-dependent, and recovered partially after administration of DA-receptor agonists, exposure to sensorial stimuli and stress, three procedures that reduce motor dysfunctions in Parkinson's disease (PD). Lesioned animals showed bilateral and symmetrical midbrain DA-cell degeneration with the highest cell-loss in A9 group (substantia nigra), followed by A8 (retrorubral field) and A10 (ventral tegmental area) groups. The similarity between the behavioural syndrome and the topographical profile of cell-loss after icv injection of 6-OHDA in rats and the clinical and neuropathological features of PD indicates that this may be a convenient animal model of PD particularly useful for checking in rats the possible efficacy of new anti-parkinsonian drugs on specific parameters of motor dysfunctions.

Attenuation of 6-hydroxydopamine-induced dopaminergic nigrostriatal lesions in superoxide dismutase transgenic mice

6-Hydroxydopamine is a neurotoxin that produces degeneration of the nigrostriatal dopaminergic pathway in rodents. Its toxicity is thought to involve the generation of superoxide anion secondary to its autoxidation. To examine the effects of the overexpression of Cu,Zn-superoxide dismutase activity on 6-hydroxydopamine-induced dopaminergic neuronal damage, we have measured the effects of 6-hydroxydopamine on striatal and nigral dopamine transporters and nigral tyrosine hydroxylase-immunoreactive neurons in Cu,Zn-superoxide dismutase transgenic mice. Intracerebroventricular injection of 6-hydroxydopamine (50 μg) in non-transgenic mice produced reductions in the size of striatal area and an enlargement of the cerebral ventricle on both sides of the brains of mice killed two weeks after the injection. In addition, 6-hydroxydopamine caused marked decreases in striatal and nigral [ 125I]RTI-121-labelled dopamine transporters not only on the injected side but also on the non-injected side of non-transgenic mice; this was associated with decreased cell number and size of tyrosine hydroxylase-immunoreactive dopamine neurons in the substantia nigra pars compacta on both sides in these mice. In contrast, superoxide dismutase transgenic mice were protected against these neurotoxic effects of 6-hydroxydopamine, with the homozygous transgenic mice showing almost complete protection. These results provide further support for a role of superoxide anion in the toxic effects of 6-hydroxydopamine. They also provide further evidence that reactive oxygen species may be the main determining factors in the neurodegenerative effects of catecholamines.

Interactions between D1 and muscarinic receptors in the induction of striatal c- fos in rats depleted of dopamine as neonates

The contributions of striatal D1 receptors to the expression of sensorimotor behavior are qualitatively different in rats depleted of dopamine (DA) as neonates vs. as adults. In an effort to reveal neuronal mechanisms underlying these behavioral differences we determined the effects of the partial D1 agonist SKF 38393, the muscarinic antagonist scopolamine, and the combination of the two drugs on the induction of c- fos in the striatum and its projection sites, the globus pallidus and substantia nigra. Adult rats, given intracerebroventricular injections of 6-hydroxydopamine (6-OHDA, 50 μg/5 μl/hemisphere) or its vehicle on postnatal day 3, were treated with SKF 38393 (1.5 mg/kg, i.p.), scopolamine (5.0 mg/kg, i.p.) or the combination of the two drugs. There was no significant induction of c- fos in vehicle-treated controls, regardless of drug administration. In DA-depleted rats, scopolamine also did not induce c- fos whereas SKF 38393 produced a significant increase in the number of FOS-positive cells in the dorsal, but not ventral, striatum. The combined administration of scopolamine and SKF 38393 resulted in a potent synergism in the number of FOS-positive cells in DA-depleted rats. These interactions between lesion condition and drugs on c- fos induction were not secondary to differences in drug-induced behavioral activity. Activity levels were no different in vehicle vs. DA-depleted rats following the combined administration of scopolamine + SKF 38393, yet the two groups of rats exhibited marked differences in the density of FOS-positive striatal neurons. The effects of scopolamine and SKF 38393 on c- fos induction in striatum are qualitatively similar to those reported in rats DA-depleted as adults and suggest that, at this single-label level of analysis, the ability of D1 and muscarinic receptors to influence striatal activity does not contribute to the marked age-related differences in the behavioral effects of DA depletions.

S-51-2 - Potentials of MAO-A and B inhibitors in the clinic

Partial lesion (50%) of the nigro-striatal dopaminergic pathway induces compensatory increase in dopamine release from the remaining neurons and increased extracellular oxidative stress (OS-ec) in the striatum. The present study was designed to explore the role of monoamine oxidase types A and B (MAO-A, MAO-B) in producing this increased oxidative stress. Lesion of the dopaminergic pathways in the CNS was produced in rats by intra-cerebroventricular injection of 6-hydroxydopamine (6-OHDA; 250 μg) and striatal microdialysis was carried out 5 weeks later. Striatal OSec was determined by measurement of oxidized derivatives of the marker molecule N-linoleyl-tyrosine. Striatal tissue MAO-A activity was unchanged by 6-OHDA lesion but MAO-B activity was increased by 16%, together with a 45% increase in glial cell content. The selective MAO-B inhibitor rasagiline (0.05 mg/kg s.c. daily for 14 days) did not affect microdialysate dopamine concentration [DAec] in sham-operated rats, but decreased OSec by 30%. In lesioned rats, rasagiline decreased [DAec] by 42% with a 49% reduction in OSec. The decrease in [DAec] was reversed by the dopamine D2 receptor antagonist sulpiride (10 mg/kg s.c.). The selective MAO-A inhibitor clorgyline (0.2 mg/kg s.c. daily for 14 days) increased striatal [DAec] by 72% in sham-operated rats with no change in OSec. In lesioned rats clorgyline increased [DAec] by 66% and decreased OSec by 44%. Rasagiline and clorgyline were effective to a similar extent in reduction of tissue levels of 7-ketocholesterol and the ratio GSSG/GSH, indicative of reduced intracellular oxidative stress level. This data implies that gliosis in our 6-OHDA animals together with inhibition of glial cell MAO-B by rasagiline causes an increase in local levels of dopamine at the presynaptic receptors, and a reduction in dopamine release (and in [DAec]) by presynaptic inhibition. Moreover, inhibition of MAO-A or MAO-B reduces the enhanced level of oxidative stress in the lesioned striatum, and while both clorgyline and rasagiline reduced DA oxidative metabolism, rasagiline possesses an additional antioxidant property, not only that resulting from MAO inhibition.

Rapid and long lasting reduction of the hypothermic effect of a D2 dopamine agonist after an intracerebroventricular injection of 6-hydroxydopamine

In mice pretreated intracerebroventricularly (i.c.v.) with 6-hydroxydopamine (6OHDA) (50 μg per mouse), as soon as the hypothermia elicited by the neurotoxin had vanished (3 hr), the hypothermic effect induced by the direct D2 dopamine receptor agonist RU 24926 (1 mg/kg, s.c.), was almost completely suppressed. This reduction in hypothermic effect was observed more than 1 month after the 6OHDA injection. On the 3rd day after 6OHDA injection, this reduction was observed for all tested doses of RU 24926 (0.25–2 mg/kg). It was prevented when an i.p. administration of the norepinephrine uptake inhibitor desipramine (20 mg/kg) was performed 30 min before the 60HDA i.c.v. injection. It was not modified when an i.p. administration of the dopamine uptake inhibitor GBR 12783 (20 mg/kg) was performed 30 min before the 6OHDA i.c.v. injection. The 6OHDA i.c.v. injection modified significantly neither the dopamine nor the serotonin hypothalamic contents. On the contrary it resulted in a marked decrease (− 73%) of the norepinephrine hypothalamic content, which was unchanged by the administration of GBR 12783 (20 mg/kg, i.p.) 30 min before 6OHDA, but completely prevented by desipramine (20 mg/kg, i.p.) 30 min before 6OHDA i.c.v. injection. It is concluded that the hypothermic effect resulting from the stimulation of D2 dopamine receptors involves a central norepinephrine transmission, which is very rapidly altered after 6OHDA administration.

Changes in lipid peroxidation, Cu/Zn-superoxide dismutase and its mRNA following an intracerebroventricular injection of 6-hydroxydopamine in mice

A single i.c.v. injection of 6-hydroxydopamine (6-OHDA) in mice resulted in a biphasic increase in lipid peroxidation as assayed by the level of thiobarbituric acid-reacting substances (TBARS). An increase in Cu/Zn-superoxide dismutase (SOD) activity was temporally related with the first peak of TBARS but remained unchanged during the second TBARS peak. This suggests that a free radical species other than O 2 − may be involved in the late onset increase in TBARS. The level of Cu/Zn-SOD mRNA did not immediately reflect the change in Cu/Zn-SOD activity but rather increased gradually reaching significantly higher levels only 8 weeks after i.c.v. an injection of 6-OHDA. This increase in Cu/Zn-SOD mRNA likely occurs in response to a consumption of intrinsic SOD. Thus, short- and long-term increases in lipid peroxidation likely occur by different mechanisms and studies of both are needed to elucidate the neurodegenerative process.

Light-microscopic localization of somatostatin binding sites in the locus coeruleus of the rat

Somatostatin (SS14) binding sites within locus coeruleus (LC) were localized at the light microscope level by [ 125I][Tyr 0, d-Trp 8]SS14 radioautography combined with an immunohistochemical/neurotoxic lesioning approach. In intact rats, the dense accumulation of SS14 binding sites of LC conspicuously overlapped with the cluster of tyrosine hydroxylase (TH) immunoreactive neurons; SS14 specific binding was directly proportional to the number of TH immunostained (TH+) cell bodies per mg of tissue throughout LC. Complete lesion of catecholaminergic nerve cell bodies of LC by intracerebroventricular injection of 6-hydroxydopamine (6-OHDA) resulted in the total abolition of SS14 specific binding in the structure. In addition, specifically bound [ 125I][Tyr 0, d-Trp 8]SS14 and TH+ cell density were quantified serially in a set of rats bearing various partial neurotoxic lesions; a highly significant correlation was found between the two parameters at each of the 16 coronal levels of LC examined. The coefficient of proportionality was identical at all levels. These results strongly suggest that somatostatin binding sites are uniformly localized on all noradrenergic neurons of LC.

Noradrenaline-induced emesis: Alpha-2 adrenoceptor mediation in the area postrema

The emetic action of noradrenaline was investigated in unanaesthetized cats, after it was injected into the cerebral ventricles through chronically implanted cannulae. Intracerebroventricular injection of noradrenaline produced dose-dependent and shortlasting emesis, which was abolished after ablation of the area postrema. However, copper sulphate, given orally, evoked emesis in cats with an ablated area postrema. The selective alpha-2 adrenoceptor antagonist, yohimbine, as well as the mixed alpha-1 and alpha-2 adrenoceptor blocking drugs, phentolamine, tolazoline, phenoxybenzamine and dihydroergotamine, but not the selective alpha-1 adrenoceptor antagonist, prazosin, all injected into the cerebral ventricles, attenuated or blocked the emesis evoked by intracerebroventricular injection of noradrenaline. Of the alpha-adrenoceptor antagonist, only yohimbine produced dose-dependent inhibition of the emesis induced by noradrenaline. On the contrary, selected beta-adrenoceptor blocking agents, an antimuscarinic drug, a ganglionic blocking agent, an antihistamine, dopamine antagonists and a 5-hydroxytryptamine antagonist, all injected into the cerebral ventricles, had no significant effect on the emesis induced by noradrenaline, similarly injected. The emetic response to intracerebroventricular injection of noradrenaline, as well as to intragastric adminsitration of copper sulphate was not altered in cats pretreated with intracerebroventricular injections of alpha-methyl- p-tyrosine and bretylium. On the other hand, the emetic response to intracerebrocentricular injection of noradrenaline and to intragastric administration of copper sulphate was attentuated or blocked in cats pretreated with reserpine intracerebroventricularly. Moreover, in cats pretreated with intracerebroventricular injection of 6-hydroxydopamine and hemicholinium, the emesis induced by intracerebroventricular administration of noradrenaline but not that produced by intragastric injection of copper sulphate, was depressed. Finally, in cats pretreated with an intracerebroventricular injection of 5,6-dihydroxytryptamine, the emetic response to intracerebroventricular injection of noradrenaline as well as to intragastric administration of copper sulphate was not changed. It is postulated that the emesis produced by noradrenaline, injected into the cerebral ventricles, is mediated through alpha-2 adrenoceptors. These alpha-2 adrenoceptors appear to be located postsynaptically in the area postrema and they transmit emetic impulses from the area postrema to the emetic region of the brainstem reticular formation. The results obtained with reserpine and copper sulphate indicate that monoamines are involved in the regulation of vomiting in the emetic region of the brainstem reticular formation.

O-197 - Immune responses in mice treated with Intracerebroventricular injection of 6-hydroxydopamine or 5,7-dihydroxytryptamine

O-197 - Immune responses in mice treated with Intracerebroventricular injection of 6-hydroxydopamine or 5,7-dihydroxytryptamine

Cardiovascular and sympathetic responses to intracarotid and intravenous injections of serotonin in rats.

The possible central regulation of cardiovascular system by serotoninergic neurones was investigated in rats by using a cross-circulated head preparation. Intracarotid injections of serotonin (5-HT, 0.1-10 micrograms) consistently produced neurally mediated vasodepression in the recipient body. Although intravenous injections of 5-HT (0.1-1 micrograms) caused similar depressor responses, larger doses (5-10 micrograms) caused biphasic responses, consisting of vasodepression followed by a sustained pressor response. Despite the biphasic blood pressure responses, sympathetic nerve activity was consistently reduced by the intravenous 5-HT. Bilateral vagotomy abolished the vasodepressor responses to intravenous injections of 5-HT (10 micrograms) and made the responses purely pressor, but the vasodepressor responses to intracarotid injections of 5-HT (10 micrograms) were largely unaffected. Following destruction of central noradrenergic neurones by intracerebroventricular injections of 6-hydroxydopamine, the vasodepressor responses to intracarotid 5-HT (1-10 micrograms), but not to the intravenous 5-HT, were significantly reduced. These results suggest that intravenous injections of 5-HT reduce nerve activity by affecting the reflex mechanism via the vagus nerves. On the other hand, the vasodepression and reduction of the nerve activity by the intracarotid injections appears to be of central origin, and the vasodepression could be mediated via noradrenergic mechanisms in the brain.

Lesioning of serotoninergic and noradrenergic nerve fibres of the rat brain does not decrease binding of 3H-clonidine and 3H-rauwolscine to cortical membranes.

alpha 2-Adrenoceptors located presynaptically on nerve terminals are known to modulate the release of neurotransmitters from noradrenergic and serotoninergic neurons. The pre- and/or postsynaptic localization of binding sites for alpha 2-adrenergic radioligands, the agonist 3H-clonidine and the antagonist 3H-rauwolscine, was investigated in the rat cerebral cortex by the use of specific neurotoxins. Intracerebroventricular injections of 6-hydroxydopamine (6-OH-DA) and 5,7-dihydroxytryptamine (5,7-DHT) were used to destroy the noradrenergic and serotoninergic neurons, respectively, and the success of the treatment was controlled by measurement of tritium accumulation in cortex slices incubated with 3H-noradrenaline or 3H-serotonin. In cortical membranes, 3H-rauwolscine bound to a single site (KD about 5 nmol/l; Bmax 217-247 fmoles/mg protein), whereas 3H-clonidine bound to a high affinity site (KD 0.6-1.4 nmol/l) and a low affinity site (KD 6-10 nmol/l). The total number of high plus low affinity 3H-clonidine binding sites was about two thirds of the number of 3H-rauwolscine binding sites. 6-OH-DA treatment significantly increased the number of high affinity 3H-clonidine binding sites without reducing the number of high plus low affinity binding sites, indicating a denervation supersensitivity. KD- as well as Bmax-values for 3H-rauwolscine remained unaltered after 6-OH-DA-treatment. Since an increase in postsynaptic alpha 2-adrenoceptors due to 6-OH-DA-administration might have masked a loss of presynaptic alpha 2-adrenergic binding sites, rats were chronically treated with high doses of clonidine in order to prevent a possible supersensitivity of postsynaptic receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Augmented central cholinergic mechanisms in spontaneously hypertensive rats. Involvement of deranged noradrenergic mechanisms in the brain.

Intracerebroventricular (ICV) injections of carbachol produced biphasic blood pressure responses consisting of initial vasodepression of short duration followed by a sustained pressor phase, which were accompanied by corresponding changes in sympathetic nerve activity in normotensive outbred-Wistar rats (NT) under urethane-anesthesia. In both normotensive Kyoto Wistar rats (WKY) and spontaneously hypertensive rats (SHR), on the other hand, carbachol elicited purely pressor responses, and accompanying sympathetic nerve activity was little affected. The magnitude of the pressor responses was larger in SHR than in WKY or NT rats. Spinal sectioning did not affect the magnitude of the pressor responses. Vasopressor responses to intravenous injections of arginine-vasopressin were not significantly different between WKY and SHR. These results indicate that carbachol injected intracerebroventricularly produces vasopressor effects mainly by releasing pituitary hormones, probably vasopressin, and that augmented pressor responses in SHR may be due to excessive release of vasopressin. When central noradrenergic neurons had been destroyed with ICV injections of 6-hydroxydopamine in both NT and WKY rats, carbachol-induced vasopressor responses were markedly augmented and resulted in responses similar to those of SHR. These findings indicate that central noradrenergic vasodepressive neurons are deficient and that the augmented vasopressor responses to carbachol resulted from deranged central noradrenergic mechanisms in SHR.

Central dopaminergic regulation of adrenomedullary ornithine decarboxylase activity

The administration of the two dopamine receptor agonists apomorphine (APM) and piribedil (PBD) to rats leads to an increase in ornithine decarboxylase (ODC) activity in the adrenal medulla. In this work, we have tried to elucidate the neural pathways involved in the regulation of this enzyme. The treatments used are: unilateral splanchnicotomy, spinal cord section, intraventricular injection of the neurotoxin 6-hydroxydopamine and section of the brain at various levels. Unilateral splanchnicotomy reduces very significantly the induction of ODC produced by either APM or PBD. Spinal cord section at either of two different levels (T 5 or T 2) also lowers the response to APM. Intracerebroventricular injection of 6-hydroxydopamine, on the other hand, elevates the mean response to APM, although not to a statistically significant extent. Section of the mesencephalon well below the periaqueductal gray does not alter the response of adrenomedullary ODC to APM. Transection of the diencephalon almost prevents it whereas hypothalamic deafferentation and incomplete diencephalic transection potentiate the effect of this drug. These observations strongly suggest that adrenomedullary ODC activity is predominantly regulated by a central system, originating mainly in the diencephalon-telencephalon and including a facilitatory dopaminergic component.

Effects of dopamine-depleting brain lesions on experimental hyperphagia in rats

Dopamine (DA)-depleting brain lesions of various sizes were produced in rats either by intracerebroventricular injections of 6-hydroxydopamine (6-HDA) or by electrolytic lesions of the lateral hypothalamic (LH) area. Among 30 animals that became aphagic and adipsic for at least four days after large LH or 6-HDA-induced brain lesions, only three developed hyperphagia after electrolytic lesions of the ventromedial hypothalamus (VMH) or daily injections of long-acting protamine-zinc insulin (PZI). In 20 rats with smaller LH or 6-HDA-induced lesions, which had not shown marked initial behavioral dysfunctions, only three gained as much weight after VMH lesions as the control animals. Similarly, 6 of 10 rats with smaller LH lesions could not tolerate a 15-day series of PZI treatments, although 14 of 17 rats with smaller 6-HDA-induced lesions increased their food intake and gained weight during the PZI treatments as did control animals. These results indicate that hypothalamic hyperphagia can be blocked by DA-depleting brain lesions that neither produce an initial period of aphagia and adipsia nor involve hypothalamic tissue. They further indicate that even small LH lesions may prevent the development of hyperphagia elicited by PZI, whereas only very large 6-HDA-induced lesions consistently have this effect.

EFFECT OF INTRACEREBROVENTRICULAR ADMINISTRATION OF CHLORPROMAZINE ON THE SERUM LEVEL OF FREE FATTY ACIDS IN RATS

AbstractThe effect of intracerebroventricular administration of chlorpromazine (CPZ) on the serum level of free fatty acid (FFA) was studied in rats. Injection of CPZ into the lateral ventricle caused a transient decrease in serum FFA, showing the lowest level after 30 min. This decrease in serum FFA caused by CPZ was significantly inhibited by simultaneous injection of dopamine or apomorphine, although noradrenaline and serotonin had no effect. The dopaminergic blocking agent haloperidol caused a rapid decrease in serum FFA. After central chemical sympathectomy with the intracerebroventricular injection of 6-hydroxydopamine. the response to CPZ of serum FFA was completely abolished; but after peripheral sympathectomy by i.v. injection of 6-hydroxydopamine, a partial inhibition of the action of CPZ was shown. These results suggest a possible involvement of the central dopaminergic mechanism in the decrease in serum FFA after intracerebroventricular injection of CPZ.