E. Motles

Copper suppresses hippocampus LTP in the rat, but does not alter learning or memory in the morris water maze.

The objective of our study was to determinate the effect of copper on long-term potentiation (LTP) in hippocampus slices and a learning test in the Morris Water Maze (MWM). A group of adult Wistar rats received intraperitoneal (ip) injections of 1 mg/kg of CuSO(4) dissolved in saline for 30 consecutive days (Cu.R). A group of control rats (Sal.R), received saline by the same routes and duration. After this period, every individual of both groups was submitted to learning in MWM. Once the learning was completed, the LTP was studied in slices of hippocampus of both groups. The statistical assessment shows that the rats in both groups did not show significant differences in their progressive learning, notwithstanding that group Cu.R had 14.2 times more copper in their hippocampus and 16.7 times more in the visual cortices than in those of group Sal.R. On the other hand, the neurons of CA1 in hippocampus slices of Sal.R showed a significant development of LTP, but this was not observed in group Cu.R. In a second situation, 13 rats received training in MWM. Then, a group of 6 animals were injected with copper i.p. at the dose and time previously described. The 7 other animals were administered saline. Afterward, both groups were retrained in the MWM. The results obtained in Cu.R were similar to those obtained in Sal.R. Both groups maintained the concentrations of copper in the hippocampus indicated above, nonetheless, only the hippocampus slices of Cu.R did not show LTP. The spatial learning behavior of the rats was not affected by high copper concentration.

Interference of chronically ingested copper in long-term potentiation (LTP) of rat hippocampus

The objective of our study was to find the evidence of copper interaction in LTP, motivated by copper involvement in neurodegenerative illness, like Parkinson, Alzheimer and Amyotrophic Lateral Sclerosis, and we initiated the study of this element in the LTP. For this purpose we used hippocampus slices of rats chronically consuming copper dissolved in water (CuDR; n=26) and non-copper-consuming rats (CR; n=20). The CuDR rats received 8--10 mg/day during 20--25 days. Electrophysiological tests showed absence of LTP in CuDR slices, contrary to CR slices. The stimulus-response test applied before and after LTP showed significant increases of synaptic potential in the CR group. This did not occur in the CuDR group, except for the initial values, which probably seem associated to an early action of copper. The paired-pulse (PP) test, applied to CR and CuDR prior to tetanic stimulation, showed a significant reduction in PP, for the 20-, 30- and 50-ms intervals in CuDR. At the end of the experiments, copper concentration was 54.2 times higher in CuDR slices, compared to the concentration present in CR slices. Our results show that copper reduces synaptic sensibility and also the facilitation capability. These effects represent a significant disturbance in the plasticity phenomenon associated with learning and memory.

Comparative study of the behavioral changes evoked by d-amphetamine and apomorphine in adult cats. Dose-response relationship

The behavioral effects of d-amphetamine and apomorphine administration were studied in 17 adult cats. The doses of amphetamine administered were 0.1, 0.5, 1.0 and 5.0 mg/kg; those of apomorphine, 0.1, 0.5, 1.0 and 2.0 mg/kg. These two drugs evoked in the same animal marked differences in behavioral responses. Amphetamine induced a dose-dependent hypomotility, which was marked with the higher doses. In addition, rhythmic, bilateral slow movements of the head as a mode of stereotypy, indifference to the environment and dose-dependent increase in respiratory rate. Apomorphine elicited limb flicking, dose-dependent hypermotility and increase in olfactory behavior, the last two reactions with stereotypy characteristics. The animals appeared as if being scared, hyperreacting to sudden stimuli and showing total indifference to the surrounding environment. There were marked differences in behavioral responses evoked by these two agonists of the catecholaminergic system. These data do not conform with the behavioral reactions reported in the rat by other investigators. The disagreement with other communications is probably due to differences in reactivity of the species employed. The processes involved in the diversity of the behavioral responses of the cat to the administration of amphetamine and apomorphine have not been delucidated.

Rotational behavior in the cat induced by electrical stimulation of the pulvinar-lateralis posterior nucleus complex: role of the cholinergic system.

We studied the involvement of the cholinergic system in the contralateral head-eye-body turning induced in the cat through stimulation of the pulvinar-lateralis posterior nucleus complex (P-LP). In 17 cats through a cannula aimed at the P-LP, agonists and antagonists of the cholinergic system were injected. The electrical activity of the P-LP could be recorded through the same cannula or through electrodes attached to it. In addition, electrodes were implanted ipsilaterally in the dorsal hippocampus, caudate nucleus, amygdala, and superior colliculus to record through them and through one screw placed on the skull the electrical activity of those structures and of the cortical P-LP projection. Seven days after surgery, carbachol, an agonist of the cholinergic system was injected in the P-LP, and the behavior and electrical activity of the unrestrained cat (previously accustomed to a plastic cage) were recorded. A control volume of 0.9% NaCl was always injected previously. The usual drug volume injected was 1 microliter; occasionally, 2 microliter were injected. Weekly or biweekly sessions were conducted to determine (a) the threshold for cholinergic activation, (b) the threshold for turning behavior, (c) the blocking effect of local atropine sulfate injected previously, (d) the effect of haloperidol previously injected (locally or systemically), and (e) the effect of dioxolane, an exclusive muscarinic agonist. In 14 of 17 cats, contralateral turning behavior was evoked by carbachol. In two of the three cats that did not respond to carbachol, dioxolane induced turning. The effect of dioxolane was similar to that of carbachol when tried in five cats. Besides turning behavior, carbachol produced numerous symptoms due to cholinergic activation. Atropine blocked the rotational effect of carbachol in all cats, and haloperidol blocked it in 68% of them. Electrolytic coagulation of the dorsal hippocampus surrounding the P-LP did not disturb the effects induced by carbachol. These experiments show that both systems of the P-LP, cholinergic and catecholaminergic, are involved in the contralateral turning. We conclude that the effect induced by carbachol is due to activation of muscarinic receptors because it is totally blocked by local atropine sulfate and is reproduced by dioxolane, an exclusive muscarinic agonist.

Effects of p-chloro-phenylalanine on the behaviors induced by apomorphine and amphetamine in adult cats

1. The effects of serotonin depletion on the behaviors evoked by apomorphine or amphetamine are analyzed. Amphetamine (5 mg/kg, s.c.) or apomorphine (2 mg/kg, s.c.) were administered to fourteen adult mongrel cats. Inhibition of tryptophan hydroxylase was achieved by intraperitoneal injection of p-chlorophenylalanine (100 mg/kg daily for three consecutive days). Serotonin depleted animals were tested with either apomorphine or amphetamine (same doses as above). 2. Behaviors evoked by both drugs were recorded and quantified. The following behaviors were rated: motility (locomotion), alertness, fear, indifference, olfaction and lateral head movements. 3. Biochemical analysis of the raphe dorsalis and caudate nuclei of p-CPA treated animals showed an average drop in serotonin concentration of 77%. Serotonin depletion induced statistically significant changes in the following behaviors in amphetamine-treated cats: locomotion, fear, lateral head movements and alertness. Serotonin depleted cats tested with apomorphine showed significant changes only in olfaction and indifference behaviors. 4. Serotonin appears to play a significant modulatory role in some of the behaviors evoked by amphetamine, specially locomotion. Such role is less evident for the behaviors evoked by apomorphine.

Behavioral effects evoked by SKF 38393 and LY 171555 in adult cats

The aim of the present work was to study the behavioral effects elicited in adult cats by the selective D1 agonist, SKF 38393, and the D2 agonist, LY 171555, comparing their effects with those evoked by apomorphine. In 10 adult cats, 0.5, 1.0, 4.0, and 8.0 mg/kg IP of SKF 38393 were administered at random. A dose-response effect was observed related to alertness, indifference, and locomotion. The overall effect of SKF 38393 was inhibitory. To the same 10 animals, LY 171555 in doses of 0.25, 0.5, and 1.0 mg/kg were injected IP. This drug had an excitatory and more complex effect than what was observed with the D1 agonist. Increases in locomotion, in alertness, indifference, fear, olfaction, pupillary dilation, hallucination, limb flicking, and head shaking were recorded. Apomorphine given to the same cats, in a dose equimolar to 1.0 mg/kg of LY 171555, elicited behaviors that resembled those elicited by the latter drug, but of a lesser intensity and duration. The interval between the different treatments was approximately 2 months. These results show clearly that the D2 receptor is the main dopaminergic receptor involved in the mechanism of production of most of the behavioral effects produced by some of the dopaminergic agonist drugs like apomorphine.

Effects of SCH 23390 and sulpiride on the behaviors evoked by amphetamine and apomorphine in adult cats

1. The aim of the present study was to analyze whether the dopaminergic D1 and D2 receptors are involved in the production of the behaviors evoked by parenteral administration of amphetamine and apomorphine in adult cats. 2. Fifteen mongrel cats of both sexes were injected, in separate sessions, with 2.5 mg/kg of amphetamine and 2.0 mg/kg of apomorphine. The D1 receptor blocker, SCH 23390 was administered (0.3 mg/kg i.p.) and after 60 min, amphetamine and apomorphine were again injected on different days. The same procedure was carried on with sulpiride in two doses (20 and 30 mg/kg i.p.). The behaviors induced by the two dopaminergic drugs, before and after the receptor blocker administration were respectively compared. The Wilcoxon signed rank test was employed for statistical analysis. Three independent observers recorded the behaviors. 3. SCH 23390 and sulpiride produced per se hypomotility and sedation, effects that were considered when analysing the results. Some of the behaviors produced by amphetamine (pupillary dilation, head movements) were slightly modified by both receptor blockers. SCH 23390 only modified the licking behavior produced by apomorphine. In contrast, sulpiride blocked almost all the behaviors elicited by apomorphine, especially when the 30 mg/kg dose was administered. It is concluded that the behaviors produced by the 2 mg/kg dose of apomorphine are evoked by its binding to the post-synaptic dopaminergic D2 receptors and blocked by sulpiride.

Cholinergic blockade with scopolamine in adult cats. Effects on the behaviors evoked by apomorphine and amphetamine

1. The aim of this work is to analyse the role that the cholinergic system could play in the production of the behaviors evoked by apomorphine and amphetamine in adult cats. These two drugs were injected s.c. in separate sessions, before and after a s.c. administration of scopolamine which blocked the muscarinic receptors. The pre and post-scopolamine results of the behaviors produced by the two catecholaminergic drugs were compared using the non-parametric Wilcoxon signed rank test. 2. In a previous step a dose-response study of the behavioral effects of scopolamine, in doses of 0.05, 0.1, 0.4 and 0.8 mg/kg was carried out in ten cats. The Kruskal-Wallis and the non-parametric multiple comparison tests were employed. A dose-dependent decrease in motility (locomotion) and a dose-dependent increase in inappetence and pupillary dilation were found. 3. In thirteen cats which were injected with 2 mg/kg of apomorphine and 2.5 mg/kg of amphetamine the findings were: 1--apomorphine after scopolamine produced a decrease in the hypermotility, compared with the results observed with the former drug previous to scopolamine; 2--with amphetamine an increase in immobility and a decrease in indifference were observed. 4. The authors conclude that the decrease in motility recorded with apomorphine and amphetamine after scopolamine, could be attributed to the proper effect of scopolamine. No explanation could be found for the decrease in indifference found by injecting amphetamine after scopolamine. 5. Considering the antagonistic effect between the dopaminergic and the cholinergic systems and that the latter one has an arousal effect, we postulate that the behavioral indifference produced by apomorphine and amphetamine could be the result of a kind of blockade of the cholinergic system when the catecholaminergic system is activated through the administration of the two cited drugs.

Output pathway for turning behavior from the neostriatum and substantia nigra in cats.

The goal of the present work was to study the output pathway of the information for turning behavior originating in the striatum and coursing through the substantia nigra pars reticulata (SNR). In 45 adult cats distributed in 3 groups, Ni-Cr electrodes were implanted in the caudate nucleus and substantia nigra pars reticulata and depending on the animal group in the superior colliculus (SC), nucleus ventralis lateralis/nucleus ventralis medialis (VL/VM) complex or nucleus tegmenti pedunculopontinus (TPP) of one cerebral hemisphere. The threshold current required to evoke turning behavior was determined in each animal for each implantation site. An electrolytic lesion of the superior colliculus, the VL/VM complex or the nucleus tegmenti pedunculopontinus was carried out in each group of cats. The effects of the lesions on behavior and on the electrical threshold currents were determined and compared with the prelesion values. Finally the extent of the lesions and electrode positions were analyzed. The results show that the substantia nigra pars reticulata is the structure with the lowest thresholds for turning and that the superior colliculus appears to be more relevant for carrying the information for turning than either the VL/VM complex or the nucleus tegmenti pedunculopontinus.

Effects of disulfiram, phenoxybenzamine and propranolol on the behaviors evoked by apomorphine and amphetamine in adult cats

The aim of this work was to study the role that the noradrenergic system could play in the mechanism of production of the behaviors evoked by parenteral injection of apomorphine and amphetamine in adult cats. Ten cats were injected s.c. with 2 mg/kg of apomorphine and 2.5 mg/kg of amphetamine in separate sessions. The behaviors were recorded, until control conditions were again attained. In a second stage, disulfiram was administered ip., followed by apomorphine and amphetamine in the same doses as cited above. The effects on behaviors produced by disulfiram and those of apomorphine and amphetamine were recorded by three independent observers. Comparisons of the pre- and post-disulfiram behavioral results were analyzed with the help of the non-parametric Wilcoxon signed rank test. In another group of ten cats a similar procedure was carried on employing the alpha and beta noradrenergic blocking agents, phenoxybenzamine and propranolol. The noradrenergic blocking drugs, especially disulfiram and phenoxybenzamine produced by themselves a decrease in motility, in alertness and an increase in indifference and inappetence. Apomorphine and amphetamine administered after the blocking drugs showed slight behavioural modifications, reflection most of them the changes produced by the three blocking drugs. It is concluded that probably the nor-adrenergic system could be involved in the hypomotility elicited by amphetamine. NA is not involved in the induction of the other behaviors evoked by apomorphine and amphetamine.