Carlos Tomaz

The elevated T-maze: A new animal model of anxiety and memory

In an attempt to analyze different types of anxiety, and at the same time assess memory, a new experimental model was developed. The apparatus, named the elevated T-maze, consisted of three arms of equal dimensions (50 x 10 cm) elevated 50 cm from the ground. One arm, enclosed by 40-cm high walls, was perpendicular to two open arms. The first experimental session was conducted 25 min after IP injection of either drug or saline. To assess inhibitory (passive) avoidance, the rat was placed at the end of the enclosed arm and the time taken to withdraw from this arm was recorded three times in succession. Soon afterwards, the rat was placed at the end of one of the open arms and the time taken to withdraw from this arm was measured, thus estimating one-way escape. To assess memory, inhibitory avoidance and escape were measured again 3 days later, without drug. Dose-response curves were determined for the benzodiazepine anxiolytic and amnestic agent diazepam (DZP, 0.5-4 mg/kg), as well as for ipsapirone (IPS, 0.25-2 mg/kg), an azapirone anxiolytic that is devoid of clinically significant amnestic effects. The doses of 1, 2, and 4 mg/kg DZP and of 1 and 2 mg/kg IPS impaired inhibitory avoidance, an effect that may be viewed as anxiolytic. Inhibitory avoidance remained impaired 3 days later in the rats treated with 1-4 mg/kg DZP, indicating anterograde amnesia. This effect was not due to state-dependent learning, because rats injected both at pretraining and pretesting with 2 mg/kg DZP still showed complete amnesia.(ABSTRACT TRUNCATED AT 250 WORDS)

Anxiolytic-like action of neurokinin substance P administered systemically or into the nucleus basalis magnocellularis region

There is evidence that the neurokinin substance P plays a role in neural mechanisms governing learning and reinforcement. Reinforcing and memory-promoting effects of substance P were found after it was injected into several parts of the brain and intraperitoneally. With regard to the close link between anxiety and memory processes for negative reinforcement learning, the aim of the present study was to gauge the effect of substance P on anxiety-related behaviors in the rat elevated plus-maze and social interaction test. Substance P was tested at injection sites where the neurokinin has been shown to promote learning and to serve as a reinforcer, namely in the periphery (after i.p. administration) and after injection into the nucleus basalis magnocellularis region. When administered i.p., substance P had a biphasic dose-response effect on behavior in the plus-maze with an anxiolytic-like action at 50 microg/kg and an anxiogenic-like one at 500 microg/kg. After unilateral microinjection into the nucleus basalis magnocellularis region, substance P (1 ng) was found to exert anxiolytic-like effects, because substance P-treated rats spent more time on the open arms of the plus-maze and showed an increase in time spent in social interaction. Furthermore, the anxiolytic effects of intrabasalis substance P were sequence-specific since injection of a compound with the inverse amino acid sequence of substance P (0.1 to 100 ng) did not influence anxiety parameters. These results show that substance P has anxiolytic-like properties in addition to its known promnestic and reinforcing effects, supporting the hypothesis of a close relationship between anxiety, memory and reinforcement processes.

Evidence for the involvement of serotonin in the antinociception induced by electrical or chemical stimulation of the mesencephalic tectum.

A great deal of evidence has shown that electrical stimulation or microinjections of GABAA blockers, such as bicuculline, into the midbrain tectum (MT) produce escape behavior, which has been associated to fear. This study was aimed to examine the characteristics of the analgesia that follows the escape behavior induced by electrical (freezing and escape thresholds) and chemical (bicuculline microinjections) stimulation of the midbrain tectum. Immediately after the expression of the aversive responses the rats were submitted to the tail-flick test. The obtained results show that analgesia always follows aversive responses integrated at the MT level regardless of the kind of stimulation applied. The antinociceptive effects induced by either electrical or chemical stimulation of the MT were not antagonized by central microinjections of naloxone. On the other hand, the non-specific serotonin antagonist methysergide microinjected into the MT was effective in antagonizing the analgesia induced by any of the aversive stimulations. Based on these results we suggest that serotonin, but not opioid mechanisms, may be involved in the integration of antinociceptive responses to stimulation of the midbrain tectum.

Increased training in an aversively motivated task attenuates the memory-impairing effects of posttraining N-methyl-D-aspartate-induced amygdala lesions

This study was designed to examine the effect of variations in the amount of preoperative training on the retention deficit produced by posttraining lesions of the amygdaloid complex (AC). Rats received 1, 10, or 20 training trials in a footshock-motivated escape task 7 days before receiving N-methyl-D-aspartate lesions of the AC. Inhibitory avoidance retention performance, which was measured 4 days postoperatively, indicated that increased training improved retention in AC-lesioned animals as well as in control animals. The retention performance of AC-lesioned animals was impaired when compared with that of controls; however, the impairment was partially attenuated by increased preoperative training. The finding that AC-lesioned animals displayed greater locomotor activity on the retention test compared with nonlesioned controls suggests that the increased activity may have contributed to the impaired inhibitory avoidance retention performance. Two days after the retention test, some of the AC-lesioned animals were subsequently trained on a continuous multiple-trial inhibitory avoidance response in the same apparatus. AC lesions did not block acquisition or retention of the task. These findings suggest that the amygdala may not be a critical site for the permanent changes mediating stimulus-affect associations based on extensive training.

Localization in the amygdala of the amnestic action of diazepam on emotional memory

It is well known that systemically administered benzodiazepines (BZDs) induce anterograde amnesia in a variety of learning tasks. BZs effects are mediated through the GABAA complex by enhancing GABA-induced synaptic inhibition. As the GABAergic system in the amygdaloid complex (AC) is a site of action for the anxiolytic effects of BZs, such findings suggest that BZs may also influence memory through the amygdala. The present report summarizes a recent series of experiments designed to examine this implication. In a first experiment rats received either sham or bilateral AC lesion using N-methyl-D-aspartic acid (NMDA). One week later, animals were trained on an inhibitory avoidance task and tested 48 h later. Diazepam (DZP; 1.0 and 2.0 mg/kg, i.p.) or vehicle was injected 30 min prior to acquisition. The results demonstrate that DZP-induced retention deficits was blocked in rats with AC lesions. In a second experiment, in an attempt to localize the site of BZDs amnestic action in the AC, we tested the effects of DZP in rats with bilateral ibotenic acid-induced lesions of central (CE), lateral (LAT) or basolateral (BL) amygdala nuclei. The results shown that retention was impaired in animals with CE and LAT lesions but not in animals with BL lesions. In a third experiment we tested the effects of DZP microinjections in different nuclei of the AC on retention performance of rats trained in an avoidance task. The results demonstrate that DZP microinjection prior training in the BL/LAT, but not CE nuclei produce anterograde amnesia.(ABSTRACT TRUNCATED AT 250 WORDS)

Increased neostriatal dopamine activity after intraperitoneal or intranasal administration of L-DOPA: on the role of benserazide pretreatment.

L‐DOPA provides the most potent medication to treat Parkinsons disease, and such systemic treatment is usually combined with a peripheral amino acid decarboxylase inhibitor to amplify its central effectiveness. Since L‐DOPA can lose its efficacy or can lead to adverse effects with prolonged application, current pharmacokinetic and dynamic research is aimed at improving the drugs applicability. In a previous study, performed with in vivo microdialysis in the anesthetized rat, we have shown that intranasal L‐DOPA administration (without prior decarboxylase inhibition) can increase extracellular dopamine levels in the neostriatum. Using similar experimental conditions in the present experiment, we tested the neurochemical effects of L‐DOPA treatment in combination with the peripheral amino acid decarboxylase inhibitor benserazide. In accordance with other data, it was found that the combination of i.p. benserazide and i.p. L‐DOPA led to pronounced increases of extracellular levels of dopamine, dihydroxyplenylacetic acid and homovanillic acid in the neostriatum, whereas i.p. L‐DOPA alone only moderately increased dopamine, but strongly increased the metabolite levels. Furthermore, increased dopamine levels, and weaker increases of dihydroxyplenylacetic acid and homovanillic acid were observed after i.p. benserazide followed by intranasal L‐DOPA. Finally, we found that i.p. benserazide alone can lead to pronounced increases in neostriatal dopamine and moderate increases of dihydroxyplenylacetic acid levels, whereas it did not affect homovanillic acid. Thus, not only the combination of L‐DOPA (i.p. or intranasal) with the presumed peripheral L‐DOPA decarboxylase inhibitor benserazide, but also each component alone can affect dopamine activity in the brain. Especially the findings with benserazide treatment might be of relevance for understanding the mechanisms of current L‐DOPA therapy, since they indicate that part of the treatments actions may possibly be determined by central dopaminergic effects of the accompanying amino acid decarboxylase inhibitor. Synapse 27:294–302, 1997.

Facilitation of memory by peripheral administration of substance P and naloxone using avoidance and habituation learning tasks

This paper summarizes results of a series of experiments dealing with the effects of the neuropeptide substance P (SP) on avoidance learning and habituation. Several doses of SP (0.5, 5, 50, 100, 250, 500 micrograms/kg) were administered posttrial intraperitoneally (IP). Three inhibitory one-trial avoidance tasks were used; uphill, step-down and step-through (alcove). Habituation was measured in an open field by recording the number of rearings. The posttrial injection of SP facilitated avoidance responses as well as reduced rearing in a dose- and time-dependent way. Pretraining and pretest injections (IP) of naloxone facilitated avoidance behavior and potentiated the action of SP, also in a dose-dependent manner. These results demonstrate that: a) peripheral posttraining administration of SP enhances memory; b) SP facilitates not only aversive or positively motivated learning tasks, but also habituation, which is a form of learning that involves neither positive nor negative reinforces; c) SP does not exert its effect by a long-lasting proactive action on performance during the testing trial; d) naloxone potentiates the SP posttraining effect. These data, therefore, suggest that memory-enhancing effects of SP are, at least in part, mediated via interactions between this peptide and endogenous opioid systems.

Effects on Anxiety and Memory of Systemic and Intra-Amygdala Injection of 5-HT3 Receptor Antagonist BRL 46470A

This study deals with the effects of the 5-HT3 receptor antagonist, BRL 46470A, on memory and anxiety, using the elevated T-maze. This method is useful for investigating the effects of anxiolytic drugs on memory, and the relationships between neural subsystems involved in emotionally related behaviors and in processes underlying learning. After the drug was either injected peripherally or microinjected into the amygdala, the animals were tested on the elevated T-maze (30 or 15 min later, respectively). Two kinds of aversively motivated behaviors, inhibitory avoidance and one-way escape, were recorded. These behaviors may reflect different types of fear/anxiety, namely, anticipatory anxiety and innate fear. Three days later, memory for these tasks was assessed by reexposing the subjects to the maze. The compound had an anxiolytic effect on the inhibitory avoidance response when given systemically, but an anxiogenic effect when injected into the amygdala. It had an anxiolytic action on the escape response when given either systemically or into the amygdala. The compound had no adverse effects on memory for either task. These results suggest that this new 5-HT3 antagonist may be useful in the treatment of certain types of anxiety disorders, especially those related to unconditioned fear, e.g. phobic or panic disorders, with the likelihood of having no side effects on memory processes. The contrasting results obtained with different measures of anxiety may also account for the inconsistencies found in the experimental literature dealing with compounds of this nature.

Facilitation of memory by peripheral administration of substance P

This report summarizes a recent series of experiments dealing with the effect of peripheral (i.p.) administration of SP on the learning of avoidance and habituation tasks. In summary, the results from these studies show that peripheral post-training SP administration in rats enhances memory in a dose- and time-dependent way. The effect of substance P on retention was observed across tasks with different response requirements and in the absence of explicit punishment. The memory-enhancing effects are long-lasting, until 21 days post-training, and are mediated, at least in part, via interactions with the endogenous opioid system. The mnemotropic effects of peripherally administered SP are sensitive to the functional integrity of the vagus, suggesting that the vagus nerve may be one pathway by which systemic SP influences memory storage processes in the brain. Furthermore, the data indicated that these effects seemed to be encoded by different SP sequences, the N-terminal SP1-7, but not the C-terminal hepta- and hexapeptide sequences being responsible for the memory-promoting effects. Taken together, these studies strongly suggest that SP may be considered to have memory-promoting effects.

Amnesia after diazepam infusion into basolateral but not central amygdala of Rattus norvegicus

Recent findings indicate that the memory-impairing effects of benzodiazepines may preferentially involve the basolateral nucleus of the amygdala. To test this hypothesis we examined the effects on pretrial injection of diazepam into the central as compared to the lateral/basolateral amygdaloid nuclei on memory for a conditioned avoidance response. Rats were implanted bilaterally with cannulae directed to either the central or lateral/basolateral amygdaloid nuclei. Five to 7 days later they were trained on a multitrial inhibitory avoidance (step-down) task to criterion and tested 48 h later. Fifteen minutes before training they were given an injection of either vehicle or diazepam (0.7 or 1.4 nmol) into the central or lateral/basolateral nuclei. Administration of diazepam into the lateral/basolateral nuclei but not the central nucleus induced anterograde amnesia. These results add to the body of data linking the GABA-benzodiazepine system of the lateral/basolateral nuclei to the amnestic effects induced by peripheral as well as central administration of benzodiazepines.