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The claustrum is a brain structure that remains shrouded in mystery due to the limited understanding of its cellular structure, neural pathways, functionality and physiological aspects. Significant research has unveiled connections spanning from the claustrum to the entire cortex as well as subcortical areas. This widespread connectivity has led to speculations of its role in integrating information from different brain regions, possibly contributing to processes such as attention, consciousness, learning and memory. Our working hypothesis posits that claustrum neural activity contributes to the acquisition, consolidation and reconsolidation of long-term memories in mice. We found evidence in CF-1 mice of a decline in behavioral performance in an inhibitory avoidance task due to intra-claustral administration of 2% lidocaine immediately after a training session or memory recall. Nevertheless, this does not seem to be the case for the acquisition or retrieval of this type of memory, although its neural activity is significantly increased after training, evaluated through c-Fos expression. Moreover, inhibition of the claustrum's synaptic activity appears to impair the consolidation but not acquisition or retrieval of an unconditioned memory formed in a nose-poke habituation task., (© 2024. The Author(s).)

Currently, testing for immunoglobulin E (IgE) sensitization is the cornerstone of diagnostic evaluation in suspected allergic conditions. This review provides a thorough and updated critical appraisal of the most frequently used diagnostic tests, both in vivo and in vitro . It discusses skin tests, challenges, and serological and cellular in vitro tests, and provides an overview of indications, advantages and disadvantages of each in conditions such as respiratory, food, venom, drug, and occupational allergy. Skin prick testing remains the first line approach in most instances; the added value of serum specific IgE to whole allergen extracts or components, as well as the role of basophil activation tests, is evaluated. Unproven, non-validated, diagnostic tests are also discussed. Throughout the review, the reader must bear in mind the relevance of differentiating between sensitization and allergy; the latter entails not only allergic sensitization, but also clinically relevant symptoms triggered by the culprit allergen., (© 2019 The Authors.)

Rationale: Parenteral injections of d-lysergic acid diethylamide (LSD), a serotonin 5-HT(2A) receptor agonist, enhance eyeblink conditioning. Another hallucinogen, (±)-1(2, 5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI), was shown to elicit a 5-HT(2A)-mediated behavior (head bobs) after injection into the hippocampus, a structure known to mediate trace eyeblink conditioning., Objective: This study aims to determine if parenteral injections of the hallucinogens LSD, d,l-2,5-dimethoxy-4-methylamphetamine, and 5-methoxy-dimethyltryptamine elicit the 5-HT(2A)-mediated behavior of head bobs and whether intrahippocampal injections of LSD would produce head bobs and enhance trace eyeblink conditioning., Materials and Methods: LSD was infused into the dorsal hippocampus just prior to each of eight conditioning sessions. One day after the last infusion of LSD, DOI was infused into the hippocampus to determine whether there had been a desensitization of the 5-HT(2A) receptor as measured by a decrease in DOI-elicited head bobs., Results: Acute parenteral or intrahippocampal LSD elicited a 5-HT(2A) but not a 5-HT(2C)-mediated behavior, and chronic administration enhanced conditioned responding relative to vehicle controls. Rabbits that had been chronically infused with 3 or 10 nmol per side of LSD during Pavlovian conditioning and then infused with DOI demonstrated a smaller increase in head bobs relative to controls., Conclusions: LSD produced its enhancement of Pavlovian conditioning through an effect on 5-HT(2A) receptors located in the dorsal hippocampus. The slight, short-lived enhancement of learning produced by LSD appears to be due to the development of desensitization of the 5-HT(2A) receptor within the hippocampus as a result of repeated administration of its agonist (LSD).

Previous studies have shown that amphetamine can enhance learning in Pavlovian conditioning tasks, but little is known about the changes in neural activity accompanying these performance enhancements. We evaluated the effects of amphetamine (10micromol/kg) on delay eyeblink conditioning performance and single-neuron activity in the anterior cingulate cortex (area 24) of the rabbit (Oryctolagus cuniculus). Amphetamine produced little to no learning enhancement on our task but strongly influenced the conditioned response (CR), which peaked closer in time to the onset of the unconditioned stimulus (US). The overall ACC population response showed very weak stimulus-evoked modulations during the course of training, with the primary effect being an increase in inhibition. Group discrepancies in stimulus-evoked inhibition correlated with differences in learning performance, and this correlation was stronger when subjects were grouped according to learning performance, independent of drug treatment. ACC neuronal responses of both groups displayed hemispheric asymmetries (laterality), but amphetamine treatment altered this effect, in that activity within each hemisphere of the amphetamine group more closely resembled that of the contralateral hemisphere of controls. Our data suggest that amphetamine modulates CR timing, and influences the flow of sensory information to the two cortical hemispheres. Our observations are also consistent with the ACC's non-essential role in learning during delay eyeblink conditioning.

Rationale: Associative learning during Pavlovian eyeblink conditioning has been shown to be regulated by 5-HT2A receptors. The existence of inverse agonists that retard learning through an action at the 5-HT2A receptor suggests the existence of constitutive activity at that receptor and that depletion of serotonin should have minimal effects on learning., Objectives: We examined whether depletion of serotonin would impair trace eyeblink conditioning or the enhancement of conditioning produced by the agonist lysergic acid diethylamide (LSD) and the retardation of conditioning produced by the inverse agonist MDL11,939., Methods: Animals received bilateral intraventricular injections of 5,7-dihydroxytryptamine (5,7-DHT) at doses of 760 or 1,140 microg/side (1.88 or 2.82 micromol/side) and were later exposed to eight daily conditioning sessions., Results: Serotonin depletion produced by the lower dose of 5,7-DHT was 71 and 72% in cortex and hippocampus, respectively, with no change in 5-HT2A receptor density, no effect on learning, and no effect on the ability of LSD to enhance and MDL11,939 to retard learning. The higher dose of 5,7-DHT produced serotonin decreases of 85 and 90% in cortex and hippocampus, respectively, accompanied by a 96% decrease in the density of the serotonin transporter, but no significant effect on learning., Conclusions: Pavlovian trace eyeblink conditioning is regulated predominantly by the constitutive activity of the 5-HT2A receptor rather than by serotonin release onto the receptor during learning. It was suggested that the 5-HT2A receptor regulates learning by modulating the release of dopamine, acetylcholine, and glutamate, transmitters known to affect eyeblink conditioning.

Serotonergic 5-HT2C and 5-HT1B receptors mediate inhibitory controls of eating. Questions have arisen about potential behavioral and neurological toxicity of drugs that stimulate the 2C site. We evaluated eating and other motor responses in male Dutch-belted rabbits after administration of m-chlorophenylpiperazine (mCPP). Studies conducted in vitro and in vivo assessed the pharmacological specificity of the ingestive actions of this agent. mCPP (0.15-10 micromol/kg sc) reduced consumption of chow and 20% sucrose solution with equal potencies (ED50 approximately equal 0.6 micromol/kg). In radioligand binding to rabbit cortex, mCPP displayed 15-fold higher affinity for 5-HT2C than for 5-HT1B receptors. The serotonin antagonist mesulergine (7000-fold selective for 5-HT2C) reversed the hypophagic action of mCPP, but the 5-HT1B/1D antagonist GR127,935 did not. GR127,935 (0.5 micromol/kg) did prevent hypophagia produced by the highly selective 5-HT1B/1D agonist GR46,611. Observational methods demonstrated that mCPP decreased the frequency of eating chow but increased other motor activities. When rabbits consumed sucrose, videoanalysis revealed that mCPP reduced total time licking and the duration of individual bouts, but not bout frequency or the actual rate of consumption. mCPP increased locomotor and other activities, and greatly increased vacuous oromotor stereotypies and tongue protrusions. Nonetheless, rabbits licked accurately at the spout for sucrose. When sucrose was infused intraorally through a cheek catheter, mCPP actually increased the peak amplitude and overall magnitude of jaw movements. We conclude that mCPP stimulates 5-HT2C receptors to reduce food intake in rabbits. This hypophagia involves disruption of appetitive components of eating and is accompanied by adverse motor actions. This profile raises questions about the use of the 5-HT2C receptor as a target for novel therapeutic agents for obesity.

Rationale: Associative learning during classical trace eyeblink conditioning has been shown to be regulated by serotonin 5-HT(2A )receptors and to be critically dependent on the integrity of frontal cortex. Chronic administration of 5-HT(2A) ligands has been shown to produce a selective up- or down-regulation of 5-HT(2A) receptors in frontal cortex., Objectives: We examined whether alterations in 5-HT(2A) receptor density had a functional significance with respect to associative learning., Methods: Animals received chronic injections of LSD, BOL or MDL11,939 and 1 day later began classical trace conditioning of the eyeblink response., Results: The density of 5-HT(2A) receptors in frontal cortex was significantly increased at 1-4 days after the cessation of chronic injections of the selective 5-HT(2A) receptor ligand MDL11,939. Rabbits demonstrated an enhancement of associative learning when training began at 1 day after cessation of chronic MDL11,939 injections, but acquired at the same rate as controls when training began at 8 days after cessation of injections, a time when receptor density had returned to control levels. Animals that began training 1 day after chronic injections of BOL or LSD, drugs that produce decreases in 5-HT(2A) receptor density, demonstrated normal rates of acquisition., Conclusions: These results indicate that increases in the density of 5-HT(2A) receptors in frontal cortex are associated with increases in the rate of associative learning, and further support an important role for this receptor in cortical circuitry that mediates learning. More generally, these results suggest an approach for functional remodeling of brain regions in the adult animal.

This study examined the role of the mitogen-activated protein kinase (MAPK) family during acquisition of the rabbit's classically conditioned eye-blink response. Eye-blink conditioning produced a significant, bilateral activation of both extracellular signal-regulated protein kinases (ERKs) and p38 MAPK in the anterior cerebellar vermis. There was also a significant bilateral activation of ERKs in the dorsal hippocampus with no change in p38 MAPK. These changes were seen at 2 min after the last conditioning session, were maintained for at least 180 min, and occurred without any change in the protein expression of either ERKs or p38 MAPK. There were no changes in ERKs or p38 MAPK in frontal cortex, in cerebellar hemispheral lobule VI, or in a section of brainstem containing the inferior olive. Moreover, the stress-related protein kinase Jun N-terminal kinase (JNK), another subfamily of MAPKs, was not altered in any of the brain regions examined. Animals receiving explicitly unpaired presentations of a conditioned stimulus and an unconditioned stimulus did not acquire conditioned responses (CRs) and did not demonstrate any changes in ERKs, p38 MAPK, or JNK. The intraventricular injection of SB203580, a selective p38 MAPK inhibitor, significantly retarded CR acquisition and blocked the learning-related increases in p38 MAPK activity in the anterior vermis. PD98059, a selective MAPK kinase inhibitor, had a smaller and only marginally significant effect on CR acquisition, although it did block the learning-related increases in ERK activity in both the hippocampus and anterior vermis. These results indicate that p38 MAPK is activated during associative learning and may play a role in the transcriptional events that lead to memory consolidation.

Earlier studies of human infants and studies employing animal models had indicated that prenatal exposure to cocaine produced developmental changes in the behavior of the offspring. The present paper reports on the results obtained in a rabbit model of in utero exposure to cocaine using intravenous injections (4 mg/kg, twice daily) that mimic the pharmacokinetics of crack cocaine in humans. At this dose, cocaine had no effect on the body weight gain of dams, time to delivery, litter size and body weight or other physical characteristics of the offspring. In spite of an otherwise normal appearance, cocaine-exposed neonates displayed a permanent impairment in signal transduction via the D1 dopamine receptor in caudate nucleus, frontal cortex and cingulate cortex due to an uncoupling of the receptor from its associated Gs protein. This uncoupling in the caudate nucleus was shown to have behavioral consequences in that young or adult rabbits, exposed to cocaine in utero, failed to demonstrate amphetamine-elicited motor responses normally seen after activation of D1 receptors in the caudate. The cocaine progeny also demonstrated permanent morphological abnormalities in the anterior cingulate cortex due to uncoupling of the D1 receptor and the consequent inability of dopamine to regulate neurite outgrowth during neuronal development. Consistent with the known functions of the anterior cingulate cortex, adult cocaine progeny demonstrated deficits in attentional processes. This was reflected by impairment in discrimination learning during classical conditioning that was due to an inability to ignore salient stimuli even when these were not relevant to the task. The impairment in discrimination learning also occurred in an instrumental avoidance task and could be shown to be due to an impairment of cingulothalamic learning-related neuronal coding. It was proposed that the selective loss of D1-related neurotransmission in the anterior cingulate cortex prevented an appropriate activation of GABA neurons and thus a loss of inhibitory regulation that is necessary for processes involved in associative attention. Taken together, these findings suggest that the uncoupling of the D1 receptor from its G protein may be the fundamental source of the anatomic, cognitive and motor disturbances seen in rabbits exposed to cocaine in utero. Moreover, the long-term cognitive and motor deficits observed in the rabbit model are in agreement with the recent reports indicating that persistent attentional and other behavioral deficits may be evident in cocaine-exposed children as they grow older and are challenged to master more complex cognitive tasks.

Serotonin 5-HT(2) antagonists that significantly retard the acquisition of classically conditioned responses (CRs) also impair the performance of the unconditioned response (UR). Effects on the UR appear to be due to an inverse agonist action at the 5-HT(2) receptor. These findings raised the possibility that the learning deficits were either secondary to a performance deficit and/or that the retardation of learning was not due to actions at the serotonin 5-HT(2) receptor. In this study, we examined the effects of the 5-HT(2)-receptor antagonists, namely mianserin and D-2-bromolysergic acid diethylamide hydrogen tartrate (BOL), on CR acquisition. We also determined whether the retarded acquisition of CRs produced by mianserin (a) was due to an action at the 5-HT(2) receptor and (b) was secondary to a performance deficit. Effects of drugs on CR acquisition, maintenance, and retention were determined during trace-conditioning of the rabbit's nictitating membrane (NM) response. BOL (0.058 to 5.8 micromol/kg) had no effect on CR acquisition, whereas mianserin (0.1 to 10 micromol/kg) produced a significant and dose-dependent retardation of CR acquisition. The retarded CR acquisition produced by mianserin (10 micromol/kg) was due to its actions at the 5-HT(2) receptor, because this effect was completely blocked by a dose of BOL (5.8 micromol/kg) that had no effect when given alone. Neither maintenance nor retention of learning was affected by mianserin treatment during acquisition. We conclude that mianserin acts as an inverse agonist at the serotonin 5-HT(2) receptor to produce both a retardation of CR acquisition and an impairment of the UR. However, the learning and performance effects of mianserin are separable.

Hippocampal multiple unit activity was recorded in rabbits during each of four preacquisition treatments and during subsequent classical conditioning of the nictitating membrane response. Three preexposure conditions were employed: CS alone presentations, presentations of the CS paired with a second, neutral stimulus, or unpaired presentations of the CS and second stimulus. It was predicted that (a) CS alone preexposures would produce a decrease in hippocampal activity and a retarded rate of subsequent conditioned response (CR) acquisition and (b) the magnitude of both effects would be attenuated by preexposures of the CS paired with a second stimulus. The results partially supported both predictions. Hippocampal activity was inhibited during CS alone preexposures and that inhibition was attenuated by pairing the CS with a second, neutral stimulus. Behaviorally, all of the preexposure groups showed equivalent, retarded rates of acquisition compared to a nonpreexposed control group. Hippocampal activity throughout acquisition was significantly greater in the nonpreexposed group compared to the group preexposed to the CS alone. Hippocampal activity of the other two groups was intermediate between the nonpreexposed and the CS alone groups. It is suggested that alterations in the magnitude of hippocampal activity may provide a reliable, neuronal correlate of CS associability changes.

This study examined the role of the serotonin 5-HT2 receptor in motor function by examining the effect of antagonists on the motor performance of a cranial nerve reflex, the nictitating membrane (NM) reflex of the rabbit. The NM reflex was elicited by varying intensities of a tactile stimulus and the magnitudes of the elicited responses were measured at each intensity. Dose-response curves were obtained for the effects of several 5-HT2 receptor antagonists on response magnitude. d-Bro-molysergic acid diethylamide (BOL), LY-53,857 and ketanserin had no significant effect on the magnitude of the NM reflex, indicating that they are neutral antagonists. However, the 5-HT2 receptor antagonists ritanserin, MDL-11,939 and mianserin produced a significant reduction in response magnitude with no significant effects on response frequency, suggesting that they were acting as inverse agonists at the 5-HT2 receptor. The reduction in reflex magnitude produced by mianserin (10 micromol/kg) was fully blocked by BOL (5.8 micromol/kg), supporting the conclusion that mianserin was producing a reduction in reflex magnitude through an effect at the 5-HT2 receptor. The occurrence of inverse agonism suggests the possible existence of constitutive activity in vivo. We conclude that the 5-HT2 receptor (either 2A or 2C) plays an important role in motor function, perhaps by providing a tonic influence on motor systems.

We have developed a rabbit model of in utero exposure to intravenous injections of cocaine given twice daily to dams from gestational days 8-29. At the doses employed (4 mg/kg, injected twice daily), no differences were found in the body weight gain of dams, time to delivery, litter size, and body weight or other physical characteristics of the offspring. However, cocaine-exposed pups displayed an abnormal structural and neurochemical development of the anterior cingulate cortex which persisted into adulthood. In agreement with the known functions of the anterior cingulate cortex, we found that adult, sexually mature rabbits, exposed to cocaine prenatally, demonstrate impairments in motor function, alterations in associative learning and severe impairments in discrimination learning. Moreover, the alterations in discrimination learning were interpreted to be due to deficits in attentional processes. Specifically, cocaine progeny preferentially attend to more salient stimuli even when these are not relevant to the task. Consequently they have difficulty in attending to less salient but relevant stimuli when more salient but irrelevant stimuli occur in the same context. We concluded that the learning deficits are a reflection of the morphologic and neurochemical abnormalities of the anterior cingulate cortex. Alterations in dopamine function of the caudate nucleus may also contribute to the deficits in motor performance.

We have developed a rabbit model of in utero exposure to intravenous injections of cocaine given twice daily to dams from gestational days 8-29. At the doses employed (4 mg/kg, injected twice daily), no differences were found in the body weight gain of dams, time to delivery, litter size, and body weight or other physical characteristics of the offspring. However, cocaine-exposed pups displayed an abnormal structural and neurochemical development of the anterior cingulate cortex which persisted into adulthood. In agreement with the known functions of the anterior cingulate cortex, we found that adult, sexually mature rabbits, exposed to cocaine prenatally, demonstrate impairments in motor function, alterations in associative learning and severe impairments in discrimination learning. Moreover, the alterations in discrimination learning were interpreted to be due to deficits in attentional processes. Specifically, cocaine progeny preferentially attend to more salient stimuli even when these are not relevant to the task. Consequently they have difficulty in attending to less salient but relevant stimuli when more salient but irrelevant stimuli occur in the same context. We concluded that the learning deficits are a reflection of the morphologic and neurochemical abnormalities of the anterior cingulate cortex. Alterations in dopamine function of the caudate nucleus may also contribute to the deficits in motor performance.

The 5-HT(2A/2C) receptor antagonist, ritanserin, was reported to retard the acquisition of conditioned responses (CRs) during classical conditioning of the rabbit's nictitating membrane (NM) response. The present study compared the effects of ritanserin on acquisition of CRs to a tone conditioned stimulus (CS) with that of the 5-HT(2A/2C) receptor antagonist, LY-53,857 and the 5-HT2A selective antagonist, MDL-11,939. All three drugs were injected at equimolar doses of 0.067, 0.67 and 6.7 micromol/kg, SC, 1 h before behavioral testing. Ritanserin and MDL-11,939 retarded CR acquisition to a tone CS, while LY-53,857 had no effect. Control experiments demonstrated that ritanserin (1 micromol/kg), MDL-11,939 (1 micromol/kg) and LY-53,857 (2 micromol/kg) had no effect on baseline responding or non-associative responding to the CS. However, both ritanserin and MDL-11,939 impaired the performance of the unconditioned NM reflex, as measured by a decrease in UR amplitudes on US alone trials, while LY-53,857 had no effect. In previously trained animals, ritanserin robustly impaired the performance of CRs, as measured by a reduced ability of the CS to elicit CRs, while the effects of LY-53,857 and MDL-11,939 were marginal. The retardation of associative learning produced by ritanserin and MDL-11,939 may have been due, at least in part, to their impairment of the NM reflex arc. Since MDL-11,939 is a highly selective 5-HT2A antagonist, the retardation of learning and impairment of UR amplitudes produced by MDL-11,939 and ritanserin may have been due to blockade of the 5-HT2A receptor. The ability of ritanserin and MDL-11,939 to produce effects on learning and performance that were opposite to that of 5-HT(2A/2C) agonists suggests that they may be acting as inverse agonists at that receptor. These results stress the importance of the serotonergic system for optimal associative learning and motor function.

d-Lysergic acid diethylamide (LSD), an agonist at the 5-HT(2A/2C) and 5-HT1A receptors, has previously been demonstrated to enhance associative learning as measured by accelerated acquisition of the rabbit's classically conditioned nictitating membrane (NM) response. The present study examined further the role of these receptors in the action of LSD. LSD (30 nmol/kg, I.V.) significantly enhanced conditioned response (CR) acquisition to both tone and light conditioned stimuli (CSs), while the 5-HT1A receptor agonists 8-hydroxy-2-(dipropylamino)tetralin (8-OH-DPAT; 50 and 200 nmol/kg) and lisuride (0.3-30 nmol/kg) had no effect. Ritanserin (6.7-6700 nmol/kg, S.C.), a selective 5-HT(2A/2C) receptor antagonist, retarded acquisition of CRs to both tone and light CSs in a dose-dependent manner. Ritanserin (6.7-670 nmol/kg, S.C.) also dose dependently antagonized the enhancement of CR conditioning produced by LSD (30 nmol/kg, I.V.) to both tone and light CSs. We conclude that the enhancement of CR acquisition by LSD was due to an action at the 5-HT(2A/2C) receptor. These results suggest that the 5-HT(2A/2C) receptor plays an important role in learning.

The nictitating membrane (NM) reflex was assessed in adult Dutch-belted rabbits exposed to cocaine in utero. The intensity threshold for eliciting the reflex was increased in cocaine progeny and the amplitude of the reflex was decreased at the lower stimulus intensities. However, cocaine and saline progeny showed equivalent rates of habituation of the NM reflex when tested with a suprathreshold eliciting stimulus. Reliable modification of the NM reflex was obtained when the reflex-eliciting stimulus was preceded by an auditory stimulus at intervals of 100-800 ms. Cocaine and saline progeny exhibited an increase in the peak amplitude of the reflex, a shortening of the latency of the reflex, and a shortening of the latency to achieve peak amplitude of the reflex as a function of increases in the interstimulus interval. Furthermore, cocaine progeny showed significantly longer response latencies than saline progeny across all interstimulus intervals, although neither the peak amplitude nor the latency to achieve peak amplitude was affected. Thus, prenatal exposure to cocaine affected elicitation of the defensive NM reflex to an aversive stimulus but did not affect the sensorimotor integration necessary for modification of the reflex by antecedent stimulation.

Previous studies had shown that intrauterine exposure to cocaine produces an increase in the number of immunoreactive GABA neurons and abnormal dendritic structure of pyramidal cells in the anterior cingulate cortex, a brain region known to be involved in attentional processes and discrimination learning. Because structural abnormalities might be expected to produce related functional deficits, we examined whether intrauterine exposure to cocaine would affect discrimination learning in adult rabbits. We previously reported that cocaine progeny undergoing concurrent acquisition to visual and auditory CSs show a normal rate of learning to a light CS and an accelerated rate of learning to a tone. Here, we report that adult, Dutch-belted rabbits exposed to cocaine in utero showed impaired discrimination learning when responding to a positive visual cue but not when responding to a positive auditory cue. The nature of the deficit consisted of an impaired ability to acquire learned responses to the visual CS+ rather than in an impaired ability to withhold responses to the auditory CS-. Given that auditory stimuli tend to be more salient than visual stimuli in the normal rabbit, the preceding pattern of results suggests that intrauterine cocaine exposure affected the ability to preferentially attend to less salient but relevant stimuli and to ignore more salient, irrelevant stimuli. More importantly, these results indicate that prenatal exposure to cocaine produces neurobehavioral abnormalities which persist into adult life.

This study examined the effects of hypotensive stress on classical conditioning of the rabbit's nictitating membrane response. Hypotension, consisting of an approximately 45% decrease in blood pressure, was maintained for 30 min by the i.v. infusion of sodium nitroprusside. Twenty minutes later animals were exposed to a conditioning session consisting of 60 pairings of a 200 ms tone conditioned stimulus with a 100 ms airpuff unconditioned stimulus directed at the cornea. This procedure was repeated for four consecutive days. Animals exposed to the hypotensive stress demonstrated a significantly retarded acquisition of conditioned responses as measured by their frequency and onset latency as well as by an increase in the number of trials required to reach acquisition criteria of five and 10 consecutive conditioned responses as compared with controls. A separate group of animals received a nitroprusside infusion one day after the acquisition of conditioned responses to the tone conditioned stimulus. These animals demonstrated a normal retention of conditioned responses and a normal response to varying intensities of the conditioned stimulus. Hypotensive stress also had no effect on the frequency and topography of the unconditioned response. It was concluded that a decrease in blood pressure can serve as a physiological stressor. One of the reactions to this stress consists of a retardation in the formation of associations during a learning task, without any decrease in the ability to retrieve previously learned material.

Previous studies had demonstrated that in utero exposure to cocaine produces structural changes in the development of the rabbit's anterior cingulate cortex. Because the anterior cingulate cortex has been proposed to subserve a variety of cognitive processes including associative learning, we investigated the effects of intrauterine exposure to cocaine on the acquisition of the rabbit's classically conditioned nictitating membrane response. Adult, sexually mature rabbits born of dams that had received intravenous injections of either saline or cocaine (4 mg/kg, twice a day) from day 8 to day 29 of gestation were classically conditioned by pairing tone and light CSs with an airpuff US. Rabbits that had been exposed to cocaine in utero demonstrated a more rapid acquisition of CRs to a tone CS but not to a light CS as compared with saline controls. Control experiments indicated that the accelerated learning to the tone CS was not due to sensitization, pseudoconditioning, altered baseline rate of responding, an increased responsiveness to the airpuff US, or to a change in the intensity threshold of the tone CS for elicitation of CRs. We conclude that in utero exposure to cocaine alters the processing of auditory stimuli and this leads to an abnormally rapid acquisition of CRs. It is suggested that this functional consequence of prenatal exposure to cocaine is due to structural abnormalities in anterior cingulate cortex.

The rabbit has been used to model the potential effects of in utero exposure to cocaine on fetal and postnatal development. Special advantages of this animal model include the fact that cocaine can be easily administered intravenously, thus mimicking crack cocaine use by pregnant women. Results indicate that at the dosage used (8 mg/kg of body weight, given intravenously daily) gross teratologic defects do not develop. Cocaine-exposed pregnant does not differ from controls in weight gain or in the number of live kits delivered. Cocaine-exposed kits do not differ from controls in survival or in postnatal weight gain. The importance of this rabbit model is that offspring that have been exposed to these doses of cocaine in utero have a variety of abnormalities in structure and function of the central nervous system in the absence of any major teratologic defects.

This study examined the effects of the hallucinogen, MDA, on brain content of monoamines and their metabolites in the rabbit. A single 1.8 mg/kg dose of MDA produced 30 to 64% increases in the 5-HT content of frontal cortex from 30 to 120 min after injection and a decrease in 5-HT turnover from 30 min to 8 h, but had no effect in hippocampus, caudate nucleus, or hypothalamus. A single 3.6 mg/kg dose of MDA also reduced the turnover of 5-HT in frontal cortex, but this was accompanied by a decrease in 5-HIAA with no increase in 5-HT. The 1.8 and 3.6 mg/kg doses of MDA had no significant or consistent effects on the contents of DA, DOPAC, HVA, and NE in any brain area examined. Chronic administration of MDA (3.6 mg/kg/day for 4 days) failed to produce any evidence of a neurotoxic action on 5-HT neurons. Higher doses could not be employed because the LD50 of MDA was approximately 5 mg/kg. This study has demonstrated that behaviorally effective and nonneurotoxic doses of MDA produce increases in the content and decreases in turnover of 5-HT in frontal cortex that resemble those of other hallucinogens such as LSD and DOM.

The rate of associative learning was assessed in the presence of saline versus methylenedioxymethamphetamine (MDMA) at doses of 0.95, 1.9, and 3.8 mg/kg. The conditioned stimuli (CSs) were lights and tones and the unconditioned stimulus (US) was a corneal air puff. Learning was enhanced by all but the highest dose of drug tested, and the enhancement was most pronounced when light was used as the conditioned stimulus. Nonassociative responding was assessed using unpaired presentations of the lights, tones, and air puffs. MDMA (1.9 mg/kg) produced a slight increase in the percentage of baseline responses but failed to produce an increase in the frequency of nonassociative responding in the presence of the lights or tones. MDMA produced a significant increase in the amplitude of the unconditioned response to the corneal air puff across the 10 sessions. This increase was taken as evidence for sensitization of the unconditioned response, a nonassociative learning phenomenon. In summary, MDMA, like the parent compound methylenedioxyamphetamine (MDA), enhances both conditioned and unconditioned responding. Because this dual effect has not been seen with related psychedelic compounds, the effect appears to be unique to this class of phenylethylamine drugs.

This study employed immunocytochemistry to visualize the neurotoxic effects of methylenedioxyamphetamine hydrochloride (MDA) on serotonergic projections to brainstem structures. Separate groups of animals were injected twice a day, for 4 consecutive days, with: saline; MDA (40 mg/kg/day); or fluoxetine hydrochloride (10 mg/kg) prior to each injection of MDA. In agreement with previous reports, MDA produced a pronounced loss of 5-HT immunoreactivity in the forebrain, most notably in neocortex and hippocampus. However, our results revealed that MDA also produced a loss of 5-HT fibers in brainstem that was as severe as that seen in any region of forebrain. Regions most severely affected included: superior colliculus; superior olivary complex; trigeminal sensory complex and vestibular nuclei. The brains of animals treated with MDA demonstrated a relative absence of fine 5-HT axon terminals within these forebrain and brainstem regions, while thicker axonal elements were still present. The neurotoxic effects of MDA on serotonergic projections to forebrain and brainstem were completely blocked by the prior administration of the 5-HT reuptake inhibitor, fluoxetine. It was suggested that the denervation of the superior colliculus, superior olive and vestibular nuclei could alter visually guided eye movements and the vestibulo-ocular reflex while the loss of serotonergic inputs to the trigeminal sensory complex might be expected to alter tactual reflexes.

Pavlovian conditioning of the rabbit's nictitating membrane response was used to determine whether the retardation of learning produced by enkephalin derivatives was mediated through an action on mu- or delta-opioid receptors. Intraventricular injection of the highly selective mu-agonist, [D-Ala2, MePhe4, Gly-ol5] enkephalin (DAMGO) significantly retarded (2 nmol) or completely blocked (20 nmol) the acquisition of conditioned responses. The retarded or blocked acquisition of conditioned responses produced by DAMGO could still be detected when the rabbits were tested two days after cessation of drug injections, suggesting that DAMGO had affected learning and not simply performance of conditioned responses. Intraventricular injection of a comparable dose (20 nmol) of the highly selective delta-agonist, [D-Pen2,5] enkephalin (DPDPE) did not retard learning. A higher dose of DPDPE (200 nmol), which would be expected to act at both mu- and delta-receptors, did produce a retardation of acquisition that was comparable to the effects of a 2-nmol dose of DAMGO. However, unlike DAMGO, the retardation of learning produced by this dose of DPDPE appeared to be due, at least in part, to a reduction in the aversive properties of the unconditioned stimulus. The retardation of learning produced by 2 nmol of DAMGO and 200 nmol of DPDPE was due to an action on an opioid receptor because this effect was blocked by a subcutaneous injection of naloxone (2.5 mumol/kg), a specific opioid antagonist. The stereoselective effects of the mu-opioid agonist levorphanol further confirmed that the retarded learning was mediated through actions at the mu-opioid receptor. Autoradiographic studies indicated that a 2-nmol, intraventricular dose of DAMGO achieved contents greater than 1 pmol/mg in tissue lying within 2 mm of the lateral ventricle, third ventricle and cerebral aqueduct. These data suggest that enkephalins produce a retardation of learning through an effect on the mu-type of opioid receptor and that at least one locus of their actions is produced at periventricular sites lying within 2 mm of the ventricles.(ABSTRACT TRUNCATED AT 250 WORDS)

The behavioral effects of a single, acute dose of methylenedioxyamphetamine (MDA) were assessed using the rabbit nictitating membrane (NM) preparation. Acquisition of the classically conditioned NM response was carried out in a single session under MDA at doses of 10 and 20 mumol/kg (1.79 and 3.58 mg/kg, respectively) and retention was assessed during a drug-free state 48 h later. In agreement with previous reports, MDA at a dose of 10 mumol/kg enhanced acquisition of the conditioned NM response and learning was retained in the nondrug state. However, MDA at a dose of 20 mumol/kg failed to affect either acquisition or retention. In further agreement with previous reports, MDA at 10 mumol/kg facilitated the amplitude of the unconditioned response during unpaired stimulus presentations. It is suggested that this enhanced motor response may be due to enhanced neural transmission in the afferent limb of the unconditioned reflex arc.

This study reexamined the effects of unilateral damage to cerebellar hemispheral lobule VI on the rabbit's conditioned nictitating membrane (NM) response. Extensive unilateral removal of hemispheral lobule VI in 11 rabbits impaired ipsilateral conditioned responses as reflected by reductions of 52% in mean frequency and 53% in mean amplitude during test trials on the first postoperative session. The decreases in the amplitude and frequency of conditioned responses were highly correlated (r = 0.82). The frequency of conditioned responses recovered to control levels but their amplitudes remained reduced such that the correlation between these two measures of responding was no longer significant by the 12th postoperative conditioning session. The decrease in the amplitude of conditioned responses was not accompanied by changes in onset latency or rise time. There was no significant impairment of conditioned responses in surgical controls and animals with only partial damage to hemispheral lobule VI. It was concluded that hemispheral lobule VI plays an important role in the regulation of motor centers in the brainstem so as to facilitate the initiation and optimum execution of the conditioned NM reflex. This cortical regulation of the conditioned NM response may contain learned elements; however, these cannot be resolved with lesion methods, nor has their existence been proven in this or other lesion studies. Nevertheless, the results of this study do demonstrate that the cerebellar cortex cannot be considered as the single locus necessary for NM conditioning.

In this study we examined the effects of harmaline on Pavlovian conditioning of the rabbit's nictitating membrane response. The acquisition of conditioned responses was determined during a single session consisting of 120 pairings of a tone-conditioned stimulus with a corneal air puff unconditioned stimulus. Harmaline severely retarded (5 mg/kg) or completely blocked (10 and 20 mg/kg) acquisition of conditioned responses. The blocked or retarded acquisition of conditioned responses could still be detected when the rabbits were tested 2 d after cessation of drug injections, suggesting that harmaline was affecting acquisition and not the motoric expression of conditioned responses. Control experiments established that harmaline (5 mg/kg) did not affect (1) baseline levels of responding, (2) the level of non-associative responding to the conditioned stimulus, (3) the amplitude or any of the temporal characteristics of the unconditioned response, (4) the development of habituation to the unconditioned stimulus, and (5) the threshold of the unconditioned stimulus for eliciting the unconditioned response. However, harmaline did produce a 12 dB increase in the intensity threshold of the conditioned stimulus for eliciting conditioned responses. We concluded that the primary effect of harmaline was to impair stimulus processing within brainstem circuits such as to reduce the excitatory properties of the conditioned stimulus, thus retarding its entry into associative learning. The results were discussed with respect to the possible role of the inferior olive in associative learning.

The effects of methylenedioxyamphetamine (MDA) on classical conditioning of the rabbit's nictitating membrane response were assessed in four experiments. Experiment 1 established a dose-effect curve for MDA doses of 0, 1, 3, and 10µmol/kg. Both the 3 and 10µmol/kg doses significantly enhanced the rate of learning. Experiment 2 established that the 10µmol/kg dose of MDA had no effect on non-associative determinants of responding to the conditioned stimulus (CS). However, this dose of MDA apparently sensitized the response to the unconditioned stimulus (US). Experiments 3 and 4 assessed the effects of the 10µmol/kg dose of MDA on sensory processing of the CS and US, respectively. MDA had no effect on the intensity thresholds for eliciting responses to either of these stimuli. Thus, the enhanced rate of acquisition observed in experiment 1 cannot be attributed to an increase in nonassociative responding to the CS or to enhanced sensory processing of the CS or US. MDA apparently enhanced the rate of acquisition in experiment 1 by facilitating the association between the CS and US. These findings are unique in that no other hallucinogen we have examined has enhanced acquisition without also affecting either non-associative responding or sensory processing of the CS and US.

Four studies were conducted to delineate potential neural processes involved in retention of a peripherally induced postural asymmetry. In the first experiment, dorsal and ventral spinal root section following varying intervals of stimulation successfully abolished peripherally induced hindlimb asymmetry. Experiment 2 revealed that 50 min of ventral root stimulation, in the absence of central connection, was not effective in producing asymmetry persistence. In an attempt to more closely delineate the time parameters involved in peripherally induced asymmetry retention, Experiment 3 was conducted. Asymmetry was found to consistently outlast a spinal transection if 40 min of stimulation was given. Finally, possible modulatory higher brain center influences on the retention processes were demonstrated in Experiment 4. Asymmetry persistence was consistently observed in animals that received 10, 20, 30, 40, or 50 min of hindlimb stimulation if 50 min were allowed to elapse between the onset of stimulation and spinal section. These results, when coupled with the findings of earlier studies, suggest an active involvement of spinal reflex centers in the fixation process. In addition, these studies indicate that the manner in which reflex activity is altered is not crucial as long as underlying time parameters are adhered to. Finally, the present studies demonstrate that higher center influences can modulate retention of a postural asymmetry in a complex manner.

The effects of dorsal hippocampal lesions on retention of classical trace conditioned responses were examined using the rabbit nictitating membrane preparation. Animals were trained to criteria and then lesioned either in the cortex or in the hippocampus and the cortex. Hippocampal damage had no effect on the retention of responses but produced significantly longer onset latencies. A control group of hippocampal animals acquired conditioned responses (CRs) at least as quickly as the prelesion subjects, and they also exhibited longer response onset latency. A second experiment evaluated the performance of hippocampal lesioned animals in classical trace conditioning with either a low-intensity periorbital shock or corneal air puff as the unconditioned stimulus (UCS). Hippocampal animals successfully acquired CRs under both conditions but exhibited an alteration of response onset which was dependent on the form of the UCS. Hippocampal animals displayed shorter response onset in the air-puff condition and longer response onset in the shock condition. Cortical animals timed responses consistently regardless of the UCS. These findings strongly suggest that the hippocampus modulates temporal characteristics of learned behavior.

These experiments were conducted to examine extinction and retention of a classically conditioned response in acute spinal cats. Conditioning was obtained by pairing a mild electrical stimulus to the superficial peroneal sensory nerve (CS) with a stronger electrical stimulus to the ankle skin (US) of the same leg. Subsequent extinction was produced by presenting CS-alone trials. In the first experiment, animals given massed extinction trials showed response decrements to base levels. Response decrements were not found in animals that received distributed extinction trials. In the second experiment, .5-, 1-, 2-, 3-, or 4-hr intervals between acquisition and extinction produced no significant differences in the extinction data. In the third experiment, animals received extinction trials immediately or 30 min after acquisition trials, followed by 20 additional extinction trials 30 min later. Analyses of these data indicated significant acquisition and extinction in the 10- and 20- acquisition trial groups. As in Experiment 2, varying the interval between acquisition and extinction did not produce any group differences in the extinction data. The results of these experiments demonstrate that response increases produced by paired trials in the spinal preparation do not decay spontaneously over time and are not caused by sensitization effects.

Two methods were used to demonstrate retention of postural asymmetries after spinal cord section in adult rats. In the first preparation, postural asymmetries of the hindlimbs were induced by placing electrolytic lesions in the anterior cerebellum. Asymmetry was found to consistently outlast a spinal cord section if 45 min were allowed between brain lesion and cord section. A certain percentage of rats allowed 35 or 40 min also demonstrated the retention. In the second preparation, postural asymmetries induced by 45 min of direct hindlimb stimulation were also retained after spinal section. Rhizotomy prior to stimulation resulted in a lack of appreciable asymmetry upon termination of the stimulation. Finally, retention of a hindlimb-stimulation-induced asymmetry was observed in animals that underwent a spinal section prior to the stimulation. The present series of studies demonstrates that the "spinal fixation" phenomenon can be obtained by induction of postural alterations from central (cerebellar lesion) and peripheral (hindlimb stimulation) sources. In addition, results obtained from spinal animals indicate that retention of peripherally induced asymmetry is not crucially dependent on higher brain center activity but rather seems to be more dependent on long-term alterations that occur directly in the spinal reflex system.

Temporal parameters concerning retention of a stimulation-induced hindlimb flexion were investigated. After spinal transection, 60 rats received 10, 20, 30, 40, 45, or 50 min of hindlimb stimulation. Consistent persistence of flexion following stimulus cessation was observed in rats that were stimulated for at least 30 min. When the results ae compared with previous data, the spinalized rat shows a 10-min reduction in the critical time interval needed to consistently obtain poststimulation persistence of flexion.

The nictitating membrane response of rabbits was conditioned at a 200 msec interstimulus interval (ISI) with either circumorbital (C) or paraorbital (P) shock as the unconditional-stimulus locus. After 3 acquisition days half of each group was shifted to a 700 msec interstimuls interval. Results indicated: (1) more rapid acquisition for Group C, (2) postshift response decrements for both groups, (3) more rapid and stable, as well as complete return to preshift performance levels for Group C. Results were discussed in terms of the response-shaping hypothesis and the contiguity-substitution hypothesis in explaining both conditional response emergence and subsequent modifications of CR topography.