Repeated Acquisition Task Research Articles

Behavioral and Molecular Effects of THC and CBD in Adolescent Nonhuman Primates

The marijuana plant produces over 100 different cannabinoids, including Δ9− tetrahydrocannabinol (THC) and cannabidiol (CBD). THC concentrations in retail marijuana have risen dramatically, while CBD levels have declined. High concentrations of THC and high ratios of THC:CBD in marijuana are thought to be associated with more robust psychoactive effects, while CBD has been reported to attenuate THC‐induced effects. The current study sought to compare the behavioral and molecular effects of THC alone and THC combined with CBD in adolescent nonhuman primates in order to investigate, across a wide range of parameters, whether CBD modulates the effects of THC.Twelve male adolescent squirrel monkeys (2.3 – 2.5 years) were divided into three treatment groups: vehicle control, THC, or THC+CBD (n=4/group). Initially they were treated with low doses of THC or THC+CBD weekly for four weeks. Thereafter, at the onset of the testing regimen, animals received THC (1 mg/kg) or THC+CBD (1 mg/kg+3mg/kg) daily for 4 months. The following parameters were measured: 1. Cognitive ability: Animals were trained to engage in touchscreen‐based cognitive tasks according to previously established methods from our laboratory. 2. Observed behavior: behavior was video‐taped weekly in a Plexiglass observation chamber and rated by an observer blinded to the drug regimen. 3. Blood THC, CBD, and THC metabolites were measured four times over the course of the experiment. THC levels in two brain regions were measured following euthanasia. 4. mRNA levels in selected brain regions were assessed by bulk RNAseq and confirmed by RT‐qPCR.THC and THC+CBD compromised initial learning of a repeated acquisition task. On the first day of daily treatment, THC alone induced emesis, but emesis was attenuated by the co‐administration of CBD. THC or THC+CBD suppressed locomotion and foraging; tolerance developed to the behaviorally suppressant effects during continuous daily exposure. Blood levels of THC and CBD remained relatively stable over 100 days of treatment. On the first day of treatment, levels of 11‐OH‐THC, an active THC metabolite, were lower in subjects that received THC+CBD than those that received THC alone; however this effect was not present at any other time point. Brain levels of THC were approximately 2 times higher than blood levels, and levels of THC were not affected significantly when CBD was co‐administered. Furthermore, daily THC or THC+CBD administered to adolescent monkeys for four months was associated with an altered mRNA transcriptome across multiple brain regions.CBD attenuates some but not all THC‐induced effects in nonhuman primates. Attenuation is unlikely to be related to disrupted THC metabolism or brain entry. Our findings warrant further research into the pharmacological and potential pathological consequences of THC and CBD after long term use, and a comparison of adolescent and adult responses to such cannabinoid exposure.Support or Funding InformationThis work was supported by NIDA grant DA042178.

Touchscreen technology in the study of cognition-related behavior.

There is a growing need for new translational animal models designed to capture complex behavioral phenotypes implicated in addiction and other neuropsychiatric conditions. For example, a complete understanding of the effects of commonly abused drugs, as well as candidate medications, requires assessments of their effects on learning, memory, attention, and other cognition-related behavior. Modern touch-sensitive technology provides an extremely flexible means to expose an experimental subject to a variety of complex behavioral tasks designed to assay dimensions of cognitive function before, during, and after drug administration. In addition to tailored variants of gold-standard cognitive assessments, touchscreen chambers offer the ability to develop novel tasks based upon the researcher's needs. This methods perspective presents (i) a brief review of previous touchscreen-based animal studies, (ii) a primer on the construction of a touch-sensitive experimental chamber, and (iii) data from a proof-of-concept study examining cross-species continuity in performance across a diverse assortment of animal subjects (rats, marmosets, squirrel monkeys, and rhesus macaques) using the repeated acquisition task - a modern variant of a traditional animal model of learning. Taken together, the procedures and data discussed in this review illustrate the point that contemporary touchscreen methodology can be tailored to desired experimental goals and adapted to provide formal similarity in cognition-related tasks across experimental species. Moreover, touchscreen methodology allows for the development of new translational models that emerge through laboratory and clinical discovery to capture important dimensions of complex behavior and cognitive function.

Touchscreen assays of learning, response inhibition, and motivation in the marmoset (Callithrix jacchus).

Recent developments in precision gene editing have led to the emergence of the marmoset as an experimental subject of considerable interest and translational value. A better understanding of behavioral phenotypes of the common marmoset will inform the extent to which forthcoming transgenic mutants are cognitively intact. Therefore, additional information regarding their learning, inhibitory control, and motivational abilities is needed. The present studies used touchscreen-based repeated acquisition and discrimination reversal tasks to examine basic dimensions of learning and response inhibition. Marmosets were trained daily to respond to one of the two simultaneously presented novel stimuli. Subjects learned to discriminate the two stimuli (acquisition) and, subsequently, with the contingencies switched (reversal). In addition, progressive ratio performance was used to measure the effort expended to obtain a highly palatable reinforcer varying in magnitude and, thereby, provide an index of relative motivational value. Results indicate that rates of both acquisition and reversal of novel discriminations increased across successive sessions, but that rate of reversal learning remained slower than acquisition learning, i.e., more trials were needed for mastery. A positive correlation was observed between progressive ratio break point and reinforcement magnitude. These results closely replicate previous findings with squirrel monkeys, thus providing evidence of similarity in learning processes across nonhuman primate species. Moreover, these data provide key information about the normative phenotype of wild-type marmosets using three relevant behavioral endpoints.

Chronic dietary chlorpyrifos causes long-term spatial memory impairment and thigmotaxic behavior

Little is known about the long-term effects of chronic exposure to low-level organophosphate (OP) pesticides, and the role of neurotransmitter systems, other than the cholinergic system, in mediating OP neurotoxicity. In this study, rats were administered 5mg/kg/day of chlorpyrifos (CPF) for 6 months commencing at 3-months-of-age. The animals were examined 7 months later (at 16-months-of-age) for spatial learning and memory in the Morris water maze (MWM) and locomotor activity. In addition, we assessed the chronic effects of CPF on glutamatergic and gamma-aminobutyric acid (GABAergic) function using pharmacological challenges with dizocilpine (MK801) and diazepam. Impaired performance related to altered search patterns, including thigmotaxis and long-term spatial memory was noted in the MWM in animals exposed to CPF, pointing to dietary CPF-induced behavioral disturbances, such as anxiety. Twenty-four hours after the 31st session of repeated acquisition task, 0.1mg/kg MK801, an N-methyl-d-aspartate (NMDA) antagonist was intraperitoneally (i.p.) injected for 4 consecutive days. Decreased latencies in the MWM in the control group were noted after two sessions with MK801 treatment. Once the MWM assessment was completed, animals were administered 0.1 or 0.2mg/kg of MK801 and 1 or 3mg/kg of diazepam i.p., and tested for locomotor activity. Both groups, the CPF dietary and control, displayed analogous performance in motor activity. In conclusion, our data point to a connection between the long-term spatial memory, thigmotaxic response and CPF long after the exposure ended.

Effects of self-administered methamphetamine on discrimination learning and reversal in nonhuman primates.

Frequent exposure to methamphetamine has been reported to adversely influence cognitive behavior and, in particular, inhibitory control processes. The present studies were conducted in squirrel monkeys to assess the effects of daily intravenous methamphetamine self-administration on touch screen-based repeated acquisition and discrimination reversal tasks thought to reflect behavioral dimensions of, respectively, learning and response inhibition. First, stable methamphetamine-maintained behavior was established in each subject (0.35-1.6 mg/kg/session), and subsequently, a second daily session of discrimination learning was conducted (20 h later). Subjects first learned to discriminate between two simultaneously presented stimuli (acquisition) and, subsequently, to re-learn the discrimination with the contingencies switched (reversal). The role of the interval between self-administration and touch screen sessions was evaluated, as well as the effects of abrupt methamphetamine discontinuation. Results indicate that daily methamphetamine self-administration markedly disrupted the development of discrimination learning, initially requiring nearly twice the number of trials to master discriminations. The magnitude of adverse effects in individual subjects correlated to the level of daily methamphetamine intake. Importantly, however, behavioral disruption of discrimination learning was surmounted following remedial training. Once criterion levels of discrimination performance were achieved, subsequent development of reversal performance was largely unaffected except when the interval between self-administration and touch screen session was short and, thus, likely a result of methamphetamine’s direct effects. Discontinuation of methamphetamine produced no disruption in acquisition or reversal. These results indicate that self-administered methamphetamine can markedly disrupt learning processes and highlight key differences in its effects on different aspects of discrimination learning.

Effects of an α5GABAA inverse agonist on MK-801-induced learning deficits in an incremental repeated acquisition task.

N-methyl-D-aspartate receptors (NMDARs) are essential for several kinds of synaptic plasticity and play a critical role in learning and memory. Deficits in NMDAR functioning may be partially responsible for the learning and memory deficits associated with aging and numerous diseases. Administration of MK-801, a noncompetitive NMDAR antagonist, is commonly used as a preclinical model of NMDAR dysfunction. The objective of this study was to assess the effects of α5GABAA receptor inhibition on learning deficits in the incremental repeated acquisition (IRA) task induced by acute MK-801 administration. The IRA task, commonly used to examine factors that affect learning, begins with a single response and increments to progressively longer chains throughout a single session as behavior meets preset criteria. MK-801 (0.03-0.5 mg/kg, intraperitoneally), administered 10 min pretesting, produced a significant dose-dependent decrease in measures of IRA performance at doses greater than or equal to 0.25 mg/kg. The MK-801-induced deficit was attenuated after treatment with an α5GABAA receptor inverse agonist, L-655,708 (1 mg/kg, intraperitoneally). The present study provides the focus for, and supports the feasibility of, further in-depth definitive studies examining α5GABAA receptor inhibition as a suitable candidate for the attenuation of NMDAR-related deficits.

Effects of self‐administered methamphetamine on learning‐to‐learn in the squirrel monkey (661.1)

Long‐term chronic exposure to methamphetamine (MA) has been related to profound neural impact including, but not limited to, damage to monoamine nerve terminals. In turn, neural changes following repeated MA administration may be related to adverse effects on learning and other cognitive performance. However, the characterization of such neurobehavioral deficits remains incomplete. The present study assessed the effects of ongoing MA self‐administration on touchscreen‐based discrimination learning in nonhuman primates. Prior to daily sessions of MA self‐administration (i.e., 20‐hr after the previous day’s session), monkeys engaged in a repeated acquisition task in which changes in the rate of discrimination learning for a stimulus pair were assessed over the successive presentation of novel stimulus pairs. Results thus far indicate that ongoing MA self‐administration produces markedly deleterious effects on the development of learning sets in discrimination learning. Importantly, the magnitude of adverse effects on learning appears to be correlated with the level of daily MA intake among individual subjects.Grant Funding Source: Supported by grant K01‐DA035974 from NIDA

Rescue of MK-801–induced learning deficits in an incremental repeated acquisition task using an α5 GABA(A) inverse agonist

Rescue of MK-801–induced learning deficits in an incremental repeated acquisition task using an α5 GABA(A) inverse agonist

Repeated acquisition and discrimination reversal in the squirrel monkey (Saimiri sciureus)

Repeated acquisition and discrimination reversal tasks are often used to examine behavioral relations of, respectively, learning and cognitive flexibility. Surprisingly, despite their frequent use in cognitive neuroscience and behavioral pharmacology, variables that control performance under these two tasks have not been widely studied. The present studies were conducted to directly investigate the controlling variables in nonhuman primates. Squirrel monkeys were trained with a touchscreen variant of the repeated acquisition task in which a novel pair of S(+)/S(-) stimuli was presented daily. Subjects learned to discriminate the two stimuli (acquisition) and, subsequently, with the contingencies switched (reversal). Results indicate that rates of both acquisition and reversal learning increased across successive sessions, but that rate of reversal learning remained slower than acquisition learning, i.e., more trials were needed for mastery. Subsequent experiments showed this difference between the rate of learning novel discriminations and reversal was reliable for at least 5 days between acquisition and reversal and notwithstanding the interpolation of additional discriminations. Experimental analysis of the S(+)/S(-) elements of the tasks revealed that the difference in the rate of learning could not be attributed to a relatively aversive quality of the S(-) or to a relatively appetitive quality of the S(+), but, rather, to contextual control by the S(+)/S(-) stimulus complex. Thus, if either element (S(+) or S(-)) of the stimulus complex was replaced by a novel stimulus, the rate of acquisition approximated that expected with a novel stimulus pair. These results improve our understanding of fundamental features of discrimination acquisition and reversal.

Pharmacological and behavioral profile of N-[(3R)-1-azabicyclo[2.2.2]oct-3-yl]-6-chinolincarboxamide (EVP-5141), a novel α7 nicotinic acetylcholine receptor agonist/serotonin 5-HT3 receptor antagonist

Agonists of α7 nicotinic acetylcholine receptors (nAChRs) may have therapeutic potential for the treatment of cognitive deficits. This study describes the in vitro pharmacology of the novel α7 nAChR agonist/serotonin 5-HT3 receptor (5-HT3R) antagonist N-[(3R)-1-azabicyclo[2.2.2]oct-3-yl]-6-chinolincarboxamide (EVP-5141) and its behavioral effects. EVP-5141 bound to α7 nAChRs in rat brain membranes (K i = 270nM) and to recombinant human serotonin 5-HT3Rs (K i = 880nM) but had low affinity for α4β2 nAChRs (K i > 100μM). EVP-5141 was a potent agonist at recombinant rat and human α7 nAChRs expressed in Xenopus oocytes. EVP-5141 acted as 5-HT3R antagonist but did not block α3β4, α4β2, and muscle nAChRs. Rats trained to discriminate nicotine from vehicle did not generalize to EVP-5141 (0.3-30mg kg(-1), p.o.), suggesting that the nicotine cue is not mediated by the α7 nAChR and that EVP-5141 may not share the abuse liability of nicotine. EVP-5141 (0.3-3mg kg(-1)) improved performance in the rat social recognition test. EVP-5141 (0.3mg kg(-1), p.o.) ameliorated scopolamine-induced retention deficits in the passive avoidance task in rats. EVP-5141 (1mg kg(-1), i.p.) improved spatial working memory of aged (26- to 32-month-old) rats in a water maze repeated acquisition task. In addition, EVP-5141 improved both object and social recognition memory in mice (0.3mg kg(-1), p.o.). EVP-5141 improved performance in several learning and memory tests in both rats and mice, supporting the hypothesis that α7 nAChR agonists may provide a novel therapeutic strategy for the treatment of cognitive deficits in Alzheimer's disease or schizophrenia.

A novel touch-sensitive apparatus for behavioral studies in unrestrained squirrel monkeys

Despite the increasing sophistication and affordability of touch-sensitive technology, its use in the behavioral sciences has been limited. The present paper describes the design and empirical validation of a novel touch-sensitive operant conditioning chamber for use with unrestrained squirrel monkeys. In addition, results from a variant of a commonly employed animal model of learning, the repeated acquisition task, demonstrate the effectiveness of this chamber in programming an assay of complex behavior. Finally, results from a study with Δ9-tetrahydrocannabinol, the active ingredient in marijuana, show that its effects in this novel touchscreen chamber were consistent with its dose-related effects on learning using more conventional approaches. Overall, these studies indicate the touchscreen apparatus provides effective means for programming complex behavioral tasks to assess the effects of pharmacological agents on cognitive function.

The use of an incremental repeated acquisition task to assess learning in children

The development of valid animal models of learning is especially important since learning is critical for nearly all aspects of human behavior and identifying appropriate surrogates provides additional opportunity to study various aspects of learning. Examining the factors that affect learning is often complicated by the need to administer the same task repeatedly across experimental conditions. Incremental repeated acquisition (IRA) tasks have been used extensively in animal research because they circumvent this problem by requiring a subject to learn different response chains repeatedly across sessions. The present study examined the association of age, sex of the participant, and IQ on the performance of an incremental repeated acquisition task in 837 children, aged 5-13 years. This task required children to learn to press four response levers in a specific sequence that was randomly chosen. Illumination of colored indicator lights signaled position in the required response chain. Initially, for the first link, only one of the four levers was correct: a response to it resulted in the delivery of a monetary reinforcer (5 cents). After mastery of the first link (i.e. three correct presses), the children were presented with a two-link response chain: a different lever had to be pressed before pressing the previously correct lever. After mastery of the two-link chain, the response chain length was once again increased, and so on until a response chain consisting of six links was completed or until the task timed out. Older children and children with higher IQs mastered longer response chain lengths and were more accurate in performance of this learning task than younger children. In addition, older children and children with higher IQs had higher effective response rates and lower ineffective response rates. No significant effects of the sex of the participant were demonstrated for any of the variables on this task, except overall response rate. The results indicate that this test is sensitive to developmental variables in children, with the degree of sensitivity of certain dependent variables being age-dependent. Characterization of performance of this task by humans facilitates comparisons with animal models employing the same task, thus enhancing its translational utility.

The Effects of Stimulant Medication on Performance of an Incremental Repeated Acquisition Task in Children with ADHD

The Effects of Stimulant Medication on Performance of an Incremental Repeated Acquisition Task in Children with ADHD

Effects of chronic oral acrylamide exposure on incremental repeated acquisition (learning) task performance in Fischer 344 rats

Acrylamide (ACR) is a relatively potent neurotoxicant. The ingestion of carbohydrate-containing foods cooked at high temperature exposes humans to low levels of ACR virtually daily. At relatively high levels of exposure (i.e., sub-chronic through chronic levels of exposure of ≥20mg/kg body weight/day), ACR has been shown in both rats and humans to produce a variety of effects on the nervous system. The possibility that chronic dietary exposure to ACR might produce brain toxicity which could impede the development of learning skills is a question of current concern. This research evaluated the effects of ACR on learning task performance in Fischer 344 rats exposed daily beginning prenatally and continuing throughout the lifespan. Dams were gavaged with ACR since implantation [gestation day (GD) 6] with 0, 0.1, 0.3, 1.0 or 5.0mg/kg/day through parturition. Gavaging continued at the same doses to pups through weaning on post-natal day (PND) 22 after which dosing continued via their drinking water. One male and one female per litter (8–9 per treatment group) were tested. Using an incremental repeated acquisition (IRA) task to assess learning ability, ACR-exposed Fischer 344 rats exhibited altered performance by 4months of age. From approximately 1–8months of age (through ~PND 240), over 52 testing sessions, a significant treatment effect was found on percent task completed (PTC), with Tukey's post-hoc test (P<0.05) indicating a significantly lower PTC for the 5.0mg/kg/day group compared to controls. While there was no treatment effect on accuracy (P=0.53), a significant decrease in response rate was seen at 5.0mg/kg/day, suggesting that the noted decrease in PTC was due to a decrease in rate of responding, not to an effect on accuracy of task solution. Previous findings in these same animals performing a progressive ratio task for the assessment of motivation, as well as a range of tests of motoric function, suggest that this decreased response rate could be due to subtle motoric effects, or possibly due to decreases in psychomotor speed, but is most likely due to motivational effects.

Interaction of nicotinic and histamine H 3 systems in the radial-arm maze repeated acquisition task

Nicotinic systems have been found in a variety of studies to play important roles in cognitive function. Nicotinic involvement in different aspects of cognitive function such as learning vs. memory may differ. We have found in rats that the spatial repeated acquisition task in the radial-arm maze is significantly improved by low doses of the nicotinic receptor antagonist mecamylamine, the atypical nicotinic receptor ligand lobeline, as well as the α7 nicotinic receptor agonist ARR-17779. Interestingly, nicotine in the same dose range that improves working memory in the win-shift radial maze task was not effective in improving repeated acquisition performance. Nicotinic systems interact with a variety of other neural systems. Differential involvement of these extended effects with learning vs. memory may help explain differential effects of nicotinic drugs with these cognitive functions. Histamine H 3 receptor antagonists have been shown by some studies to improve cognitive function, but others have not found this effect and some have found impairment. Nicotine stimulates the release of histamine. This effect may counter other cascading effects of nicotine in the performance of learning and memory tasks. A specific test of this hypothesis involves our study of nicotine (0.1–0.4 mg/kg) interactions with the histamine H 3 receptor antagonist thioperamide (2.5–10 mg/kg) on learning memory in the repeated acquisition test in the radial-arm maze. The highest dose of thioperamide tested caused a significant choice accuracy impairment, which was most evident during the later portions of the learning curve. The highest dose of nicotine did not change overall errors but did cause a significant impairment in learning over trials. The choice accuracy impairment induced by thioperamide was significantly attenuated by nicotine (0.4 mg/kg). The learning impairment caused by the highest dose of nicotine was significantly attenuated by thioperamide. Thioperamide also caused a slowing of response, an effect, which was attenuated by nicotine co-administration. The repeated acquisition test can help differentiate acute drug effects on learning. Nicotine and thioperamide effectively reversed each other's choice accuracy impairment even though each by itself impaired accuracy.

Noncholinergic Lesions of the Medial Septum Impair Sequential Learning of Different Spatial Locations

The medial septum and diagonal band of Broca (MSDB) are major afferents to the hippocampus and are important for learning, memory, and hippocampal theta rhythm. In the present study, we assessed the effect of cholinergic or noncholinergic MSDB lesions on the sequential learning of different goal locations in the same environment, a type of task that is proposed to require hippocampal theta rhythm. Rats were administered saline, 192-IgG saporin (SAP), or kainic acid (KA) into the MSDB and then behaviorally tested. On any day, a single arm of a radial maze was rewarded with food, but the location of this rewarded arm changed between days. As in previous studies, intraseptal SAP reduced the number of cholinergic neurons although sparing GABAergic septohippocampal neurons. KA had the reverse effect, reducing GABAergic septohippocampal neurons and sparing cholinergic neurons. KA, but not SAP, impaired performance on the repeated acquisition task. Saline and SAP rats showed rapid within-session learning, whereas KA rats were much slower to learn the goal location. Performance on a 30 min retention trial was also impaired, although this may be attributable to incomplete acquisition. These findings provide evidence that noncholinergic, but not cholinergic, MSDB neurons are important in helping the animal deal with high loads of memory interference, and provides partial support for the idea that hippocampal theta rhythm is involved.

Influence of seasonally adjusted exposure to cold and darkness on cognitive performance in circumpolar residents

The effects of seasonally adjusted 24-h exposure to cold and darkness on cognitive performance in urban circumpolar residents was assessed in 15 male subjects who spent three 24-h periods in a climatic chamber at 65 degrees latitude during the winter (January-March) and/or summer (August-September). Each subject was exposed to three different environmental conditions in random order: (1) 22 degrees C temperature and 500 lx lighting; (2) 10 degrees C temperature and 500 lx lighting; and (3) 10 degrees C temperature and 0.5-l lx lighting. Accuracy on an addition-subtraction task was significantly greater in the summer than in the winter (p= 0.038), while accuracy on a repeated acquisition task was significantly greater in the winter than in the summer (p < 0.001). Independent of season, exposure to cold and darkness was significantly associated with a decline in response time on five cognitive tests, an improvement in accuracy on three tests measuring complex cognitive tasks, and a decline in accuracy on two tests measuring simple tasks. Increased performance on complex tasks may result from increased arousal in response to the combination of cold temperatures and dim light characteristic of the winter in urban circumpolar settings.

Long-Term Neurotoxicity of Chlorpyrifos: Spatial Learning Impairment on Repeated Acquisition in a Water Maze

Organophosphate compounds are cholinesterase inhibitors widely used in agriculture, industry, household products, and even as chemical weapons. Their major mechanism of acute toxic action is the inhibition of acetylcholinesterase, which is responsible for the degradation of the neurotransmitter acetylcholine. An organophosphorus ester-induced chronic neurotoxicity (OPICN) syndrome has been proposed. The OPICN syndrome could result from both long-term exposure to subclinical doses of OPs and after acute poisoning. Development of animal models for the cognitive decline are required and could later help to elucidate the mechanisms involved in this long-term effect on the central nervous system. Previously, we have found performance decrements in a four-trial repeated acquisition spatial task in a water maze. The present study includes two experiments to extend the long-term behavioral effects observed. Rats were injected either once or twice with chlorpyrifos (CPF) and then tested months after in a two-trial repeated acquisition task in a water maze. Our results confirm and extend the long-term behavioral effects of subcutaneous administration of CPF. The two treatments used produced performance decrements that suggest functional central nervous system alterations.

Absence of long-term behavioral effects after sub-chronic administration of low doses of methamidophos in male and female rats

Putative long-term learning and memory effects of low-dose exposure to the cholinesterase inhibitor organophosphate methamidophos (Tamaron®) early in life were studied in two parallel studies in middle-aged rats. Methamidophos was administered via the drinking water to female and male Wistar rats using nominal concentrations of 0 (control), 0.5, 1.5 and 4.5 ppm active ingredient for 16 weeks. Animals were then maintained for a recovery period of about 14 months without treatment. They were tested in the standard and repeated acquisition version of the Morris water escape task in two series of tests starting 33 and 55 weeks after termination of the methamidophos treatment. Functional observations and motor activity measurements preceded each series of testing. Exposure to methamidophos was confirmed by measurement of brain cholinesterase (ChE-B) at the end of the 16 weeks of treatment in satellite animals. At 4.5 ppm a biologically relevant reduction in ChE-B activity was observed without clinical signs of intoxication (males: 66%, females: 64% of control activity). Mid- and low-dose exposure to methamidophos revealed ChE-B activity of 90% and 100% in males and 88% and 97% in females, respectively. General examinations of the animals during treatment revealed no clinical signs suggesting cholinergic stimulation. Functional observations and motor activity measurements exhibited no relevant differences between treatment groups and controls. Neither the performance in the standard Morris water escape task that predominantly measures spatial reference memory, nor in the repeated acquisition task in the Morris tank, which predominantly measures spatial working memory, was affected by treatment with methamidophos. A small number of statistically significant differences were noted in the mean performance level between treatment groups, or between treatment by sex groups in both versions of the Morris task. However, these findings appeared to be idiosyncratic for a particular experiment and were not supported by findings from the other. They were consequently not considered as reflecting a consistent effect of methamidophos on learning and memory. In conclusion, administration of low doses of methamidophos to female and male Wistar rats for 16 weeks during early adulthood did not impair spatial working and reference memory in the Morris water escape task 33 and 55 weeks after cessation of treatment.

Migratory sleeplessness in the white-crowned sparrow (Zonotrichia leucophrys gambelii).

Twice a year, normally diurnal songbirds engage in long-distance nocturnal migrations between their wintering and breeding grounds. If and how songbirds sleep during these periods of increased activity has remained a mystery. We used a combination of electrophysiological recording and neurobehavioral testing to characterize seasonal changes in sleep and cognition in captive white-crowned sparrows (Zonotrichia leucophrys gambelii) across nonmigratory and migratory seasons. Compared to sparrows in a nonmigratory state, migratory sparrows spent approximately two-thirds less time sleeping. Despite reducing sleep during migration, accuracy and responding on a repeated-acquisition task remained at a high level in sparrows in a migratory state. This resistance to sleep loss during the prolonged migratory season is in direct contrast to the decline in accuracy and responding observed following as little as one night of experimenter-induced sleep restriction in the same birds during the nonmigratory season. Our results suggest that despite being adversely affected by sleep loss during the nonmigratory season, songbirds exhibit an unprecedented capacity to reduce sleep during migration for long periods of time without associated deficits in cognitive function. Understanding the mechanisms that mediate migratory sleeplessness may provide insights into the etiology of changes in sleep and behavior in seasonal mood disorders, as well as into the functions of sleep itself.