Daniel T. Cerutti

Anxiolytic effects of nicotine in zebrafish

Anxiolytic effects of nicotine have been documented in studies with rodents and humans. Understanding the neural basis of nicotine-induced anxiolysis can help both with developing better aids for smoking cessation as well as with the potential development of novel nicotinic ligands for treating anxiety. Complementary non-mammalian models may be useful for determining the molecular bases of nicotine effects on neurobehavioral function. The current project examined whether a zebrafish model of anxiety would be sensitive to nicotine. When zebrafish are placed in a novel environment, they dive to the bottom of the tank. In the wild, diving could help to escape predation. We tested the anxiolytic effect of nicotine on the novelty-elicited diving response and subsequent habituation. Zebrafish placed in a novel tank spent the majority of time at the bottom third of the tank during the first minute of a 5-min session and then show a gradual decrease in time spent at the tank bottom. Nicotine treatment at 100 mg/l for 3 min by immersion before testing caused a significant decrease in diving throughout the session, while 50 mg/l was effective during the first minute when the greatest bottom dwelling was seen in controls. Nicotine effects were reversed by the nicotinic antagonist mecamylamine given together with nicotine, but not when administered shortly before the test session after prior nicotine dosing. This implies that the effect of nicotine on diving was due to net stimulation at nicotinic receptors, an effect that is blocked by mecamylamine; and that once invoked, this effect is no longer dependent on continuing activation of nicotinic receptors. The effect of nicotine on diving did not seem to be the result of a general disorientation of the fish. The 100 mg/ml nicotine dose was shown in our earlier study to significantly improve spatial-discrimination learning in zebrafish. Nicotine-induced anxiolytic effects can be modeled in the zebrafish. This preparation will help in the investigation of the molecular bases of this effect.

Zebrafish provide a sensitive model of persisting neurobehavioral effects of developmental chlorpyrifos exposure: Comparison with nicotine and pilocarpine effects and relationship to dopamine deficits

Chlorpyrifos (CPF) an organophosphate pesticide causes persisting behavioral dysfunction in rat models when exposure is during early development. In earlier work zebrafish were used as a complementary model to study mechanisms of CPF-induced neurotoxicity induced during early development. We found that developmental (first five days after fertilization) chlorpyrifos exposure significantly impaired learning in zebrafish. However, this testing was time and labor intensive. In the current study we tested the hypothesis that persisting effects of developmental chlorpyrifos could be detected with a brief automated assessment of startle response and that this behavioral index could be used to help determine the neurobehavioral mechanisms for persisting CPF effects. The swimming activity of adult zebrafish was assessed by a computerized video-tracking device after a sudden tap to the test arena. Ten consecutive trials (1/min) were run to determine startle response and its habituation. Additionally, habituation recovery trials were run at 8, 32 and 128 min after the end of the initial trial set. CPF-exposed fish showed a significantly (p<0.025) greater overall startle response during the 10-trial session compared to controls (group sizes: Control N=40, CPF N=24). During the initial recovery period (8 min) CPF-exposed fish showed a significantly (p<0.01) greater startle response compared to controls. To elucidate the contributions of nicotinic and muscarinic acetylcholine receptors to developmental CPF-mediated effects, the effects of developmental nicotine and pilocarpine exposure throughout the first five days after fertilization were determined. Developmental nicotine and pilocarpine exposure significantly increased startle response, though nicotine (group sizes: Control N=32, 15 mM N=12, 25 mM N=20) was much more potent than pilocarpine (group sizes: Control N=20, 100 microM N=16, 1000 microM N=12). Neither was as potent as CPF for developmental exposure increasing startle response in adulthood. Lastly, developmental CPF exposure decreased dopamine and serotonin levels and increased transmitter turnover in developing zebrafish larvae (N=4 batches of 50 embryos/treatment). Only the decline in dopamine concentrations persisted into adulthood (group sizes: Control N=14, CPF N=13). This study shows that a quick automated test of startle can detect persisting neurobehavioral impairments caused by developmental exposure to CPF. This may be helpful in screening for persisting neurobehavioral defects from a variety of toxicants.

Nicotine effects on learning in zebrafish: the role of dopaminergic systems

RationaleNicotine improves cognitive function in a number of animal models including rats, mice, monkeys, and recently, zebrafish. The zebrafish model allows higher throughput and ease in discovering mechanisms of cognitive improvement.Materials and methodsTo further characterize the neural bases of nicotine effects on learning in zebrafish, we determined changes in dopaminergic systems that accompany nicotine-enhanced learning.ResultsNicotine improved learning and increased brain levels of dihydroxyphenylacetic acid (DOPAC), the primary dopamine metabolite. There was a significant correlation between choice accuracy and DOPAC levels. The nicotinic antagonist mecamylamine blocked the nicotine-induced increase in DOPAC concentrations, in line with our previous finding that mecamylamine reversed nicotine-induced learning improvement.ConclusionsDopamine systems are related to learning in zebrafish; nicotine exposure increases both learning rates and DOPAC levels; and nicotinic antagonist administration blocks nicotine-induced rises in DOPAC concentrations. Rapid cognitive assessment of drugs with zebrafish could serve as a useful screening tool for the development of new therapeutics for cognitive dysfunction.

Assessing spatial vision - automated measurement of the contrast-sensitivity function in the hooded rat.

The contrast-sensitivity function (CSF) provides a concise and thorough description of an organisms spatial vision; it is widely used to describe vision in animals and humans, to track developmental changes in vision, and to compare vision among different species. Despite the predominance of rats in neuroscience research, their vision is not thoroughly studied due to the complexity of psychophysical measurement and a generally held notion that rat vision is poor. We therefore designed an economical and rapid method to assess the hooded rats CSF, using a computer monitor to display stimuli and an infrared touch screen to record responses. A six-alternative forced-choice task presented trials in which a sine-wave grating (S+), varying in spatial frequency and contrast, was displayed at different locations along with five gray stimuli (S-). Nose pokes to the S+ but not the S- produced water reinforcers. Contrasts were tested at each spatial frequency with a simple adaptive procedure until stimulus detection fell below chance. Psychometric functions were obtained by maximum-likelihood fitting of a logistic function to the raw data, obtaining the threshold as the functions point of inflection. As in previous studies with rats, CSFs showed an inverse-U shape with peak sensitivity at 0.12 cyc/deg and acuity just under 1 cyc/deg. The results indicate the present computer-controlled behavioral testing device is a precise and efficient instrument to assess spatial visual function in rats.

The Behavioral Economics of Choice and Interval Timing

The authors propose a simple behavioral economic model (BEM) describing how reinforcement and interval timing interact. The model assumes a Weber-law-compliant logarithmic representation of time. Associated with each represented time value are the payoffs that have been obtained for each possible response. At a given real time, the response with the highest payoff is emitted. The model accounts for a wide range of data from procedures such as simple bisection, metacognition in animals, economic effects in free-operant psychophysical procedures, and paradoxical choice in double-bisection procedures. Although it assumes logarithmic time representation, it can also account for data from the time-left procedure usually cited in support of linear time representation. It encounters some difficulties in complex free-operant choice procedures, such as concurrent mixed fixed-interval schedules as well as some of the data on double bisection, which may involve additional processes. Overall, BEM provides a theoretical framework for understanding how reinforcement and interval timing work together to determine choice between temporally differentiated reinforcers.

Time-sharing in pigeons: Independent effects of gap duration, position and discriminability from the timed signal.

Previous data suggest that in a peak-interval procedure with gaps, memory for the pre-gap interval varies with the discriminability of the gap from the to-be-timed signal. Here we extend this finding by manipulating the pre-gap and gap intervals as well as the visual contrast between the gap and the to-be-timed signal. The delay in response function after the gap was found to vary with the duration and position of the gap. However, for each gap duration and position, the delay in response increased with the gap-signal contrast: at 60% gap-signal contrast pigeons continued to accumulate time during the gap, at 80% gap-signal contrast pigeons stopped timing during the gap, and at 100% gap-signal contrast pigeons reset their timing after the gap. Data are accounted for by a time-sharing model assuming two concurrent processes during the gap--time accumulation and memory decay controlled by the salience of the gap--whose interplay results in a continuum of responses in the gap procedure.

Immediacy versus anticipated delay in the time-left experiment: a test of the cognitive hypothesis.

In the time-left experiment (J. Gibbon & R. M. Church, 1981), animals are said to compare an expectation of a fixed delay to food, for one choice, with a decreasing delay expectation for the other, mentally representing both upcoming time to food and the difference between current time and upcoming time (the cognitive hypothesis). The results of 2 experiments support a simpler view: that animals choose according to the immediacies of reinforcement for each response at a time signaled by available time markers (the temporal control hypothesis). It is not necessary to assume that animals can either represent or subtract representations of times to food to explain the results of the time-left experiment.

Timing in Choice Experiments

In Experiment 1, pigeons chose between variable- and fixed-interval schedules. The timer for 1 schedule was reset by a reinforcement on that schedule or on either schedule. In both cases, the pigeons timed reinforcement on each schedule from trial onset. The data further suggest that their behavior reflects 2 independent processes: 1 deciding when a response should be emitted and responsible for the timing of the overall activity, and the other determining what this response should be and responsible for the allocation of behavior between the 2 response keys. Results from Experiment 2, which studied choice between 2 fixed-interval schedules, support those 2 conclusions. These results have implications for the study of operant choice in general.

Effects of MDMA, methamphetamine and methylphenidate on repeated acquisition and performance in rats

Repeated-acquisition procedures that include performance controls for effects not specific to acquisition permit the assessment of drug effects on learning on a within-subject, within-session basis. Despite the advantages of this methodology, few studies have examined effects of psychomotor stimulants on repeated acquisition in rodents. The purpose of the present study was to investigate the effects of methylenedioxymethamphetamine (MDMA, 0.3-10mg/kg), methamphetamine (MA, 0.1-3mg/kg) and methylphenidate (MPD,1-17 mg/kg) using repeated-acquisition procedures with performance controls in rats using a touch-screen apparatus. Rats were presented a 2x3 array of stimuli using a computer touch-screen and nose-pokes to target locations within the array were reinforced. In the acquisition component, the correct location changed across sessions, whereas during the performance component, the correct location was constant across sessions. All three drugs reduced accuracy of responding to target locations in a dose-dependent fashion. None of the compounds enhanced learning at any dose. MPD and MA produced significant disruptions of acquisition accuracy only at doses that also disrupted performance, but the 3mg/kg dose of MDMA impaired acquisition of target responding without affecting performance. The selective impairment of acquisition found in the present study adds to the evidence of learning and memory disruption produced by acute MDMA administration and raise questions about the mechanisms for these actions.

Timescale invariance and Weber's law in choice

Pigeons were exposed to concurrent schedules for which reinforcement was alternately available at different times for each of two choices. In Experiment 1 (in which reinforcement times progressed arithmetically), overall, but not relative, response rate was timescale invariant. In Experiment 2 (in which reinforcement times progressed geometrically and were more spaced out), there was temporal control at all reinforcement times, but the amplitude of left-right response alternation decreased as time in the trial increased. These results indicate that the temporal regulation of both overall and relative response rates conforms to Webers law although relative rate is heavily influenced by processes other than timing. It also adds support to the idea that overall and relative response rate reflects the operation of two independent processes.