NemoTrainer: Automated Conditioning for Stimulus-Directed Navigation and Decision Making in Free-Swimming Zebrafish.

Current methods for associative conditioning in animals involve human intervention that is labor intensive, stressful to animals, and introduces experimenter bias in the data. Here, we describe a simple apparatus and a flexible, microcontroller-based conditioning paradigm that minimizes human intervention. Our methodology exploits directed movement towards a target that depends on spatial working memory, including processing of sensory inputs, motivational drive, and attentional mechanisms. Within a stimulus-driven conditioning paradigm designed to train zebrafish, we present a localized pulse of light via LEDs and/or sounds via an underwater transducer. A webcam placed below a glass tank records fish-swimming behavior. For classical conditioning, animals simply associate a sound or light with an unconditioned stimulus, such as a small food reward presented at a fixed location, and swim towards that location to obtain a few grains of food dispensed automatically via a sensor-triggered, stepper motor. During operant conditioning, a fish must first approach a proximity sensor at a remote location and then swim to the reward location. For both types of conditioning, a timing-gated interrupt activates stepper motors via custom software embedded within a microcontroller (Arduino). "Ardulink", a Java facility, implements Arduino-computer communication protocols. In this way, a Java-based user interface running on a host computer can provide full experimental control. Alternatively, a similar level of control is achieved via an Arduino script communicating with an event-driven application controller running on the host computer. Either approach can enable precise, multi-day scheduling of training, including timing, location, and intensity of stimulus parameters; and the feeder. Learning can be tracked by monitoring turning, location, response times, and directional swimming of individual fish. This facilitates the comparison of performance within and across a cohort of animals. Our scheduling and control software and apparatus ("NemoTrainer") can be used to study multiple aspects of species-specific behaviors as well as the effects on them of various interventions.

A randomized placebo-controlled pilot trial shows that intranasal vasopressin improves social deficits in children with autism.

The social impairments of autism spectrum disorder (ASD) have a major impact on quality of life, yet there are no medications that effectively treat these core social behavior deficits. Preclinical research suggests that arginine vasopressin (AVP), a neuropeptide involved in promoting mammalian social behaviors, may be a possible treatment for ASD. Using a double-blind, randomized, placebo-controlled, parallel study design, we tested the efficacy and tolerability of a 4-week intranasal AVP daily treatment in 30 children with ASD. AVP-treated participants aged 6 to 9.5 years received the maximum daily target dose of 24 International Units (IU); participants aged 9.6 to 12.9 years received the maximum daily target dose of 32 IU. Intranasal AVP treatment compared to placebo enhanced social abilities as assessed by change from baseline in this phase 2 trial's primary outcome measure, the Social Responsiveness Scale, 2nd Edition total score (SRS-2 T score; F 1,20 = 9.853; P = 0.0052; ηp 2 = 33.0%; Cohen's d = 1.40). AVP treatment also diminished anxiety symptoms and some repetitive behaviors. Most of these findings were more pronounced when we accounted for pretreatment AVP concentrations in blood. AVP was well tolerated with minimal side effects. No AVP-treated participants dropped out of the trial, and there were no differences in the rate of adverse events reported between treatment conditions. Last, no changes from baseline were observed in vital signs, electrocardiogram tracings, height and body weight, or clinical chemistry measurements after 4 weeks of AVP treatment. These preliminary findings suggest that AVP has potential for treating social impairments in children with ASD.

Intranasal oxytocin treatment for social deficits and biomarkers of response in children with autism.

Autism spectrum disorder (ASD) is characterized by core social deficits. Prognosis is poor, in part, because existing medications target only associated ASD features. Emerging evidence suggests that the neuropeptide oxytocin (OXT) may be a blood-based biomarker of social functioning and a possible treatment for ASD. However, prior OXT treatment trials have produced equivocal results, perhaps because of variability in patients' underlying neuropeptide biology, but this hypothesis has not been tested. Using a double-blind, randomized, placebo-controlled, parallel design, we tested the efficacy and tolerability of 4-wk intranasal OXT treatment (24 International Units, twice daily) in 32 children with ASD, aged 6-12 y. When pretreatment neuropeptide measures were included in the statistical model, OXT compared with placebo treatment significantly enhanced social abilities in children with ASD [as measured by the trial's primary outcome measure, the Social Responsiveness Scale (SRS)]. Importantly, pretreatment blood OXT concentrations also predicted treatment response, such that individuals with the lowest pretreatment OXT concentrations showed the greatest social improvement. OXT was well tolerated, and its effects were specific to social functioning, with no observed decrease in repetitive behaviors or anxiety. Finally, as with many trials, some placebo-treated participants showed improvement on the SRS. This enhanced social functioning was mirrored by a posttreatment increase in their blood OXT concentrations, suggesting that increased endogenous OXT secretion may underlie this improvement. These findings indicate that OXT treatment enhances social abilities in children with ASD and that individuals with pretreatment OXT signaling deficits may stand to benefit the most from OXT treatment.