Behavioural phenotyping of C. elegans on UV-killed E. coli mutants v1

Protocol for screening candidate behaviour-modifying E. coli BW25113 single-gene deletion mutants from the 'Keio Collection', to investigate their effects on Caenorhabditis elegans behaviour when killed by ultraviolet (UV) light

Quantitative behavioural phenotyping to investigate anaesthesia induced neurobehavioural impairment

Anaesthesia exposure to the developing nervous system causes neuroapoptosis and behavioural impairment in vertebrate models. Mechanistic understanding is limited, and target-based approaches are challenging. High-throughput methods may be an important parallel approach to drug-discovery and mechanistic research. The nematode worm Caenorhabditis elegans is an ideal candidate model. A rich subset of its behaviour can be studied, and hundreds of behavioural features can be quantified, then aggregated to yield a ‘signature’. Perturbation of this behavioural signature may provide a tool that can be used to quantify the effects of anaesthetic regimes, and act as an outcome marker for drug screening and molecular target research. Larval C. elegans were exposed to: isoflurane, ketamine, morphine, dexmedetomidine, and lithium (and combinations). Behaviour was recorded, and videos analysed with automated algorithms to extract behavioural features. Anaesthetic exposure during early development leads to persisting behavioural variation (in total, 125 features across exposure combinations). Higher concentrations, and combinations of isoflurane with ketamine, lead to persistent change in a greater number of features. Morphine and dexmedetomidine do not appear to lead to behavioural impairment. Lithium rescues the neurotoxic phenotype produced by isoflurane. Findings correlate well with vertebrate research: impairment is dependent on agent, is concentration-specific, is more likely with combination therapies, and can potentially be rescued by lithium. These results suggest that C. elegans may be an appropriate model with which to pursue phenotypic screens for drugs that mitigate the neurobehavioural impairment. Some possibilities are suggested for how high-throughput platforms might be organised in service of this field.

A non-invasive radar system for automated behavioural tracking: application to sheep

Automated quantification of the behaviour of freely moving animals is increasingly needed in ethology, ecology, genetics and evolution. State-of-the-art approaches often require tags to identify animals, high computational power for data collection and processing, and are sensitive to environmental conditions, which limits their large-scale utilisation. Here we introduce a new automated tracking system based on millimetre-wave radars for real time robust and high precision monitoring of untagged animals. To validate our system, we tracked 64 sheep in a standard indoor behavioural test used for genetic selection. First, we show that the proposed radar application is faster and more accurate than conventional video and infrared tracking systems. Next, we illustrate how new behavioural estimators can be derived from the radar data to assess personality traits in sheep for behavioural phenotyping. Finally, we demonstrate that radars can be used for movement tracking at larger spatial scales, in the field, by adjusting operating frequency and radiated electromagnetic power. Millimetre-wave radars thus hold considerable promises for high-throughput recording of the behaviour of animals with various sizes and locomotor modes, in different types of environments.

Development of Open-Field Behaviour in the Medaka, Oryzias latipes

The use of juvenile and larval fish models has been growing in importance for several fields. Accordingly, the evaluation of behavioural tests that can be applied to larvae and juveniles is becoming increasingly important. We tested medaka at four different ages (1, 10, 30, and 120 dph) in the open field test, one of the most commonly used behavioural assays, to investigate its suitability for larvae and juveniles of this species. We also explored ontogenetic variation in behaviour during this test. On average, adult 120-day-old medaka showed higher locomotor activity in terms of distance moved compared with younger fish. Our analysis suggests that this effect was derived from both quantitative changes in locomotion related to the ontogenetic increase in fish size as well as qualitative changes in two aspects of locomotor behaviour. Specifically, time spent moving was similar between 1- and 10-day-old medaka, but progressively increased with development. In addition, we revealed that adult medaka showed constant levels of activity, whereas younger medaka progressively reduced their activity over the course of the entire experiment. The thigmotaxis behaviour typically used to assess anxiety in the open field test emerged at 120 days post-hatching, even though a difference in the temporal pattern of spatial preference emerged earlier, between 10 and 30 days post-hatching. In conclusion, some measures of the open field test such as total distance moved allow behavioural phenotyping in the medaka of all ages, although with some degree of quantitative and qualitative developmental variation. In contrast, immature medaka appear not to exhibit thigmotactic behaviour.

Quantitative Behavioural Phenotyping to Investigate Anaesthesia-Induced Neurobehavioural Impairment

BackgroundVertebrate animal experiments have suggested that anaesthesia exposure to the developing nervous system causes neuroapoptosis and behavioural impairment. Mechanistic understanding is limited, and target-based approaches are challenging. High-throughput methods may be an important parallel approach to drug-discovery and mechanistic research. The nematode worm Caenorhabditis elegans is an ideal candidate as a model for this. A rich subset of its behaviour can be studied, and hundreds of morphological and behavioural features can be quantified, then aggregated to yield a ‘signature’. Perturbation of this behavioural signature may provide a tool that can be used to quantify the effects of anaesthetic regimes, and act as an outcome marker for drug screening and molecular target research.MethodsLarval C. elegans were exposed to: isoflurane, ketamine, morphine, dexmedetomidine, and lithium (and combinations). Behaviour was recorded, and videos analysed with automated algorithms to extract behavioural and morphological features.ResultsAnaesthetic exposure during early development leads to persisting morphological and behavioural variation (in total, 125 features/exposure combinations). Higher concentrations, and combinations of isoflurane with ketamine, lead to more feature ‘hits’. Morphine and dexmedetomidine do not appear to lead to behavioural impairment. Lithium rescues the neurotoxic phenotype produced by isoflurane.ConclusionsFindings correlate well with vertebrate research: impairment is dependent on agent, is concentration-specific, is more likely with combination therapies, and can potentially be rescued by lithium. These results suggest that C. elegans may be an appropriate model with which to pursue phenotypic screens for drugs that mitigate the neurobehavioural impairment. Some possibilities are suggested for how high-throughput platforms might be organised in service of this field.

Specific profile of ultrasonic communication in a mouse model of neurodevelopmental disorders

Mice emit ultrasonic vocalizations (USVs) in different social conditions: pups maternal separation, juveniles play, adults mating and social investigation. The USVs measurement has become an important instrument for behavioural phenotyping in neurodevelopmental disorders (NDDs). Recently, we have demonstrated that the deletion of the NFκB1 gene, which encodes the p50 NF-κB subunit, causes NDDs phenotype in mice. In this study, we investigated the ultrasonic communication and the effects of an early social enrichment in mice lacking the NF-κB p50 subunit (p50 KO). In particular, USVs of wild-type (WT), p50 KO and KO exposed to early social enrichment (KO enriched) were recorded using an ultrasound sensitive microphone and analysed by Avisoft software. USVs analysis showed that p50 KO pups emit more and longer vocalizations compared to WT pups. On the contrary, in adulthood, p50 KO mice emit less USVs than WT mice. We also found significant qualitative differences in p50 KO mice USVs compared to WT mice; the changes specifically involved two USVs categories. Early social enrichment had no effect on USVs number, duration and type in p50 KO mice. Together, these data revealed social communication alterations in a mouse model of NDDs; these deficits were not recovered by early social enrichment, strengthening the fact that genetic background prevails on environmental enrichment.

A Bayesian account of generalist and specialist formation under the Active Inference framework

This paper offers a formal account of policy learning, or habitual behavioural optimisation, under the framework of Active Inference. In this setting, habit formation becomes an autodidactic, experience-dependent process, based upon what the agent sees itself doing. We focus on the effect of environmental volatility on habit formation by simulating artificial agents operating in a partially observable Markov decision process. Specifically, we used a ‘two-step’ maze paradigm, in which the agent has to decide whether to go left or right to secure a reward. We observe that in volatile environments with numerous reward locations, the agents learn to adopt a generalist strategy, never forming a strong habitual behaviour for any preferred maze direction. Conversely, in conservative or static environments, agents adopt a specialist strategy; forming strong preferences for policies that result in approach to a small number of previously-observed reward locations. The pros and cons of the two strategies are tested and discussed. In general, specialization offers greater benefits, but only when contingencies are conserved over time. We consider the implications of this formal (Active Inference) account of policy learning for understanding the relationship between specialisation and habit formation.Author SummaryActive inference is a theoretical framework that formalizes the behaviour of any organism in terms of a single imperative – to minimize surprise. Starting from this principle, we can construct simulations of simple “agents” (artificial organisms) that show the ability to infer causal relationships and learn. Here, we expand upon currently-existing implementations of Active Inference by enabling synthetic agents to optimise the space of behavioural policies that they can pursue. Our results show that by adapting the probabilities of certain action sequences (which may correspond biologically to the phenomenon of synaptic plasticity), and by rejecting improbable sequences (synaptic pruning), the agents can begin to form habits. Furthermore, we have shown our agent’s habit formation to be environment-dependent. Some agents become specialised to a constant environment, while other adopt a more general strategy, each with sensible pros and cons. This work has potential applications in computational psychiatry, including in behavioural phenotyping to better understand disorders.