Highlights • Zebrafish have advanced genetic and functional imaging toolkits to study sleep. • Zebrafish have conserved sleep genes and neurons. • Recent zebrafish studies have identified novel ...regulators of sleep and arousal. • Behavioral drug screens can identify links between disease and sleep disruption.
Sleep is a nearly universal feature of animal behaviour, yet many of the molecular, genetic, and neuronal substrates that orchestrate sleep/wake transitions lie undiscovered. Employing a viral ...insertion sleep screen in larval zebrafish, we identified a novel gene,
(
), whose loss results in behavioural hyperactivity and reduced sleep at night. The neuronally expressed
gene is conserved across vertebrates and encodes a small single-pass transmembrane protein that is structurally similar to the Na
,K
-ATPase regulator, FXYD1/Phospholemman. Disruption of either
or
, a Na
,K
-ATPase alpha-3 subunit associated with several heritable movement disorders in humans, led to decreased night-time sleep. Since
and
mutants have elevated intracellular Na
levels and non-additive effects on sleep amount at night, we propose that Dmist-dependent enhancement of Na
pump function modulates neuronal excitability to maintain normal sleep behaviour.
Novel invertebrate‐killing compounds are required in agriculture and medicine to overcome resistance to existing treatments. Because insecticides and anthelmintics are discovered in phenotypic ...screens, a crucial step in the discovery process is determining the mode of action of hits. Visible whole‐organism symptoms are combined with molecular and physiological data to determine mode of action. However, manual symptomology is laborious and requires symptoms that are strong enough to see by eye. Here, we use high‐throughput imaging and quantitative phenotyping to measure Caenorhabditis elegans behavioral responses to compounds and train a classifier that predicts mode of action with an accuracy of 88% for a set of ten common modes of action. We also classify compounds within each mode of action to discover substructure that is not captured in broad mode‐of‐action labels. High‐throughput imaging and automated phenotyping could therefore accelerate mode‐of‐action discovery in invertebrate‐targeting compound development and help to refine mode‐of‐action categories.
Synopsis
A combination of imaging and machine learning is used to predict compound mode of action using the unique behavioural responses of the roundworm C. elegans to different pesticides and anthelmintics.
Insecticides affect phenotypes in multiple behavioural dimensions.
Compounds with the same mode of action have similar effects on behaviour.
Combining classifiers by voting enables mode of action prediction.
The approach allows mode of action deconvolution within classes.
A combination of imaging and machine learning is used to predict compound mode of action using the unique behavioural responses of the roundworm C. elegans to different pesticides and anthelmintics.
Tracking small laboratory animals such as flies, fish, and worms is used for phenotyping in neuroscience, genetics, disease modelling, and drug discovery. An imaging system with sufficient throughput ...and spatiotemporal resolution would be capable of imaging a large number of animals, estimating their pose, and quantifying detailed behavioural differences at a scale where hundreds of treatments could be tested simultaneously. Here we report an array of six 12-megapixel cameras that record all the wells of a 96-well plate with sufficient resolution to estimate the pose of C. elegans worms and to extract high-dimensional phenotypic fingerprints. We use the system to study behavioural variability across wild isolates, the sensitisation of worms to repeated blue light stimulation, the phenotypes of worm disease models, and worms' behavioural responses to drug treatment. Because the system is compatible with standard multiwell plates, it makes computational ethological approaches accessible in existing high-throughput pipelines.
Localizing messenger RNAs at specific subcellular sites is a conserved mechanism for targeting the synthesis of cytoplasmic proteins to distinct subcellular domains, thereby generating the asymmetric ...protein distributions necessary for cellular and developmental polarity. However, the full range of transcripts that are asymmetrically distributed in specialized cell types, and the significance of their localization, especially in the nervous system, are not known. We used the EP-MS2 method, which combines EP transposon insertion with the MS2/MCP in vivo fluorescent labeling system, to screen for novel localized transcripts in polarized cells, focusing on the highly branched Drosophila class IV dendritic arborization neurons. Of a total of 541 lines screened, we identified 55 EP-MS2 insertions producing transcripts that were enriched in neuronal processes, particularly in dendrites. The 47 genes identified by these insertions encode molecularly diverse proteins, and are enriched for genes that function in neuronal development and physiology. RNAi-mediated knockdown confirmed roles for many of the candidate genes in dendrite morphogenesis. We propose that the transport of mRNAs encoded by these genes into the dendrites allows their expression to be regulated on a local scale during the dynamic developmental processes of dendrite outgrowth, branching, and/or remodeling.
Sleep is a nearly universal feature of animal behaviour, yet many of the molecular, genetic, and neuronal substrates that orchestrate sleep/wake transitions lie undiscovered. Employing a viral ...insertion sleep screen in larval zebrafish, we identified a novel gene,
dreammist
(
dmist
), whose loss results in behavioural hyperactivity and reduced sleep at night. The neuronally expressed
dmist
gene is conserved across vertebrates and encodes a small single-pass transmembrane protein that is structurally similar to the Na
+
,K
+
-ATPase regulator, FXYD1/Phospholemman. Disruption of either
fxyd1
or
atp1a3a
, a Na
+
,K
+
-ATPase alpha-3 subunit associated with several heritable movement disorders in humans, led to decreased night-time sleep. Since
atpa1a3a
and
dmist
mutants have elevated intracellular Na
+
levels and non-additive effects on sleep amount at night, we propose that Dmist-dependent enhancement of Na
+
pump function modulates neuronal excitability to maintain normal sleep behaviour.
Data collected for the eLife OpenAccess paper: Systematic creation and phenotyping of Mendelian disease models in C. elegans: towards large-scale drug repurposing. (doi: 10.7554/eLife.92491.1)
...Contains: extracted features, calculated stats, normalised z-scores and timerseries data of all the disease model mutants generated. In addition, there is a static .html file that allows for mousing over the clustermaps to easily view differences in strains compared to the N2 wild-type. Dataset also contains, metadata and feature summary/file name information of FDA-library drug screen and the confirmation screen of the hit from this (i.e., all data collected in published in the associated paper).
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