Phenotyping single-cell motility in microfluidic confinement

At all scales, the movement patterns of organisms serve as dynamic read-outs of their behaviour and physiology. We devised a novel droplet microfluidics assay to encapsulate single algal microswimmers inside closed arenas, and comprehensively studied their roaming behaviour subject to a large number of environmental stimuli. We compared two model species, Chlamydomonas reinhardtii (freshwater alga, 2 cilia), and Pyramimonas octopus (marine alga, 8 cilia), and detailed their highly-stereotyped behaviours and the emergence of a trio of macroscopic swimming states (smooth-forward, quiescent, tumbling or excitable backward). Harnessing ultralong timeseries statistics, we reconstructed the species-dependent reaction network that underlies the choice of locomotor behaviour in these aneural organisms, and discovered the presence of macroscopic non-equilibrium probability fluxes in these active systems. We also revealed for the first time how microswimmer motility changes instantaneously when a chemical is added to their microhabitat, by inducing deterministic fusion between paired droplets - one containing a trapped cell, and the other, a pharmacological agent that perturbs cellular excitability. By coupling single-cell entrapment with unprecedented tracking resolution, speed and duration, our approach offers unique and potent opportunities for diagnostics, drug-screening, and for querying the genetic basis of micro-organismal behaviour.

The effects of femoral metaphyseal morphology on growth plate biomechanics in juvenile chimpanzees and humans

The distal femoral metaphyseal surface presents dramatically different morphologies in juvenile extant hominoids—humans have relatively flat metaphyseal surfaces when compared with the more complex metaphyseal surfaces of apes. It has long been speculated that these different morphologies reflect different biomechanical demands placed on the growth plate during locomotor behaviour, with the more complex metaphyseal surfaces of apes acting to protect the growth plate during flexed-knee behaviours like squatting and climbing. To test this hypothesis, we built subject-specific parametric finite-element models from the surface scans of the femora of five Pan and six Homo juveniles. We then simulated the loading conditions of either a straight-leg or flexed-knee gait and measured the resulting stresses at the growth plate. When subjected to the simulated flexed-knee loading conditions, both the maximum and mean von Mises stresses were significantly lower in the Pan models than in the Homo models. Further, during these loading conditions, von Mises stresses were strongly negatively correlated with ariaDNE, a measure of complexity of the metaphyseal surface. These results indicate that metaphyseal surface morphology has a robust effect on growth plate mechanics.

Quantitative assessment of tarsal morphology illuminates locomotor behaviour in Palaeocene mammals following the end-Cretaceous mass extinction

Mammals exhibit vast ecological diversity, including a panoply of locomotor behaviours. The foundations of this diversity were established in the Mesozoic, but it was only after the end-Cretaceous mass extinction that mammals began to increase in body size, diversify into many new species and establish the extant orders. Little is known about the palaeobiology of the mammals that diversified immediately after the extinction during the Palaeocene, which are often perceived as ‘archaic’ precursors to extant orders. Here, we investigate the locomotor ecology of Palaeocene mammals using multivariate and disparity analyses. We show that tarsal measurements can be used to infer locomotor mode in extant mammals, and then demonstrate that Palaeocene mammals occupy distinctive regions of tarsal morphospace relative to Cretaceous and extant therian mammals, that is distinguished by their morphological robustness. We find that many Palaeocene species exhibit tarsal morphologies most comparable with morphologies of extant ground-dwelling mammals. Disparity analyses indicate that Palaeocene mammals attained similar morphospace diversity to the extant sample. Our results show that mammals underwent a post-extinction adaptive radiation in tarsal morphology relating to locomotor behaviour by combining a basic eutherian bauplan with anatomical specializations to attain considerable ecomorphological diversity.

Alarm Pheromone Responses Depend on Genotype, but Not on the Presence of Facultative Endosymbionts in the Pea Aphid Acyrthosiphon pisum

Aphids use an alarm pheromone, E-β farnesene (EBF), to warn conspecifics of potential danger. The antennal sensitivity and behavioural escape responses to EBF can be influenced by different factors. In the pea aphid, Acyrthosiphon pisum, different biotypes are adapted to different legume species, and within each biotype, different genotypes exist, which can carry or not Hamiltonella defensa, a bacterial symbiont that can confer protection against natural enemies. We investigate here the influence of the aphid genotype and symbiotic status on the escape behaviour using a four-way olfactometer and antennal sensitivity for EBF using electroantennograms (EAGs). Whereas the investigated three genotypes from two biotypes showed significantly different escape and locomotor behaviours in the presence of certain EBF doses, the infection with H. defensa did not significantly modify the escape behaviour and only marginally influenced the locomotor behaviour at high doses of EBF. Dose-response curves of EAG amplitudes after stimulation with EBF differed significantly between aphid genotypes in correlation with behavioural differences, whereas antennal sensitivity to EBF did not change significantly as a function of the symbiotic status. The protective symbiont H. defensa does thus not modify the olfactory sensitivity to the alarm pheromone. How EBF sensitivity is modified between genotypes or biotypes remains to be investigated.

The Effectiveness of Scutellaria baicalensis on Migraine: Implications from Clinical Use and Experimental Proof

Background. Scutellaria baicalensis (SB), a traditional Chinese medicine, is commonly used for the treatment of inflammatory and painful conditions. The purpose of the present study was to examine the effects of SB on migraine. Materials and Methods. We examined the clinical applications of SB based on the data obtained from Taiwan’s National Health Insurance Research Database and confirmed that it was frequently used in Taiwan for the treatment of headaches. An experimental migraine model was established in rats by an intraperitoneal injection of nitroglycerin (NTG, 10 mg/kg). Pretreatment with SB was given orally 30 min before NTG administration. The rats were subjected to migraine-related behaviour tests that were video-recorded and analysed using EthoVision XT 12.0 software. Results. The frequency of exploratory and locomotor behaviour was comparatively lower in the NTG group than that in the control group, while the frequency of resting and grooming behaviour increased. These phenomena were ameliorated by pretreatment with 1.0 g/kg SB. The total time spent on the smooth surface was longer in the NTG group than that in the control group, but the time was shortened by pretreatment with 1.0 g/kg SB. Conclusions. Pretreatment with 1.0 g/kg SB relieved migraine-related behaviours in the experimental NTG-induced migraine model. The outcome therefore demonstrated that pretreatment with 1.0 g/kg SB is beneficial for migraine treatment.

Tau-mediated axonal degeneration is prevented by activation of the WldS pathway

Tauopathy is characterised by neuronal dysfunction and degeneration occurring as a result of changes to the microtubule associated protein tau. The neuronal changes evident in Tauopathy bear striking morphological resemblance to those reported in models of Wallerian degeneration. The mechanisms underpinning Wallerian degeneration are not fully understood although it can be delayed by the expression of the slow Wallerian degeneration (WldS) protein, which has also been demonstrated to delay axonal degeneration in some models of neurodegenerative disease. Given the morphological similarities between tauopathy and Wallerian degeneration, this study investigated whether tau-mediated phenotypes can be modulated by expression of WldS. In a Drosophila model of tauopathy in which expression of human Tau protein (hTau0N3R) leads to progressive age-dependent phenotypes, activation of the pathway downstream of WldS completely suppressed tau-mediated degeneration. This protective effect was evident even if the pathway downstream of WldS was activated several weeks after hTau-mediated degeneration had become established. In contrast, WldS expression without activation of the downstream protective pathway did not rescue tau-mediated degeneration in adults or improve tau-mediated neuronal dysfunction including deficits in axonal transport, synaptic alterations and locomotor behaviour in hTau0N3R –expressing larvae. This collectively implies that the pathway mediating the protective effect of WldS intersects with the mechanism(s) of degeneration initiated by hTau and can effectively halt tau-mediated degeneration at both early and late stages. Understanding the mechanisms underpinning this protection could identify much-needed disease-modifying targets for tauopathies.

Comparative Gross Anatomy of the Forelimb Arteries of the Japanese Monkey (Macaca fuscata) and a Comparative Pattern of Forelimb Arterial Distribution in Primates

Macaca fuscata displays characteristic behaviours, such as stone handling, locomotor behaviour, gait position, and intermittent bipedalism. Differences in characteristic behaviours among primate species/genera could be explained by anatomical details of the body. However, the anatomical details have not been well studied in Macaca fuscata. Arterial models could be one of the anatomical bases for the phylogenetic and functional differences among species, since the arterial supply could be associated with the muscular performance, especially locomotor behaviour. In this study, five thoracic limbs of Macaca fuscata adults were dissected to analyse the vessels. Patterns of arterial distribution in the thoracic limbs of Macaca fuscata were compared with those in other primates. The results indicated that the arterial distribution in the Japanese monkeys was more similar to those in Macaca mulatta and Papio anubis, which is consistent with phylogenetic similarities. However, compared with Papio anubis and other macaques, there were anatomical differences in several points, including (1) the origin of the common, anterior, posterior circumflex, and profunda brachii, and (2) the origins of the collateralis ulnaris artery. The comparative anatomy of the arteries in the forelimb of Macaca fuscata, along with the anatomical studies in other primates, indicated characteristic patterns of brachial artery division and the number of the palmar arches in primates, which is consistent with the phylogenetic division among New World primates, Old World primates, and apes.

Opposing forms of adaptation in mouse visual cortex are controlled by distinct inhibitory microcircuits and gated by locomotion

SummaryCortical processing of sensory signals adjusts to changes in both the external world and the internal state of the animal. We investigated the neural circuitry by which these processes interact in the primary visual cortex of mice. An increase in contrast caused as many pyramidal cells (PCs) to sensitize as depress, reflecting the dynamics of adaptation in different types of interneuron (PV, SST and VIP). Optogenetic manipulations demonstrate that the net effect within PCs reflects the balance of PV inputs, driving depression, and a subset of SST interneurons, driving sensitization. Locomotor behaviour increased the gain of PC responses by disinhibition through both the VIP->SST and SST->PV pathways, thereby maintaining the balance between opposing forms of plasticity. These experiments reveal how inhibitory microcircuits interact to purpose different subsets of PCs for the signalling of increases or decreases in contrast while also allowing for behavioural control of gain across the population.