The mammalian-type thioredoxin reductase 1 confers a high-light tolerance to the green alga Chlamydomonas reinhardtii.

Reactive oxygen species (ROS) can both act as a poison causing cell death and important signaling molecules among various organisms. Photosynthetic organisms inevitably produce ROS, making the appropriate elimination of ROS an essential strategy for survival. Interestingly, the unicellular green alga Chlamydomonas reinhardtii expresses a mammalian form of thioredoxin reductase, TR1, which functions as a ROS scavenger in animal cells. To investigate the properties of TR1 in C. reinhardtii, we generated TR1 knockout strains using CRISPR/Cas9-based genome editing. We found a reduced tolerance to high-light and ROS stresses in the TR1 knockout strains compared to the parental strain. In addition, the regulation of phototactic orientation, known to be regulated by ROS, was affected in the knockout strains. These results suggest that TR1 contributes to a ROS-scavenging pathway in C. reinhardtii.

Enhanced polarotaxis can explain water-entry behaviour of mantids infected with nematomorph parasites.

A wide range of parasites manipulate the behaviours of their hosts in order to complete their life cycle1. Alteration of phototaxis is thought to be involved in host manipulation in many cases2,3. However, very little is known about what features of the light (intensity, spectrum, polarization) alter behaviour. Here we report that arboreal mantids (Hierodula patellifera) infected by nematomorph parasites (Chordodes sp.) are attracted to horizontally polarized light, which could induce the mantids to enter water, where the parasites can then emerge and reproduce. In a two-choice test, infected mantids were attracted to horizontally but not vertically polarized light. Uninfected mantids were not attracted to either. In a field experiment, 14 infected mantids entered a deep pool, where the water surface strongly reflected horizontally polarized light. By contrast, only two mantids entered a shallow pool, where the surface reflection had higher light intensity but weaker polarization. To our knowledge, this is the first study demonstrating that a manipulative parasite can take advantage of its hosts' ability to perceive polarized light stimuli to alter host behaviour. VIDEO ABSTRACT.

In search of the spectral composition of an effective light trap for the mushroom pest Lycoriella ingenua (Diptera: Sciaridae).

Certain fungus gnats, like Lycoriella ingenua are notorious pests in agriculture, especially in mushroom production. While larvae cause mainly direct crop damage, adults are vectors of several dangerous fungal pathogens. To promote the development of pesticide-free management methods, such as light trapping, we measured the spectral sensitivity of L. ingenua compound eyes with electroretinography and performed two different behavioural experiments to reveal the wavelength dependence of phototaxis in this species. The spectral sensitivity of the compound eyes is bimodal with peaks at 370 nm (UV) and 526 nm (green). Behavioural experiments showed that attraction to light as a function of wavelength depends on light intensity. In our first experiment, where the minimal photon flux (105-109 photons/cm2/s) needed for eliciting a phototactic response was determined wavelength by wavelength, phototaxis was strongest in the green spectral range (~526 nm). In the other behavioural experiment, where wavelength preference was tested under a higher but constant light intensity (~1013 photons/cm2/s), the highest attraction was elicited by UV wavelengths (398 nm). Our results suggest that both UV and green are important spectral regions for L. ingenua thus we recommend to use both UV (~370-398 nm) and green (~526 nm) for trapping these insects.

Complexity and plasticity in honey bee phototactic behaviour.

The ability to move towards or away from a light source, namely phototaxis, is essential for a number of species to find the right environmental niche and may have driven the appearance of simple visual systems. In this study we ask if the later evolution of more complex visual systems was accompanied by a sophistication of phototactic behaviour. The honey bee is an ideal model organism to tackle this question, as it has an elaborate visual system, demonstrates exquisite abilities for visual learning and performs phototaxis. Our data suggest that in this insect, phototaxis has wavelength specific properties and is a highly dynamical response including multiple decision steps. In addition, we show that previous experience with a light (through exposure or classical aversive conditioning) modulates the phototactic response. This plasticity is dependent on the wavelength used, with blue being more labile than green or ultraviolet. Wavelength, intensity and past experience are integrated into an overall valence for each light that determines phototactic behaviour in honey bees. Thus, our results support the idea that complex visual systems allow sophisticated phototaxis. Future studies could take advantage of these findings to better understand the neuronal circuits underlying this processing of the visual information.

stim2b Knockout Induces Hyperactivity and Susceptibility to Seizures in Zebrafish Larvae.

In neurons, stromal interaction molecule (STIM) proteins regulate store-operated Ca2+ entry (SOCE) and are involved in calcium signaling pathways. However, STIM activity in neurological diseases is unclear and should be clarified by studies that are performed in vivo rather than in cultured cells in vitro. The present study investigated the role of neuronal Stim2b protein in zebrafish. We generated stim2b knockout zebrafish, which were fertile and had a regular lifespan. Using various behavioral tests, we found that stim2b-/- zebrafish larvae were hyperactive compared with wild-type fish. The mutants exhibited increases in mobility and thigmotaxis and disruptions of phototaxis. They were also more sensitive to pentylenetetrazol and glutamate treatments. Using lightsheet microscopy, a higher average oscillation frequency and higher average amplitude of neuronal Ca2+ oscillations were observed in stim2b-/- larvae. RNA sequencing detected upregulation of the annexin 3a and gpr39 genes and downregulation of the rrm2, neuroguidin, and homer2 genes. The latter gene encodes a protein that is involved in several processes that are involved in Ca2+ homeostasis in neurons, including metabotropic glutamate receptors. We propose that Stim2b deficiency in neurons dysregulates SOCE and triggers changes in gene expression, thereby causing abnormal behavior, such as hyperactivity and susceptibility to seizures.

From behavior to circuit modeling of light-seeking navigation in zebrafish larvae.

Bridging brain-scale circuit dynamics and organism-scale behavior is a central challenge in neuroscience. It requires the concurrent development of minimal behavioral and neural circuit models that can quantitatively capture basic sensorimotor operations. Here, we focus on light-seeking navigation in zebrafish larvae. Using a virtual reality assay, we first characterize how motor and visual stimulation sequences govern the selection of discrete swim-bout events that subserve the fish navigation in the presence of a distant light source. These mechanisms are combined into a comprehensive Markov-chain model of navigation that quantitatively predicts the stationary distribution of the fish's body orientation under any given illumination profile. We then map this behavioral description onto a neuronal model of the ARTR, a small neural circuit involved in the orientation-selection of swim bouts. We demonstrate that this visually-biased decision-making circuit can capture the statistics of both spontaneous and contrast-driven navigation.

Carotenoids in the eyespot apparatus are required for triggering phototaxis in Euglena gracilis.

Carotenoids are the most universal and most widespread pigments in nature. They have played pivotal roles in the evolution of photosensing mechanisms in microbes and of vision in animals. Several groups of phytoflagellates developed a photoreceptive organelle called the eyespot apparatus (EA) consisting of two separable components: the eyespot, a cluster of carotenoid-rich globules that acts as a reflector device, and actual photoreceptors for photobehaviors. Unlike other algal eyespots, the eyespot of Euglenophyta lacks reflective properties and is generally considered to act as a shading device for the photoreceptor (paraflagellar body, PFB) for major photomovements. However, the function of the eyespot of Euglenophyta has not yet been fully proven. Here, we report that the blocking carotenoid biosynthesis in Euglena gracilis by suppressing the phytoene synthase gene (crtB) caused a defect in eyespot function resulting in a loss of phototaxis. Raman spectroscopy and transmission electron microscopy suggested that EgcrtB-suppressed cells formed eyespot globules but had a defect in the accumulation of carotenoids in those packets. Motion analysis revealed the loss of phototaxis in EgcrtB-suppressed cells: a defect in the initiation of turning movements immediately after a change in light direction, rather than a defect in the termination of cell turning at the appropriate position due to a loss of the shading effect on the PFB. This study revealed that carotenoids are essential for light perception by the EA for the initiation of phototactic movement by E. gracilis, suggesting one possible photosensory role of carotenoids in the EA for the phototaxis.

Light Spot-Based Assay for Analysis of Drosophila Larval Phototaxis.

The larvae of Drosophila melanogaster show obvious light-avoiding behavior during the foraging stage. Drosophila larval phototaxis can be used as a model to study animal avoidance behavior. This protocol introduces a light-spot assay to investigate larval phototactic behavior. The experimental set-up includes two main parts: a visual stimulation system that generates the light spot, and an infrared light-based imaging system that records the process of larval light avoidance. This assay allows tracking of the behavior of larva before entering, during encountering, and after leaving the light spot. Details of larval movement including deceleration, pause, head casting, and turning can be captured and analyzed using this method.

A machine vision system for zooplankton behavioural studies: a case study on the phototactic behaviour of sea lice (Lepeophtheirus salmonis) during sound and ultrasound stimuli.

Machine vision represents an accurate and easily verifiable method for observing live organisms and this technology is constantly evolving in terms of accessibility and cost. Motivated by the complexity of observing small-sized aquatic organisms in experimental systems, and the difficulties related to real-time observation, sampling and counting without interfering with the organisms, we here present a new method for observing behaviour and dispersion of non-sessile zooplankton organisms using a custom-made tank with an associated machine vision system. The system was used in an experiment where the aim was to assess the effect of sound and ultrasound on the phototactic behaviour of copepodite stages of the salmon louse (Lepeophtheirus salmonis). The experimental set-up is described, including a triangular test tank designed to create a sound pressure gradient, a mechanized camera movement system and a vision system with dedicated image processing software.

Noncanonical Photodynamics of the Orange/Green Cyanobacteriochrome Power Sensor NpF2164g7 from the PtxD Phototaxis Regulator of Nostoc punctiforme.

Forward and reverse primary (<10 ns) and secondary (>10 ns) photodynamics of cyanobacteriochrome (CBCR) NpF2164g7 were characterized by global analysis of ultrafast broadband transient absorption measurements. NpF2164g7 is the most C-terminal bilin-binding GAF domain in the Nostoc punctiforme phototaxis sensor PtxD (locus Npun_F2164). Although a member of the canonical red/green CBCR subfamily phylogenetically, NpF2164g7 exhibits an orange-absorbing 15ZPo dark-adapted state instead of the typical red-absorbing 15ZPr dark-adapted state characteristic of this subfamily. The green-absorbing 15EPg photoproduct of NpF2164g7 is unstable, allowing this CBCR domain to function as a power sensor. Photoexcitation of the 15ZPo state triggers inhomogeneous excited-state dynamics with three spectrally and temporally distinguishable pathways to generate the light-adapted 15EPg state in high yield (estimated at 25-30%). Although observed in other CBCR domains, the inhomogeneity in NpF2164g7 extends far into secondary relaxation dynamics (10 ns -1 ms) through to formation of 15EPg. In the reverse direction, the primary dynamics after photoexcitation of 15EPg are qualitatively similar to those of other red/green CBCRs, but secondary dynamics involve a "pre-equilibrium" step before regenerating 15ZPo. The anomalous photodynamics of NpF2164g7 may reflect an evolutionary adaptation of CBCR sensors that function as broadband light intensity sensors.

Sensorimotor computation underlying phototaxis in zebrafish.

Animals continuously gather sensory cues to move towards favourable environments. Efficient goal-directed navigation requires sensory perception and motor commands to be intertwined in a feedback loop, yet the neural substrate underlying this sensorimotor task in the vertebrate brain remains elusive. Here, we combine virtual-reality behavioural assays, volumetric calcium imaging, optogenetic stimulation and circuit modelling to reveal the neural mechanisms through which a zebrafish performs phototaxis, i.e. actively orients towards a light source. Key to this process is a self-oscillating hindbrain population (HBO) that acts as a pacemaker for ocular saccades and controls the orientation of successive swim-bouts. It further integrates visual stimuli in a state-dependent manner, i.e. its response to visual inputs varies with the motor context, a mechanism that manifests itself in the phase-locked entrainment of the HBO by periodic stimuli. A rate model is developed that reproduces our observations and demonstrates how this sensorimotor processing eventually biases the animal trajectory towards bright regions.Active locomotion requires closed-loop sensorimotor co ordination between perception and action. Here the authors show using behavioural, imaging and modelling approaches that gaze orientation during phototaxis behaviour in larval zebrafish is related to oscillatory dynamics of a neuronal population in the hindbrain.

Chlorophyll derivatives enhance invertebrate red-light and ultraviolet phototaxis.

Chlorophyll derivatives are known to enhance vision in vertebrates. They are thought to bind visual pigments (i.e., opsins apoproteins bound to retinal chromophores) directly within the retina. Consistent with previous findings in vertebrates, here we show that chlorin e6 - a chlorophyll derivative - enhances photophobicity in a flatworm (Dugesia japonica), specifically when exposed to UV radiation (λ = 405 nm) or red light (λ = 660 nm). This is the first report of chlorophyll derivatives acting as modulators of invertebrate phototaxis, and in general the first account demonstrating that they can artificially alter animal response to light at a behavioral level. Our findings show that the interaction between chlorophyll derivatives and opsins virtually concerns the vast majority of bilaterian animals, and also occurs in visual systems based on rhabdomeric (rather than ciliary) opsins.

Influence of static magnetic fields in phototaxis and osmotic stress in Gymnodinium catenatum (Dinophyceae).

Phototaxis response of the toxic microalgae Gymnodinium catenatum was studied in vitro. The percentage of cells remaining at mid-depth 20 min after stirring increased with solar radio, X-ray and solar flares output. It also increased with geomagnetic activity and temperature, and was dependent on culture time. Increase in the local static magnetic field with a permanent magnet did not influence the positive phototaxis response. However, survival and growth to a provoked hypo-osmotic shock in an altered static magnetic field was dependent on culture time and geomagnetic activity at a threshold below 22 nT. The results from phototaxis and hypo-osmotic shock experiments were in line with the previous hypothesis for the existence of two separate deleterious mechanisms conditioning the natural blooms of G. catenatum: one that is dependent on solar radiation and the other that is related to geomagnetic activity. Variations in electromagnetic fields caused by tectonic activity were also capable of influencing G. catenatum phototaxis and growth response in vitro.

Cyanobacteria in motion.

Cyanobacteria are able to move directly towards or away from a light source, a process called phototaxis. Recent studies have revealed that the spherical unicellular cyanobacterium Synechocystis sp. PCC 6803 exhibits a cell polarity in response to unidirectional illumination and that micro-optic properties of cyanobacterial cells are the basis of their directional light sensing. Further functional and physiological studies highlight a very complex control of cyanobacterial phototaxis by sensory proteins, histidine kinases and response regulators. Notably, PATAN domain response regulators appear to participate in directional control of phototaxis in the cyanobacterium Synechocystis sp. PCC 6803. In this review we explain the problem of directional light sensing at the small scale of bacteria and discuss our current understanding of signal transduction in cyanobacterial phototaxis.

Protocols to Study Behavior in Drosophila.

Drosophila melanogaster is an incredibly versatile organism capable of both innate and higher-order behaviors. These behaviors offer not only a way to assay whether or not the animal is physiologically compromised (e.g., feeding, locomotion), but also serve to assess changes in centrally mediated functions. Here we describe several high throughput, reproducible, yet inexpensive and facile behavioral assays for both larval and adult Drosophila. The larval assays all employ an agar substrate in a petri dish; the adult assays are grouped into "vial-based" and "arena-based" paradigms. While these protocols are largely designed to assess individual animals, they are sufficiently rapid that ample numbers can be tested to determine behavioral significance. Importantly, this also allows for one to control for reproductive status, age, and sex, since these factors all have a significant impact on adult behaviors. In general, it is best to designate a dedicated area for any assay, so that lighting conditions are consistent, and all animals should be tested at roughly the same time each day to minimize circadian fluctuations. Temperature and humidity should also be maintained at a constant level to minimize variability in the assays.

Decision-making ability of Physarum polycephalum enhanced by its coordinated spatiotemporal oscillatory dynamics.

An amoeboid unicellular organism, a plasmodium of the true slime mold Physarum polycephalum, exhibits complex spatiotemporal oscillatory dynamics and sophisticated information processing capabilities while deforming its amorphous body. We previously devised an 'amoeba-based computer (ABC),' that implemented optical feedback control to lead this amoeboid organism to search for a solution to the traveling salesman problem (TSP). In the ABC, the shortest TSP route (the optimal solution) is represented by the shape of the organism in which the body area (nutrient absorption) is maximized while the risk of being exposed to aversive light stimuli is minimized. The shortness of the TSP route found by ABC, therefore, serves as a quantitative measure of the optimality of the decision made by the organism. However, it remains unclear how the decision-making ability of the organism originates from the oscillatory dynamics of the organism. We investigated the number of coexisting traveling waves in the spatiotemporal patterns of the oscillatory dynamics of the organism. We show that a shorter TSP route can be found when the organism exhibits a lower number of traveling waves. The results imply that the oscillatory dynamics are highly coordinated throughout the global body. Based on the results, we discuss the fact that the decision-making ability of the organism can be enhanced not by uncorrelated random fluctuations, but by its highly coordinated oscillatory dynamics.