Current Methods to Investigate Nociception and Pain in Zebrafish

Pain is an unpleasant, negative emotion and its debilitating effects are complex to manage. Mammalian models have long dominated research on nociception and pain, but there is increasing evidence for comparable processes in fish. The need to improve existing pain models for drug research and the obligation for 3R refinement of fish procedures facilitated the development of numerous new assays of nociception and pain in fish. The zebrafish is already a well-established animal model in many other research areas like toxicity testing, as model for diseases or regeneration and has great potential in pain research, too. Methods of electrophysiology, molecular biology, analysis of reflexive or non-reflexive behavior and fluorescent imaging are routinely applied but it is the combination of these tools what makes the zebrafish model so powerful. Simultaneously, observing complex behavior in free-swimming larvae, as well as their neuronal activity at the cellular level, opens new avenues for pain research. This review aims to supply a toolbox for researchers by summarizing current methods to study nociception and pain in zebrafish. We identify treatments with the best algogenic potential, be it chemical, thermal or electric stimuli and discuss options of analgesia to counter effects of nociception and pain by opioids, non-steroidal anti-inflammatory drugs (NSAIDs) or local anesthetics. In addition, we critically evaluate these practices, identify gaps of knowledge and outline potential future developments.

Equilibria in the Stackelberg Oligopoly Reflexive Games with Different Marginal Costs of Agents

This paper considers the problem of searching for information equilibria in an oligopoly market in the case of Stackelberg leaders. A framework considers the reflexive behavior of three agents, and linear agent’s cost functions with different coefficients (i.e., marginal and fixed costs) are considered. The results of the study are as follows. First, models of the reflexive games for a triopoly that consider a diversity of agents’ reasonings about environmental strategies are developed. Second, formulas for calculating equilibria in the games with three agents for arbitrary reflexion rank are derived.

Evolution of the acoustic startle response of Mexican cavefish

The ability to detect threatening sensory stimuli and initiate an escape response is essential for survival and under stringent evolutionary pressure. In diverse fish species, acoustic stimuli activate Mauthner neurons, which initiate a stereotypical C-start escape response. This reflexive behavior is highly conserved across aquatic species and provides a model for investigating the neural mechanism underlying the evolution of escape behavior. Here, we define evolved differences in the C-start response between populations of the Mexican cavefish, Astyanax mexicanus. Cave populations of A. mexicanus inhabit in an environment devoid of light and macroscopic predation, resulting in evolved differences in diverse morphological and behavioral traits. We find that the C-start is present in multiple populations of cavefish and river-dwelling surface fish, but response kinematics and probability differ between populations. The Pachón population of cavefish have an increased response probability, a slower response and reduction of the maximum bend angle, revealing evolved differences between surface and cave populations. In two other independently evolved populations of cavefish, the response probability and the kinematics of the response differ from one another, as well as from surface fish, suggesting the independent evolution of differences in the C-start response. Investigation of surface-cave hybrids reveals a relationship between angular speed and peak angle, suggesting these two kinematic characteristics are related at the genetic or functional levels. Together, these findings provide support for the use of A. mexicanus as a model to investigate the evolution of escape behavior.

Information generation as a functional basis of consciousness

What is the biological advantage of having consciousness? Functions of consciousness have been elusive due to the subjective nature of consciousness and ample empirical evidence showing the presence of many nonconscious cognitive performances in the human brain. Drawing upon empirical literature, here, we propose that a core function of consciousness be the ability to internally generate representations of events possibly detached from the current sensory input. Such representations are constructed by generative models learned through sensory-motor interactions with the environment. We argue that the ability to generate information underlies a variety of cognitive functions associated with consciousness such as intention, imagination, planning, short-term memory, attention, curiosity, and creativity, all of which contribute to non-reflexive behavior. According to this view, consciousness emerged in evolution when organisms gained the ability to perform internal simulations using internal models, which endowed them with flexible intelligent behavior. To illustrate the notion of information generation, we take variational autoencoders (VAEs) as an analogy and show that information generation corresponds the decoding (or decompression) part of VAEs. In biological brains, we propose that information generation corresponds to top-down predictions in the predictive coding framework. This is compatible with empirical observations that recurrent feedback activations are linked with consciousness whereas feedforward processing alone seems to occur without evoking conscious experience. Taken together, the information generation hypothesis captures many aspects of existing ideas about potential functions of consciousness and provides new perspectives on the functional roles of consciousness.

Sun navigation requires compass neurons in Drosophila

To follow a straight course, animals must maintain a constant heading relative to a fixed, distant landmark, a strategy termed menotaxis. In experiments using a flight simulator, we found that Drosophila adopt arbitrary headings with respect to a simulated sun, and individuals remember their heading preference between successive flights—even over gaps lasting several hours. Imaging experiments revealed that a class of neurons within the central complex, which have been previously shown to act as an internal compass, track the azimuthal motion of a sun stimulus. When these neurons are silenced, flies no longer adopt and maintain arbitrary headings, but instead exhibit frontal phototaxis. Thus, without the compass system, flies lose the ability to execute menotaxis and revert to a simpler, reflexive behavior.One sentence summarySilencing the compass neurons in the central complex of Drosophila eliminates sun navigation but leaves phototaxis intact.

Gaze-stabilizing central vestibular neurons project asymmetrically to extraocular motoneuron pools

Within reflex circuits, specific anatomical projections allow central neurons to relay sensations to effectors that generate movements. A major challenge is to relate anatomical features of central neural populations — such as asymmetric connectivity — to the computations the populations perform. To address this problem, we mapped the anatomy, modeled the function, and discovered a new behavioral role for a genetically-defined population of central vestibular neurons in rhombomeres 5-7 of larval zebrafish. First, we found that neurons within this central population project preferentially to motoneurons that move the eyes downward. Concor-dantly, when the entire population of asymmetrically-projecting neurons was stimulated collectively, only downward eye rotations were observed, demonstrating a functional correlate of the anatomical bias. When these neurons are ablated, fish failed to rotate their eyes following either nose-up or nose-down body tilts. This asymmetrically-projecting central population thus participates in both up and downward gaze stabilization. In addition to projecting to motoneurons, central vestibular neurons also receive direct sensory input from peripheral afferents. To infer whether asymmetric projections can facilitate sensory encoding or motor output, we modeled differentially-projecting sets of central vestibular neurons. Whereas motor command strength was independent of projection allocation, asymmetric projections enabled more accurate representation of nose-up stimuli. The model shows how asymmetric connectivity could enhance the representation of imbalance during nose-up postures while preserving gaze-stabilization performance. Finally, we found that central vestibular neurons were necessary for a vital behavior requiring maintenance of a nose-up posture: swim bladder inflation. These observations suggest that asymmetric connectivity in the vestibular system facilitates representation of ethologically-relevant stimuli without compromising reflexive behavior.Significance StatementInterneuron populations use specific anatomical projections to transform sensations into reflexive actions. Here we examined how the anatomical composition of a genetically-defined population of balance interneurons in the larval zebrafish relates to the computations it performs. First, we found that the population of interneurons that stabilize gaze preferentially project to motoneurons that move the eyes downward. Next, we discovered through modeling that such projection patterns can enhance the encoding of nose-up sensations without compromising gaze stabilization. Finally we found that loss of these interneurons impairs a vital behavior, swim bladder inflation, that relies on maintaining a nose-up posture. These observations suggest that anatomical specialization permits neural circuits to represent relevant features of the environment without compromising behavior.