A custom-made AAV1 variant (AAV1-T593K) enables efficient transduction of Japanese quail neurons in vitro and in vivo.

The widespread use of rodents in neuroscience has prompted the development of optimized viral variants for transduction of brain cells, in vivo. However, many of the viruses developed are less efficient in other model organisms, with birds being among the most resistant to transduction by current viral tools. Resultantly, the use of genetically-encoded tools and methods in avian species is markedly lower than in rodents; likely holding the field back. We sought to bridge this gap by developing custom viruses towards the transduction of brain cells of the Japanese quail. We first develop a protocol for culturing primary neurons and glia from quail embryos, followed by characterization of cultures via immunostaining, single cell mRNA sequencing, patch clamp electrophysiology and calcium imaging. We then leveraged the cultures for the rapid screening of various viruses, only to find that all yielded poor to no infection of cells in vitro. However, few infected neurons were obtained by AAV1 and AAV2. Scrutiny of the sequence of the AAV receptor found in quails led us to rationally design a custom-made AAV variant (AAV1-T593K; AAV1*) that exhibits improved transduction efficiencies in vitro and in vivo (14- and five-fold, respectively). Together, we present unique culturing method, transcriptomic profiles of quail's brain cells and a custom-tailored AAV1 for transduction of quail neurons in vitro and in vivo.

Spatial coding in the hippocampus and hyperpallium of flying owls.

The elucidation of spatial coding in the hippocampus requires exploring diverse animal species. While robust place-cells are found in the mammalian hippocampus, much less is known about spatial coding in the hippocampus of birds. Here we used a wireless-electrophysiology system to record single neurons in the hippocampus and other two dorsal pallial structures from freely flying barn owls (Tyto alba), a central-place nocturnal predator species with excellent navigational abilities. The owl's 3D position was monitored while it flew between perches. We found place cells-neurons that fired when the owl flew through a spatially restricted region in at least one direction-as well as neurons that encoded the direction of flight, and neurons that represented the owl's perching position between flights. Many neurons encoded combinations of position, direction, and perching. Spatial coding was maintained stable and invariant to lighting conditions. Place cells were observed in owls performing two different types of flying tasks, highlighting the generality of the result. Place coding was found in the anterior hippocampus and in the posterior part of the hyperpallium apicale, and to a lesser extent in the visual Wulst. The finding of place-cells in flying owls suggests commonalities in spatial coding across mammals and birds.

Spatial cognition in birds.

Finding the kitchen refrigerator in the middle of a sleepless night or arriving at the home parking lot at the end of a daily drive are things that we mostly take for granted. However, such feats rely on complex brain computations that integrate multiple environmental and internal cues. Together these computations mediate the process referred to as spatial cognition, the study of which has been one of the most active research fields in modern neuroscience. In the early 1970s, John O'Keefe recorded the activity of individual cells in the hippocampus of a freely moving rat: he found cells that fired action potentials when the rat passed through a specific place in the environment. Examining the activity pattern of such 'place cells', he concluded that the hippocampus may contain a 'cognitive map' that represents the animal's location in its proximate environment. Since O'Keefe's seminal discovery about half a century ago, extensive research has led to a detailed characterization of hippocampal space processing in rats and other mammalian species.

The Ecological View of Selective Attention.

Accumulating evidence is supporting the hypothesis that our selective attention is a manifestation of mechanisms that evolved early in evolution and are shared by many organisms from different taxa. This surge of new data calls for the re-examination of our notions about attention, which have been dominated mostly by human psychology. Here, we present an hypothesis that challenges, based on evolutionary grounds, a common view of attention as a means to manage limited brain resources. We begin by arguing that evolutionary considerations do not favor the basic proposition of the limited brain resources view of attention, namely, that the capacity of the sensory organs to provide information exceeds the capacity of the brain to process this information. Moreover, physiological studies in animals and humans show that mechanisms of selective attention are highly demanding of brain resources, making it paradoxical to see attention as a means to release brain resources. Next, we build on the above arguments to address the question why attention evolved in evolution. We hypothesize that, to a certain extent, limiting sensory processing is adaptive irrespective of brain capacity. We call this hypothesis the ecological view of attention (EVA) because it is centered on interactions of an animal with its environment rather than on internal brain resources. In its essence is the notion that inherently noisy and degraded sensory inputs serve the animal's adaptive, dynamic interactions with its environment. Attention primarily functions to resolve behavioral conflicts and false distractions. Hence, we evolved to focus on a particular target at the expense of others, not because of internal limitations, but to ensure that behavior is properly oriented and committed to its goals. Here, we expand on this notion and review evidence supporting it. We show how common results in human psychophysics and physiology can be reconciled with an EVA and discuss possible implications of the notion for interpreting current results and guiding future research.

Autonomous Flying With Neuromorphic Sensing.

Autonomous flight for large aircraft appears to be within our reach. However, launching autonomous systems for everyday missions still requires an immense interdisciplinary research effort supported by pointed policies and funding. We believe that concerted endeavors in the fields of neuroscience, mathematics, sensor physics, robotics, and computer science are needed to address remaining crucial scientific challenges. In this paper, we argue for a bio-inspired approach to solve autonomous flying challenges, outline the frontier of sensing, data processing, and flight control within a neuromorphic paradigm, and chart directions of research needed to achieve operational capabilities comparable to those we observe in nature. One central problem of neuromorphic computing is learning. In biological systems, learning is achieved by adaptive and relativistic information acquisition characterized by near-continuous information retrieval with variable rates and sparsity. This results in both energy and computational resource savings being an inspiration for autonomous systems. We consider pertinent features of insect, bat and bird flight behavior as examples to address various vital aspects of autonomous flight. Insects exhibit sophisticated flight dynamics with comparatively reduced complexity of the brain. They represent excellent objects for the study of navigation and flight control. Bats and birds enable more complex models of attention and point to the importance of active sensing for conducting more complex missions. The implementation of neuromorphic paradigms for autonomous flight will require fundamental changes in both traditional hardware and software. We provide recommendations for sensor hardware and processing algorithm development to enable energy efficient and computationally effective flight control.

Directional tuning in the hippocampal formation of birds.

Birds strongly rely on spatial memory and navigation. Therefore, it is of utmost interest to reveal how space is represented in the avian brain. Here we used tetrodes to record neurons from the hippocampal formation of Japanese quails-a ground-dwelling species-while the quails roamed in an open-field arena. Whereas spatially modulated cells (place cells, grid cells, border cells) were generally not encountered, the firing rate of about 12% of the neurons was unimodally and significantly modulated by the head azimuth-i.e., these were head-direction cells (HD cells). Typically, HD cells were maximally active at one preferred direction and minimally at the opposite null direction, with preferred directions spanning all 360° across the population. The preferred direction was independent of the animal's position and speed and was stable during the recording session. The HD tuning was broader compared to that of HD cells in rodents, and most cells had non-zero baseline firing in all directions. However, similar to findings in rodents, the HD tuning usually rotated with the rotation of a salient visual cue in the arena. Thus, these findings support the existence of an allocentric HD representation in the quail hippocampal formation and provide the first demonstration of HD cells in birds.

Behavioral and neuronal study of inhibition of return in barn owls.

Inhibition of return (IOR) is the reduction of detection speed and/or detection accuracy of a target in a recently attended location. This phenomenon, which has been discovered and studied thoroughly in humans, is believed to reflect a brain mechanism for controlling the allocation of spatial attention in a manner that enhances efficient search. Findings showing that IOR is robust, apparent at a very early age and seemingly dependent on midbrain activity suggest that IOR is a universal attentional mechanism in vertebrates. However, studies in non-mammalian species are scarce. To explore this hypothesis comparatively, we tested for IOR in barn owls (Tyto alba) using the classical Posner cueing paradigm. Two barn owls were trained to initiate a trial by fixating on the center of a computer screen and then turning their gaze to the location of a target. A short, non-informative cue appeared before the target, either at a location predicting the target (valid) or a location not predicting the target (invalid). In one barn owl, the response times (RT) to the valid targets compared to the invalid targets shifted from facilitation (lower RTs) to inhibition (higher RTs) when increasing the time lag between the cue and the target. The second owl mostly failed to maintain fixation and responded to the cue before the target onset. However, when including in the analysis only the trials in which the owl maintained fixation, an inhibition in the valid trials could be detected. To search for the neural correlates of IOR, we recorded multiunit responses in the optic tectum (OT) of four head-fixed owls passively viewing a cueing paradigm as in the behavioral experiments. At short cue to target lags (<100 ms), neural responses to the target in the receptive field (RF) were usually enhanced if the cue appeared earlier inside the RF (valid) and were suppressed if the cue appeared earlier outside the RF (invalid). This was reversed at longer lags: neural responses were suppressed in the valid conditions and were unaffected in the invalid conditions. The findings support the notion that IOR is a basic mechanism in the evolution of vertebrate behavior and suggest that the effect appears as a result of the interaction between lateral and forward inhibition in the tectal circuitry.

Harm-Benefit Analysis May Not Be the Best Approach to Ensure Minimal Harms and Maximal Benefits of Animal Research-Alternatives Should Be Explored.

Using animals in scientific research is commonly justified on the utilitarian basis that the benefits of scientific progress to human health and society exceed by far the harm inflicted on animals. In an attempt to ensure that this is indeed the case for every research project, legislation and guidelines increasingly demand the application of harm-benefit analysis (HBA) as part of the approval process of animal research protocols. The ethical principle of HBA asserts that the costs of an action should be weighed against the expected benefits. Any action that may inflict harm can only be approved if it is associated with a greater benefit. This principle is intuitively appealing but how to use it as a practical rule for ethical decisions is a difficult question. The main difficulty is that the future benefits of most scientific research are unmeasurable, unpredictable and are not manifested at the level of the individual project. Applying HBA in such cases may impede scientific progress by inducing a bias against basic research. Moreover, it can lead to the toleration of unnecessary harm to animals in research. Given these caveats of HBA, I call policy-makers to reconsider the place of HBA in animal research. Instead, I support an alternative guideline which is based on replacing the HBA principle (that the expected benefits of the research must exceed the harms caused to the animals) with two independent but mutually necessary principles: (1) any research using an animal must carry a benefit for society and (2) the harm inflicted to an animal in an experiment must be minimal and scientifically justified. I argue that rigorous harm-analysis, which is not weighted against obscure benefits, can increase the over-all benefits of research while reducing the harms to animals.

Self-motion trajectories can facilitate orientation-based figure-ground segregation.

Segregation of objects from the background is a basic and essential property of the visual system. We studied the neural detection of objects defined by orientation difference from background in barn owls (Tyto alba). We presented wide-field displays of densely packed stripes with a dominant orientation. Visual objects were created by orienting a circular patch differently from the background. In head-fixed conditions, neurons in both tecto- and thalamofugal visual pathways (optic tectum and visual Wulst) were weakly responsive to these objects in their receptive fields. However, notably, in freely viewing conditions, barn owls occasionally perform peculiar side-to-side head motions (peering) when scanning the environment. In the second part of the study we thus recorded the neural response from head-fixed owls while the visual displays replicated the peering conditions; i.e., the displays (objects and backgrounds) were shifted along trajectories that induced a retinal motion identical to sampled peering motions during viewing of a static object. These conditions induced dramatic neural responses to the objects, in the very same neurons that where unresponsive to the objects in static displays. By reverting to circular motions of the display, we show that the pattern of the neural response is mostly shaped by the orientation of the background relative to motion and not the orientation of the object. Thus our findings provide evidence that peering and/or other self-motions can facilitate orientation-based figure-ground segregation through interaction with inhibition from the surround.NEW & NOTEWORTHY Animals frequently move their sensory organs and thereby create motion cues that can enhance object segregation from background. We address a special example of such active sensing, in barn owls. When scanning the environment, barn owls occasionally perform small-amplitude side-to-side head movements called peering. We show that the visual outcome of such peering movements elicit neural detection of objects that are rotated from the dominant orientation of the background scene and which are otherwise mostly undetected. These results suggest a novel role for self-motions in sensing objects that break the regular orientation of elements in the scene.

Behavioral Evidence and Neural Correlates of Perceptual Grouping by Motion in the Barn Owl.

Perceiving an object as salient from its surround often requires a preceding process of grouping the object and background elements as perceptual wholes. In humans, motion homogeneity provides a strong cue for grouping, yet it is unknown to what extent this occurs in nonprimate species. To explore this question, we studied the effects of visual motion homogeneity in barn owls of both genders, at the behavioral as well as the neural level. Our data show that the coherency of the background motion modulates the perceived saliency of the target object. An object moving in an odd direction relative to other objects attracted more attention when the other objects moved homogeneously compared with when moved in a variety of directions. A possible neural correlate of this effect may arise in the population activity of the intermediate/deep layers of the optic tectum. In these layers, the neural responses to a moving element in the receptive field were suppressed when additional elements moved in the surround. However, when the surrounding elements all moved in one direction (homogeneously moving), they induced less suppression of the response compared with nonhomogeneously moving elements. Moreover, neural responses were more sensitive to the homogeneity of the background motion than to motion-direction contrasts between the receptive field and the surround. The findings suggest similar principles of saliency-by-motion in an avian species as in humans and show a locus in the optic tectum where the underlying neural circuitry may exist.SIGNIFICANCE STATEMENT A critical task of the visual system is to arrange incoming visual information to a meaningful scene of objects and background. In humans, elements that move homogeneously are grouped perceptually to form a categorical whole object. We discovered a similar principle in the barn owl's visual system, whereby the homogeneity of the motion of elements in the scene allows perceptually distinguishing an object from its surround. The novel findings of these visual effects in an avian species, which lacks neocortical structure, suggest that our basic visual perception shares more universal principles across species than presently thought, and shed light on possible brain mechanisms for perceptual grouping.

Interactions between top-down and bottom-up attention in barn owls (Tyto alba).

Selective attention, the prioritization of behaviorally relevant stimuli for behavioral control, is commonly divided into two processes: bottom-up, stimulus-driven selection and top-down, task-driven selection. Here, we tested two barn owls in a visual search task that examines attentional capture of the top-down task by bottom-up mechanisms. We trained barn owls to search for a vertical Gabor patch embedded in a circular array of differently oriented Gabor distractors (top-down guided search). To track the point of gaze, a lightweight wireless video camera was mounted on the owl's head. Three experiments were conducted in which the owls were tested in the following conditions: (1) five distractors; (2) nine distractors; (3) five distractors with one distractor surrounded by a red circle; or (4) five distractors with a brief sound at the initiation of the stimulus. Search times and number of head saccades to reach the target were measured and compared between the different conditions. It was found that search time and number of saccades to the target increased when the number of distractors was larger (condition 2) and when an additional irrelevant salient stimulus, auditory or visual, was added to the scene (conditions 3 and 4). These results demonstrate that in barn owls, bottom-up attention interacts with top-down attention to shape behavior in ways similar to human attentional capture. The findings suggest similar attentional principles in taxa that have been evolutionarily separated for 300 million years.

From biokinematics to a robotic active vision system.

Barn owls move their heads in very particular motions, compensating for the quasi-immovability of their eyes. These efficient predators often perform peering side-to-side head motions when scanning their surroundings and seeking prey. In this work, we use the head movements of barn owls as a model to bridge between biological active vision and machine vision. The biomotions are measured and used to actuate a specially built robot equipped with a depth camera for scanning. We hypothesize that the biomotions improve scan accuracy of static objects. Our experiments show that barn owl biomotion-based trajectories consistently improve scan accuracy when compared to intuitive scanning motions. This constitutes proof-of-concept evidence that the vision of robotic systems can be enhanced by bio-inspired viewpoint manipulation. Such biomimetic scanning systems can have many applications, e.g. manufacturing inspection or in autonomous robots.

The Neuroethological Paradox of Animal Consciousness.

The more advanced our understanding of the brain of an animal is, the less likely that this animal is a conscious being. This provocative logical paradox is explained and analyzed, leading to the conclusion that to advance understanding of animal consciousness it is necessary to resolve first how our consciousness is produced by our brain.

Stimulus-specific adaptation to visual but not auditory motion direction in the barn owl's optic tectum.

Whether the auditory and visual systems use a similar coding strategy to represent motion direction is an open question. We investigated this question in the barn owl's optic tectum (OT) testing stimulus-specific adaptation (SSA) to the direction of motion. SSA, the reduction of the response to a repetitive stimulus that does not generalize to other stimuli, has been well established in OT neurons. SSA suggests a separate representation of the adapted stimulus in upstream pathways. So far, only SSA to static stimuli has been studied in the OT. Here, we examined adaptation to moving auditory and visual stimuli. SSA to motion direction was examined using repeated presentations of moving stimuli, occasionally switching motion to the opposite direction. Acoustic motion was either mimicked by varying binaural spatial cues or implemented in free field using a speaker array. While OT neurons displayed SSA to motion direction in visual space, neither stimulation paradigms elicited significant SSA to auditory motion direction. These findings show a qualitative difference in how auditory and visual motion is processed in the OT and support the existence of dedicated circuitry for representing motion direction in the early stages of visual but not the auditory system.

Responses to Pop-Out Stimuli in the Barn Owl's Optic Tectum Can Emerge through Stimulus-Specific Adaptation.

UNLABELLED: Here, we studied neural correlates of orientation-contrast-based saliency in the optic tectum (OT) of barn owls. Neural responses in the intermediate/deep layers of the OT were recorded from lightly anesthetized owls confronted with arrays of bars in which one bar (the target) was orthogonal to the remaining bars (the distractors). Responses to target bars were compared with responses to distractor bars in the receptive field (RF). Initially, no orientation-contrast sensitivity was observed. However, if the position of the target bar in the array was randomly shuffled across trials so that it occasionally appeared in the RF, then such sensitivity emerged. The effect started to become significant after three or four positional changes of the target bar and strengthened with additional trials. Our data further suggest that this effect arises due to specific adaptation to the stimulus in the RF combined with suppression from the surround. By jittering the position of the bar inside the RF across trials, we demonstrate that the adaptation has two components, one position specific and one orientation specific. The findings give rise to the hypothesis that barn owls, by active scanning of the scene, can induce adaptation of the tectal circuitry to the common orientation and thus achieve a "pop-out" of rare orientations. Such a model is consistent with several behavioral observations in owls and may be relevant to other visual features and species. SIGNIFICANCE STATEMENT: Natural scenes are often characterized by a dominant orientation, such as the scenery of a pine forest or the sand dunes in a windy desert. Therefore, orientation that contrasts the regularity of the scene is perceived salient for many animals as a means to break camouflage. By actively moving the scene between each trial, we show here that neurons in the retinotopic map of the barn owl's optic tectum specifically adapt to the common orientation, giving rise to preferential representation of odd orientations. Based on this, we suggest a new mechanism for orientation-based camouflage breaking that links active scanning of scenes with neural adaptation. This mechanism may be relevant to pop-out in other species and visual features.

Visual-auditory integration for visual search: a behavioral study in barn owls.

Barn owls are nocturnal predators that rely on both vision and hearing for survival. The optic tectum of barn owls, a midbrain structure involved in selective attention, has been used as a model for studying visual-auditory integration at the neuronal level. However, behavioral data on visual-auditory integration in barn owls are lacking. The goal of this study was to examine if the integration of visual and auditory signals contributes to the process of guiding attention toward salient stimuli. We attached miniature wireless video cameras on barn owls' heads (OwlCam) to track their target of gaze. We first provide evidence that the area centralis (a retinal area with a maximal density of photoreceptors) is used as a functional fovea in barn owls. Thus, by mapping the projection of the area centralis on the OwlCam's video frame, it is possible to extract the target of gaze. For the experiment, owls were positioned on a high perch and four food items were scattered in a large arena on the floor. In addition, a hidden loudspeaker was positioned in the arena. The positions of the food items and speaker were changed every session. Video sequences from the OwlCam were saved for offline analysis while the owls spontaneously scanned the room and the food items with abrupt gaze shifts (head saccades). From time to time during the experiment, a brief sound was emitted from the speaker. The fixation points immediately following the sounds were extracted and the distances between the gaze position and the nearest items and loudspeaker were measured. The head saccades were rarely toward the location of the sound source but to salient visual features in the room, such as the door knob or the food items. However, among the food items, the one closest to the loudspeaker had the highest probability of attracting a gaze shift. This result supports the notion that auditory signals are integrated with visual information for the selection of the next visual search target.

Coding space-time stimulus dynamics in auditory brain maps.

Sensory maps are often distorted representations of the environment, where ethologically-important ranges are magnified. The implication of a biased representation extends beyond increased acuity for having more neurons dedicated to a certain range. Because neurons are functionally interconnected, non-uniform representations influence the processing of high-order features that rely on comparison across areas of the map. Among these features are time-dependent changes of the auditory scene generated by moving objects. How sensory representation affects high order processing can be approached in the map of auditory space of the owl's midbrain, where locations in the front are over-represented. In this map, neurons are selective not only to location but also to location over time. The tuning to space over time leads to direction selectivity, which is also topographically organized. Across the population, neurons tuned to peripheral space are more selective to sounds moving into the front. The distribution of direction selectivity can be explained by spatial and temporal integration on the non-uniform map of space. Thus, the representation of space can induce biased computation of a second-order stimulus feature. This phenomenon is likely observed in other sensory maps and may be relevant for behavior.

New perspectives on the owl's map of auditory space.

A map of sound direction was found in the owl's midbrain more than three decades ago. This finding suggested that the brain reconstructs spatial coordinates to represent them. Subsequent research elucidated the variables used to compute the map. Here we provide a review of the processes leading to its emergence and an updated perspective on how and what information is represented.

Saliency mapping in the optic tectum and its relationship to habituation.

Habituation of the orienting response has long served as a model system for studying fundamental psychological phenomena such as learning, attention, decisions, and surprise. In this article, we review an emerging hypothesis that the evolutionary role of the superior colliculus (SC) in mammals or its homolog in birds, the optic tectum (OT), is to select the most salient target and send this information to the appropriate brain regions to control the body and brain orienting responses. Recent studies have begun to reveal mechanisms of how saliency is computed in the OT/SC, demonstrating a striking similarity between mammals and birds. The saliency of a target can be determined by how different it is from the surrounding objects, by how different it is from its history (that is habituation) and by how relevant it is for the task at hand. Here, we will first review evidence, mostly from primates and barn owls, that all three types of saliency computations are linked in the OT/SC. We will then focus more on neural adaptation in the OT and its possible link to temporal saliency and habituation.