Walking bumblebees see faster.

The behavioural state of animals has profound effects on neuronal information processing. Locomotion changes the response properties of visual interneurons in the insect brain, but it is still unknown if it also alters the response properties of photoreceptors. Photoreceptor responses become faster at higher temperatures. It has therefore been suggested that thermoregulation in insects could improve temporal resolution in vision, but direct evidence for this idea has so far been missing. Here, we compared electroretinograms from the compound eyes of tethered bumblebees that were either sitting or walking on an air-supported ball. We found that the visual processing speed strongly increased when the bumblebees were walking. By monitoring the eye temperature during recording, we saw that the increase in response speed was in synchrony with a rise in eye temperature. By artificially heating the head, we show that the walking-induced temperature increase of the visual system is sufficient to explain the rise in processing speed. We also show that walking accelerates the visual system to the equivalent of a 14-fold increase in light intensity. We conclude that the walking-induced rise in temperature accelerates the processing of visual information-an ideal strategy to process the increased information flow during locomotion.

Unraveling the neural basis of spatial orientation in arthropods.

The neural basis underlying spatial orientation in arthropods, in particular insects, has received considerable interest in recent years. This special issue of the Journal of Comparative Physiology A seeks to take account of these developments by presenting a collection of eight review articles and eight original research articles highlighting hotspots of research on spatial orientation in arthropods ranging from flies to spiders and the underlying neural circuits. The contributions impressively illustrate the wide range of tools available to arthropods extending from specific sensory channels to highly sophisticated neural computations for mastering complex navigational challenges.

The sky compass network in the brain of the desert locust.

Many arthropods and vertebrates use celestial signals such as the position of the sun during the day or stars at night as compass cues for spatial orientation. The neural network underlying sky compass coding in the brain has been studied in great detail in the desert locust Schistocerca gregaria. These insects perform long-range migrations in Northern Africa and the Middle East following seasonal changes in rainfall. Highly specialized photoreceptors in a dorsal rim area of their compound eyes are sensitive to the polarization of the sky, generated by scattered sunlight. These signals are combined with direct information on the sun position in the optic lobe and anterior optic tubercle and converge from both eyes in a midline crossing brain structure, the central complex. Here, head direction coding is achieved by a compass-like arrangement of columns signaling solar azimuth through a 360° range of space by combining direct brightness cues from the sun with polarization cues matching the polarization pattern of the sky. Other directional cues derived from wind direction and internal self-rotation input are likely integrated. Signals are transmitted as coherent steering commands to descending neurons for directional control of locomotion and flight.

Matched-filter coding of sky polarization results in an internal sun compass in the brain of the desert locust.

Many animals use celestial cues for spatial orientation. These include the sun and, in insects, the polarization pattern of the sky, which depends on the position of the sun. The central complex in the insect brain plays a key role in spatial orientation. In desert locusts, the angle of polarized light in the zenith above the animal and the direction of a simulated sun are represented in a compass-like fashion in the central complex, but how both compasses fit together for a unified representation of external space remained unclear. To address this question, we analyzed the sensitivity of intracellularly recorded central-complex neurons to the angle of polarized light presented from up to 33 positions in the animal's dorsal visual field and injected Neurobiotin tracer for cell identification. Neurons were polarization sensitive in large parts of the virtual sky that in some cells extended to the horizon in all directions. Neurons, moreover, were tuned to spatial patterns of polarization angles that matched the sky polarization pattern of particular sun positions. The horizontal components of these calculated solar positions were topographically encoded in the protocerebral bridge of the central complex covering 360° of space. This whole-sky polarization compass does not support the earlier reported polarization compass based on stimulation from a small spot above the animal but coincides well with the previously demonstrated direct sun compass based on unpolarized light stimulation. Therefore, direct sunlight and whole-sky polarization complement each other for robust head direction coding in the locust central complex.

Rotation of skylight polarization during learning walks is necessary to trigger neuronal plasticity in Cataglyphis ants.

Many animals use celestial cues for impressive navigational performances in challenging habitats. Since the position of the sun and associated skylight cues change throughout the day and season, it is crucial to correct for these changes. Cataglyphis desert ants possess a time-compensated skylight compass allowing them to navigate back to their nest using the shortest way possible. The ants have to learn the sun's daily course (solar ephemeris) during initial learning walks (LW) before foraging. This learning phase is associated with substantial structural changes in visual neuronal circuits of the ant's brain. Here, we test whether the rotation of skylight polarization during LWs is the necessary cue to induce learning-dependent rewiring in synaptic circuits in high-order integration centres of the ant brain. Our results show that structural neuronal changes in the central complex and mushroom bodies are triggered only when LWs were performed under a rotating skylight polarization pattern. By contrast, when naive ants did not perform LWs, but were exposed to skylight cues, plasticity was restricted to light spectrum-dependent changes in synaptic complexes of the lateral complex. The results identify sky-compass cues triggering learning-dependent versus -independent neuronal plasticity during the behavioural transition from interior workers to outdoor foragers.

Performance of polarization-sensitive neurons of the locust central complex at different degrees of polarization.

The polarization pattern of the sky is exploited by many insects for spatial orientation and navigation. It derives from Rayleigh scattering in the atmosphere and depends directly on the position of the sun. In the insect brain, the central complex (CX) houses neurons tuned to the angle of polarization (AoP), that together constitute an internal compass for celestial navigation. Polarized light is not only characterized by the AoP, but also by the degree of polarization (DoP), which can be highly variable, depending on sky conditions. Under a clear sky, the DoP of polarized sky light may reach up to 0.75 but is usually much lower especially when light is scattered by clouds or haze. To investigate how the polarization-processing network of the CX copes with low DoPs, we recorded intracellularly from neurons of the locust CX at different stages of processing, while stimulating with light of different DoPs. Significant responses to polarized light occurred down to DoPs of 0.05 indicating reliable coding of the AoP even at unfavorable sky conditions. Moreover, we found that the activity of neurons at the CX input stage may be strongly influenced by nearly unpolarized light, while the activity of downstream neurons appears less affected.

Stimulus-dependent orientation strategies in monarch butterflies.

Insects are well known for their ability to keep track of their heading direction based on a combination of skylight cues and visual landmarks. This allows them to navigate back to their nest, disperse throughout unfamiliar environments, as well as migrate over large distances between their breeding and non-breeding habitats. The monarch butterfly (Danaus plexippus), for instance, is known for its annual southward migration from North America to certain trees in Central Mexico. To maintain a constant flight route, these butterflies use a time-compensated sun compass, which is processed in a region in the brain, termed the central complex. However, to successfully complete their journey, the butterflies' brain must generate a multitude of orientation strategies, allowing them to dynamically switch from sun-compass orientation to a tactic behavior toward a certain target. To study whether monarch butterflies exhibit different orientation modes and if they can switch between them, we observed the orientation behavior of tethered flying butterflies in a flight simulator while presenting different visual cues to them. We found that the butterflies' behavior depended on the presented visual stimulus. Thus, while a dark stripe was used for flight stabilization, a bright stripe was fixated by the butterflies in their frontal visual field. If we replaced a bright stripe with a simulated sun stimulus, the butterflies switched their behavior and exhibited compass orientation. Taken together, our data show that monarch butterflies rely on and switch between different orientation modes, allowing the animal to adjust orientation to its actual behavioral demands.

A micro-CT-based standard brain atlas of the bumblebee.

In recent years, bumblebees have become a prominent insect model organism for a variety of biological disciplines, particularly to investigate learning behaviors as well as visual performance. Understanding these behaviors and their underlying neurobiological principles requires a clear understanding of brain anatomy. Furthermore, to be able to compare neuronal branching patterns across individuals, a common framework is required, which has led to the development of 3D standard brain atlases in most of the neurobiological insect model species. Yet, no bumblebee 3D standard brain atlas has been generated. Here we present a brain atlas for the buff-tailed bumblebee Bombus terrestris using micro-computed tomography (micro-CT) scans as a source for the raw data sets, rather than traditional confocal microscopy, to produce the first ever micro-CT-based insect brain atlas. We illustrate the advantages of the micro-CT technique, namely, identical native resolution in the three cardinal planes and 3D structure being better preserved. Our Bombus terrestris brain atlas consists of 30 neuropils reconstructed from ten individual worker bees, with micro-CT allowing us to segment neuropils completely intact, including the lamina, which is a tissue structure often damaged when dissecting for immunolabeling. Our brain atlas can serve as a platform to facilitate future neuroscience studies in bumblebees and illustrates the advantages of micro-CT for specific applications in insect neuroanatomy.

Two Compasses in the Central Complex of the Locust Brain.

Many migratory insects rely on a celestial compass for spatial orientation. Several features of the daytime sky, all generated by the sun, can be exploited for navigation. Two of these are the position of the sun and the pattern of polarized skylight. Neurons of the central complex (CX), a group of neuropils in the central brain of insects, have been shown to encode sky compass cues. In desert locusts, the CX holds a topographic, compass-like representation of the plane of polarized light (E-vector) presented from dorsal direction. In addition, these neurons also encode the azimuth of an unpolarized light spot, likely representing the sun. Here, we investigate whether, in addition to E-vector orientation, the solar azimuth is represented topographically in the CX. We recorded intracellularly from eight types of CX neuron while stimulating animals of either sex with polarized blue light from zenithal direction and an unpolarized green light spot rotating around the animal's head at different elevations. CX neurons did not code for elevation of the unpolarized light spot. However, two types of columnar neuron showed a linear correlation between innervated slice in the CX and azimuth tuning to the unpolarized green light spot, consistent with an internal compass representation of solar azimuth. Columnar outputs of the CX also showed a topographic representation of zenithal E-vector orientation, but the two compasses were not linked to each other. Combined stimulation with unpolarized green and polarized blue light suggested that the two compasses interact in a nonlinear way.SIGNIFICANCE STATEMENT In the brain of the desert locust, neurons sensitive to the plane of celestial polarization are arranged like a compass in the slices of the central complex (CX). These neurons, in addition, code for the horizontal direction of an unpolarized light cue possibly representing the sun. We show here that horizontal directions are, in addition to E-vector orientations from the dorsal direction, represented in a compass-like manner across the slices of the CX. However, the two compasses are not linked to each other, but rather seem to interact in a cell-specific, nonlinear way. Our study confirms the role of the CX in signaling heading directions and shows that different cues are used for this task.

Spatial orientation based on multiple visual cues in non-migratory monarch butterflies.

Monarch butterflies (Danaus plexippus) are prominent for their annual long-distance migration from North America to their overwintering area in Central Mexico. To find their way on this long journey, they use a sun compass as their main orientation reference but will also adjust their migratory direction with respect to mountain ranges. This indicates that the migratory butterflies also attend to the panorama to guide their travels. Although the compass has been studied in detail in migrating butterflies, little is known about the orientation abilities of non-migrating butterflies. Here, we investigated whether non-migrating butterflies - which stay in a more restricted area to feed and breed - also use a similar compass system to guide their flights. Performing behavioral experiments on tethered flying butterflies in an indoor LED flight simulator, we found that the monarchs fly along straight tracks with respect to a simulated sun. When a panoramic skyline was presented as the only orientation cue, the butterflies maintained their flight direction only during short sequences, suggesting that they potentially use it for flight stabilization. We further found that when we presented the two cues together, the butterflies incorporate both cues in their compass. Taken together, we show here that non-migrating monarch butterflies can combine multiple visual cues for robust orientation, an ability that may also aid them during their migration.

The role of lateral optic flow cues in hawkmoth flight control.

Flying animals require sensory feedback on changes of their body position, as well as on their distance from nearby objects. The apparent image motion, or optic flow, which is generated as animals move through the air, can provide this information. Flight tunnel experiments have been crucial for our understanding of how insects use optic flow for flight control in confined spaces. However, previous work mainly focused on species from two insect orders: Hymenoptera and Diptera. We therefore set out to investigate whether the previously described control strategies to navigate enclosed environments are also used by insects with a different optical system, flight kinematics and phylogenetic background. We tested the role of lateral visual cues for forward flight control in the hummingbird hawkmoth Macroglossum stellatarum (Sphingidae, Lepidoptera), which possesses superposition compound eyes, and has the ability to hover in addition to its capacity for fast forward flight. Our results show that hawkmoths use a similar strategy for lateral position control to bees and flies in balancing the magnitude of translational optic flow perceived in both eyes. However, the influence of lateral optic flow on flight speed in hawkmoths differed from that in bees and flies. Moreover, hawkmoths showed individually attributable differences in position and speed control when the presented optic flow was unbalanced.

Integration of celestial compass cues in the central complex of the locust brain.

Many insects rely on celestial compass cues such as the polarization pattern of the sky for spatial orientation. In the desert locust, the central complex (CX) houses multiple sets of neurons, sensitive to the oscillation plane of polarized light and thus probably acts as an internal polarization compass. We investigated whether other sky compass cues like direct sunlight or the chromatic gradient of the sky might contribute to this compass. We recorded from polarization-sensitive CX neurons while an unpolarized green or ultraviolet light spot was moved around the head of the animal. All types of neuron that were sensitive to the plane of polarization (E-vector) above the animal also responded to the unpolarized light spots in an azimuth-dependent way. The tuning to the unpolarized light spots was independent of wavelength, suggesting that the neurons encode solar azimuth based on direct sunlight and not on the sky chromatic gradient. Two cell types represented the natural 90 deg relationship between solar azimuth and zenithal E-vector orientation, providing evidence to suggest that solar azimuth information supports the internal polarization compass. Most neurons showed advances in their tuning to the E-vector and the unpolarized light spots dependent on rotation direction, consistent with anticipatory signaling. The amplitude of responses and its variability were dependent on the level of background firing, possibly indicating different internal states. The integration of polarization and solar azimuth information strongly suggests that besides the polarization pattern of the sky, direct sunlight might be an important cue for sky compass navigation in the locust.

Nonlinearization: naturalistic stimulation and nonlinear dynamic behavior in a spider mechanoreceptor.

In a previous study, we used linear frequency response analysis to show that naturalistic stimulation of spider primary mechanosensory neurons produced different response dynamics than the commonly used Gaussian random noise. We isolated this difference to the production of action potentials from receptor potential and suggested that the different distribution of frequency components in the naturalistic signal increased the nonlinearity of action potential encoding. Here, we tested the relative contributions of first- and second-order processes to the action potential signal by measuring linear and quadratic coherence functions. Naturalistic stimulation shifted the linear coherence toward lower frequencies, while quadratic coherence was always higher than linear coherence and increased with naturalistic stimulation. In an initial attempt to separate the order of time-dependent and nonlinear processes, we fitted quadratic frequency response functions by two block-structured models consisting of a power-law filter and a static second-order nonlinearity in alternate cascade orders. The same cascade models were then fitted to the original time domain data by conventional numerical analysis algorithms, using a polynomial function as the static nonlinearity. Quadratic models with a linear filter followed by a static nonlinearity were favored over the reverse order, but with weak significance. Polynomial nonlinear functions indicated that rectification is a major nonlinearity. A complete quantitative description of sensory encoding in these primary mechanoreceptors remains elusive but clearly requires quadratic and higher nonlinear operations on the input signal to explain the sensitivity of dynamic behavior to different input signal patterns.

Neurons in the brain of the desert locust Schistocerca gregaria sensitive to polarized light at low stimulus elevations.

Desert locusts (Schistocerca gregaria) sense the plane of dorsally presented polarized light through specialized dorsal eye regions that are likely adapted to exploit the polarization pattern of the blue sky for spatial orientation. Receptive fields of these dorsal rim photoreceptors and polarization-sensitive interneurons are directed toward the upper sky but may extend to elevations below 30°. Behavioral data, however, suggests that S. gregaria is even able to detect polarized light from ventral directions but physiological evidence for this is still lacking. In this study we characterized neurons in the locust brain showing polarization sensitivity at low elevations down to the horizon. In most neurons polarization sensitivity was absent or weak when stimulating from the zenith. All neurons, including projection and commissural neurons of the optic lobe and local interneurons of the central brain, are novel cell types, distinct from polarization-sensitive neurons studied so far. Painting dorsal rim areas in both eyes black to block visual input had no effect on the polarization sensitivity of these neurons, suggesting that they receive polarized light input from the main eye. A possible role of these neurons in flight stabilization or the perception of polarized light reflected from bodies of water or vegetation is discussed.

Organization and functional roles of the central complex in the insect brain.

The central complex is a group of modular neuropils across the midline of the insect brain. Hallmarks of its anatomical organization are discrete layers, an organization into arrays of 16 slices along the right-left axis, and precise inter-hemispheric connections via chiasmata. The central complex is connected most prominently with the adjacent lateral complex and the superior protocerebrum. Its developmental appearance corresponds with the appearance of compound eyes and walking legs. Distinct dopaminergic neurons control various forms of arousal. Electrophysiological studies provide evidence for roles in polarized light vision, sky compass orientation, and integration of spatial information for locomotor control. Behavioral studies on mutant and transgenic flies indicate roles in spatial representation of visual cues, spatial visual memory, directional control of walking and flight, and place learning. The data suggest that spatial azimuthal directions (i.e., where) are represented in the slices, and cue information (i.e., what) are represented in different layers of the central complex.

Integration of polarization and chromatic cues in the insect sky compass.

Animals relying on a celestial compass for spatial orientation may use the position of the sun, the chromatic or intensity gradient of the sky, the polarization pattern of the sky, or a combination of these cues as compass signals. Behavioral experiments in bees and ants, indeed, showed that direct sunlight and sky polarization play a role in sky compass orientation, but the relative importance of these cues are species-specific. Intracellular recordings from polarization-sensitive interneurons in the desert locust and monarch butterfly suggest that inputs from different eye regions, including polarized-light input through the dorsal rim area of the eye and chromatic/intensity gradient input from the main eye, are combined at the level of the medulla to create a robust compass signal. Conflicting input from the polarization and chromatic/intensity channel, resulting from eccentric receptive fields, is eliminated at the level of the anterior optic tubercle and central complex through internal compensation for changing solar elevations, which requires input from a circadian clock. Across several species, the central complex likely serves as an internal sky compass, combining E-vector information with other celestial cues. Descending neurons, likewise, respond both to zenithal polarization and to unpolarized cues in an azimuth-dependent way.

The neuronal building blocks of the navigational toolkit in the central complex of insects.

The central complex in the brain of insects is a group of midline-spanning neuropils at the interface between sensory and premotor tasks of the brain. It is involved in sleep control, decision-making and most prominently in goal-directed locomotion behaviors. The recently published connectome of the central complex of Drosophila melanogaster is a milestone in understanding the intricacies of the central-complex circuits and will provide inspiration for testable hypotheses for the coming years. Here, I provide a basic neuroanatomical description of the central complex of Drosophila and other species and discuss some recent advancements, some of which, such as the discovery of coordinate transformation through vector math, have been predicted from connectomics data.

3D atlas of cerebral neuropils with previously unknown demarcations in the honey bee brain.

Honey bees (Apis mellifera) express remarkable social interactions and cognitive capabilities that have been studied extensively. In many cases, behavioral studies were accompanied by neurophysiological and neuroanatomical investigations. While most studies have focused on primary sensory neuropils, such as the optic lobes or antennal lobes, and major integration centers, such as the mushroom bodies or the central complex, many regions of the cerebrum (the central brain without the optic lobes) of the honey bee are only poorly explored so far, both anatomically and physiologically. To promote studies of these brain regions, we used anti-synapsin immunolabeling and neuronal tract tracings followed by confocal imaging and 3D reconstructions to demarcate all neuropils in the honey bee cerebrum and close this gap at the anatomical level. We demarcated 35 neuropils and 25 fiber tracts in the honey bee cerebrum, most of which have counterparts in the fly (Drosophila melanogaster) and other insect species that have been investigated so far at this level of detail. We discuss the role of cerebral neuropils in multisensory integration in the insect brain, emphasize the importance of this brain atlas for comparative studies, and highlight specific architectural features of the honey bee cerebrum.

Activation of GABAA receptors modulates all stages of mechanoreception in spider mechanosensory neurons.

GABA(A) receptors mediate mainly inhibitory effects, but there are also many examples of excitatory effects in both mammalian and invertebrate preparations. Here, we aimed to create a complete, quantitative picture of GABA(A)-mediated excitation in a mechanosensory neuron where this phenomenon has been well established. We used muscimol to activate GABA(A) receptors in spider VS-3 neurons and measured the dynamic behavior independently and separately at each of three stages of mechanoreception (receptor current, receptor potential, and action potentials) before and during modulation. We calculated frequency response functions between each stage, estimated information as signal entropy, and estimated information capacity from coherence. Since coherence is sensitive to both noise and nonlinearity, we measured signal-to-noise separately at each stage by averaging responses to repeated mechanical inputs. Muscimol depolarized VS-3 neurons and, after brief inhibition, increased their firing rates. During this excitation, we found significant changes at each stage. Receptor current was attenuated but became more selective to high frequencies. Membrane impedance and time constant fell, favoring higher frequency transmission from receptor current to receptor potential. Action potential firing increased and had higher total entropy. Information capacity from signal-to-noise was always much higher than from coherence, confirming that intracellular noise does not limit signal transmission in these neurons. We conclude that GABA(A) receptor activation shifts each stage of mechanotransduction to higher frequency sensitivity, while the elevated firing rate increases the amount of information that can be encoded. These results show that a single neurotransmitter can finely modulate a sensory neuron's sensitivity and ability to transmit information.