Importance of magnetic information for neuronal plasticity in desert ants.

Many animal species rely on the Earth's magnetic field during navigation, but where in the brain magnetic information is processed is still unknown. To unravel this, we manipulated the natural magnetic field at the nest entrance of Cataglyphis desert ants and investigated how this affects relevant brain regions during early compass calibration. We found that manipulating the Earth's magnetic field has profound effects on neuronal plasticity in two sensory integration centers. Magnetic field manipulations interfere with a typical look-back behavior during learning walks of naive ants. Most importantly, structural analyses in the ants' neuronal compass (central complex) and memory centers (mushroom bodies) demonstrate that magnetic information affects neuronal plasticity during early visual learning. This suggests that magnetic information does not only serve as a compass cue for navigation but also as a global reference system crucial for spatial memory formation. We propose a neural circuit for integration of magnetic information into visual guidance networks in the ant brain. Taken together, our results provide an insight into the neural substrate for magnetic navigation in insects.

Expansion microscopy in honeybee brains for high-resolution neuroanatomical analyses in social insects.

The diffraction limit of light microscopy poses a problem that is frequently faced in structural analyses of social insect brains. With the introduction of expansion microscopy (ExM), a tool became available to overcome this limitation by isotropic physical expansion of preserved specimens. Our analyses focus on synaptic microcircuits (microglomeruli, MG) in the mushroom body (MB) of social insects, high-order brain centers for sensory integration, learning, and memory. MG undergo significant structural reorganizations with age, sensory experience, and during long-term memory formation. However, the changes in subcellular architecture involved in this plasticity have only partially been accessed yet. Using the western honeybee Apis mellifera as an experimental model, we established ExM for the first time in a social insect species and applied it to investigate plasticity in synaptic microcircuits within MG of the MB calyces. Using combinations of antibody staining and neuronal tracing, we demonstrate that this technique enables quantitative and qualitative analyses of structural neuronal plasticity at high resolution in a social insect brain.

The role of learning-walk related multisensory experience in rewiring visual circuits in the desert ant brain.

Efficient spatial orientation in the natural environment is crucial for the survival of most animal species. Cataglyphis desert ants possess excellent navigational skills. After far-ranging foraging excursions, the ants return to their inconspicuous nest entrance using celestial and panoramic cues. This review focuses on the question about how naïve ants acquire the necessary spatial information and adjust their visual compass systems. Naïve ants perform structured learning walks during their transition from the dark nest interior to foraging under bright sunlight. During initial learning walks, the ants perform rotational movements with nest-directed views using the earth's magnetic field as an earthbound compass reference. Experimental manipulations demonstrate that specific sky compass cues trigger structural neuronal plasticity in visual circuits to integration centers in the central complex and mushroom bodies. During learning walks, rotation of the sky-polarization pattern is required for an increase in volume and synaptic complexes in both integration centers. In contrast, passive light exposure triggers light-spectrum (especially UV light) dependent changes in synaptic complexes upstream of the central complex. We discuss a multisensory circuit model in the ant brain for pathways mediating structural neuroplasticity at different levels following passive light exposure and multisensory experience during the performance of learning walks.

Contact chemoreception, magnetic maps, thermoregulation by a superorganism, and, thanks to Einstein, an all-time record: the Editors' and Readers' Choice Awards 2023.

During the 99 years of its history, the Journal of Comparative Physiology A has published many of the most influential papers in comparative physiology and related disciplines. To celebrate this achievement of the journal's authors, annual Editors' Choice Awards and Readers' Choice Awards are presented. The winners of the 2023 Editors' Choice Awards are 'Contact chemoreception in multi­modal sensing of prey by Octopus' by Buresch et al. (J Comp Physiol A 208:435-442, 2022) in the Original Paper category; and 'Magnetic maps in animal navigation' by Lohmann et al. (J Comp Physiol A 208:41-67, 2022) in the Review/Review-History Article category. The winners of the 2023 Readers' Choice Awards are 'Coping with the cold and fighting the heat: thermal homeostasis of a superorganism, the honeybee colony' by Stabentheiner et al. (J Comp Physiol A 207:337-351; 2021) in the Original Paper category; and 'Einstein, von Frisch and the honeybee: a historical letter comes to light' by Dyer et al. (J Comp Physiol A 207:449-456, 2021) in the Review/Review-History category.

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.

Government funding of research beyond biomedicine: challenges and opportunities for neuroethology.

Curiosity-driven research is fundamental for neuroethology and depends crucially on governmental funding. Here, we highlight similarities and differences in funding of curiosity-driven research across countries by comparing two major funding agencies-the National Science Foundation (NSF) in the United States and the German Research Foundation (Deutsche Forschungsgemeinschaft, DFG). We interviewed representatives from each of the two agencies, focusing on general funding trends, levels of young investigator support, career-life balance, and international collaborations. While our analysis revealed a negative trend in NSF funding of biological research, including curiosity-driven research, German researchers in these areas have benefited from a robust positive trend in DFG funding. The main reason for the decrease in curiosity-driven research in the US is that the NSF has only partially been able to compensate for the funding gap resulting from the National Institutes of Health restricting their support to biomedical research using select model organisms. Notwithstanding some differences in funding programs, particularly those relevant for scientists in the postdoctoral phase, both the NSF and DFG clearly support curiosity-driven research.

Magnetosensation during re-learning walks in desert ants (Cataglyphis nodus).

At the beginning of their foraging careers, Cataglyphis desert ants calibrate their compass systems and learn the visual panorama surrounding the nest entrance. For that, they perform well-structured initial learning walks. During rotational body movements (pirouettes), naïve ants (novices) gaze back to the nest entrance to memorize their way back to the nest. To align their gaze directions, they rely on the geomagnetic field as a compass cue. In contrast, experienced ants (foragers) use celestial compass cues for path integration during food search. If the panorama at the nest entrance is changed, foragers perform re-learning walks prior to heading out on new foraging excursions. Here, we show that initial learning walks and re-learning walks are structurally different. During re-learning walks, foragers circle around the nest entrance before leaving the nest area to search for food. During pirouettes, they do not gaze back to the nest entrance. In addition, foragers do not use the magnetic field as a compass cue to align their gaze directions during re-learning walk pirouettes. Nevertheless, magnetic alterations during re-learning walks under manipulated panoramic conditions induce changes in nest-directed views indicating that foragers are still magnetosensitive in a cue conflict situation.

It's all about seeing and hearing: the Editors' and Readers' Choice Awards 2022.

This year marks the inauguration of the annual Editors' Choice Award and the Readers' Choice Award, each presented for outstanding original papers and review articles published in the Journal of Comparative Physiology A. The winners of the 2022 Editors' Choice Award were determined by vote of the Editorial Board for the most highly recommended papers published in Volume 207 in 2021. They are 'Visual discrimination and resolution in freshwater stingrays (Potamotrygon motoro)' by Daniel et al. (J Comp Physiol A 207, 43-58, 2021) in the Original Paper category; and 'Neurophysiology goes wild: from exploring sensory coding in sound proof rooms to natural environments' by Römer (J Comp Physiol A 207, 303-319, 2021) in the Review Article category. The 2022 Readers' Choice Award was based on access number of articles published in Volume 206 in 2020, to ensure at least 12-month online presence. It is given to Nicholas et al. for their original paper titled 'Visual motion sensitivity in descending neurons in the hoverfly' (J Comp Physiol A 206, 149-163, 2020); and to Schnaitmann et al. for their review article entitled 'Color vision in insects: insights from Drosophila' (J Comp Physiol A 206, 183-198, 2020).

Transcriptomic, peptidomic, and mass spectrometry imaging analysis of the brain in the ant Cataglyphis nodus.

Behavioral flexibility is an important cornerstone for the ecological success of animals. Social Cataglyphis nodus ants with their age-related polyethism characterized by age-related behavioral phenotypes represent a prime example for behavioral flexibility. We propose neuropeptides as powerful candidates for the flexible modulation of age-related behavioral transitions in individual ants. As the neuropeptidome of C. nodus was unknown, we collected a comprehensive peptidomic data set obtained by transcriptome analysis of the ants' central nervous system combined with brain extract analysis by Q-Exactive Orbitrap mass spectrometry (MS) and direct tissue profiling of different regions of the brain by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. In total, we identified 71 peptides with likely bioactive function, encoded on 49 neuropeptide-, neuropeptide-like, and protein hormone prepropeptide genes, including a novel neuropeptide-like gene (fliktin). We next characterized the spatial distribution of a subset of peptides encoded on 16 precursor proteins with high resolution by MALDI MS imaging (MALDI MSI) on 14 µm brain sections. The accuracy of our MSI data were confirmed by matching the immunostaining patterns for tachykinins with MSI ion images from consecutive brain sections. Our data provide a solid framework for future research into spatially resolved qualitative and quantitative peptidomic changes associated with stage-specific behavioral transitions and the functional role of neuropeptides in Cataglyphis ants.

Plasticity and modulation of olfactory circuits in insects.

Olfactory circuits change structurally and physiologically during development and adult life. This allows insects to respond to olfactory cues in an appropriate and adaptive way according to their physiological and behavioral state, and to adapt to their specific abiotic and biotic natural environment. We highlight here findings on olfactory plasticity and modulation in various model and non-model insects with an emphasis on moths and social Hymenoptera. Different categories of plasticity occur in the olfactory systems of insects. One type relates to the reproductive or feeding state, as well as to adult age. Another type of plasticity is context-dependent and includes influences of the immediate sensory and abiotic environment, but also environmental conditions during postembryonic development, periods of adult behavioral maturation, and short- and long-term sensory experience. Finally, plasticity in olfactory circuits is linked to associative learning and memory formation. The vast majority of the available literature summarized here deals with plasticity in primary and secondary olfactory brain centers, but also peripheral modulation is treated. The described molecular, physiological, and structural neuronal changes occur under the influence of neuromodulators such as biogenic amines, neuropeptides, and hormones, but the mechanisms through which they act are only beginning to be analyzed.

Magnetoreception in Hymenoptera: importance for navigation.

The use of information provided by the geomagnetic field (GMF) for navigation is widespread across the animal kingdom. At the same time, the magnetic sense is one of the least understood senses. Here, we review evidence for magnetoreception in Hymenoptera. We focus on experiments aiming to shed light on the role of the GMF for navigation. Both honeybees and desert ants are well-studied experimental models for navigation, and both use the GMF for specific navigational tasks under certain conditions. Cataglyphis desert ants use the GMF as a compass cue for path integration during their initial learning walks to align their gaze directions towards the nest entrance. This represents the first example for the use of the GMF in an insect species for a genuine navigational task under natural conditions and with all other navigational cues available. We argue that the recently described magnetic compass in Cataglyphis opens up a new integrative approach to understand the mechanisms underlying magnetoreception in Hymenoptera on different biological levels.

Immediate early genes in social insects: a tool to identify brain regions involved in complex behaviors and molecular processes underlying neuroplasticity.

Social insects show complex behaviors and master cognitive tasks. The underlying neuronal mechanisms, however, are in most cases only poorly understood due to challenges in monitoring brain activity in freely moving animals. Immediate early genes (IEGs) that get rapidly and transiently expressed following neuronal stimulation provide a powerful tool for detecting behavior-related neuronal activity in vertebrates. In social insects, like honey bees, and in insects in general, this approach is not yet routinely established, even though these genes are highly conserved. First studies revealed a vast potential of using IEGs as neuronal activity markers to analyze the localization, function, and plasticity of neuronal circuits underlying complex social behaviors. We summarize the current knowledge on IEGs in social insects and provide ideas for future research directions.

Distributed plasticity in ant visual pathways following colour learning.

Colour processing at early stages of visual pathways is a topic of intensive study both in vertebrate and invertebrate species. However, it is still unclear how colour learning and memory formation affects an insect brain in the peripheral processing stages and high-order integration centres, and whether associative colour experiences are reflected in plasticity of underlying neuronal circuits. To address this issue, we used Camponotus blandus ants as their proven colour learning and memory capabilities, precisely controllable age and experience, and already known central visual pathways offer unique access to analyse plasticity in neuronal circuits for colour vision in a miniature brain. The potential involvement of distinct neuropils-optic lobes (OLs), mushroom body (MB) input (collar) and output (vertical lobe), anterior optic tubercle (AOTU) and central complex (CX)-in associative colour experiences was assessed by quantification of volumetric and synaptic changes (MB collar) directly after colour conditioning and, 3 days later, after the establishment of long-term memory (LTM). To account for potential effects of non-associative light exposure, we compared neuronal changes in the brain of colour-naive foragers with those of foragers that had been exposed to light in a non-associative way. The results clearly show that the OLs, AOTU, and CX respond with plastic changes after colour learning and LTM formation. This suggests a complex neuronal network for colour learning and memory formation involving multiple brain levels. Such a colour-processing network probably represents an efficient design promoting fast and accurate behavioural decisions during orientation and navigation.

UV light perception is modulated by the odour element of an olfactory-visual compound in restrained honeybees.

Honeybees use visual and olfactory cues to detect flowers during foraging trips. Hence, the reward association of a nectar source is a multimodal construct which has at least two major components - olfactory and visual cues. How both sensory modalities are integrated to form a common reward association and whether and how they may interfere, is an open question. The present study used stimulation with UV, blue and green light to evoke distinct photoreceptor activities in the compound eye and two odour components (geraniol, citronellol). To test if a compound of both modalities is perceived as the sum of its elements (elemental processing) or as a unique cue (configural processing), we combined monochromatic light with single odour components in positive (PP) and negative patterning (NP) experiments. During PP, the compound of two modalities was rewarded, whereas the single elements were not. For NP, stimuli comprising a single modality were rewarded, whereas the olfactory-visual compound was not. Furthermore, we compared the differentiation abilities between two light stimuli that were or were not part of an olfactory-visual compound. Interestingly, the behavioural performances revealed a prominent case of configural processing, but only in those cases when UV light was an element of an olfactory-visual compound. Instead, learning with green- and blue-containing compounds rather supports elemental processing theory.

Early foraging life: spatial and temporal aspects of landmark learning in the ant Cataglyphis noda.

Within the powerful navigational toolkit implemented in desert ants, path integration and landmark guidance are the key routines. Here, we use cue-conflict experiments to investigate the interplay between these two routines in ants, Cataglyphis noda, which start their foraging careers (novices) with learning walks and are then tested at different stages of experience. During their learning walks, the novices take nest-centered views from various directions around the nest. In the present experiments, these learning walks are spatially restricted by arranging differently sized water moats around the nest entrance and thus, limiting the space available around the nest and the nest-feeder route. First, we show that the ants are able to return to the nest by landmark guidance only when the novices have had enough space around the nest entrance for properly performing their learning walks. Second, in 180° cue-conflict situations between path integration and landmark guidance, path integration dominates in the beginning of foraging life (after completion of the learning walks), but with increasing numbers of visits to a familiar feeder landmark guidance comes increasingly into play.

Species-specific differences in the fine structure of learning walk elements in Cataglyphis ants.

Cataglyphis desert ants are famous navigators. Like all central place foragers, they are confronted with the challenge to return home, i.e. relocate an inconspicuous nest entrance in the ground, after their extensive foraging trips. When leaving the underground nest for the first time, desert ants perform a striking behavior, so-called learning walks that are well structured. However, it is still unclear how the ants initially acquire the information needed for sky- and landmark-based navigation, in particular how they calibrate their compass system at the beginning of their foraging careers. Using high-speed video analyses, we show that different Cataglyphis species include different types of characteristic turns in their learning walks. Pirouettes are full or partial rotations (tight turns about the vertical body axis) during which the ants frequently stop and gaze back in the direction of the nest entrance during the longest stopping phases. In contrast, voltes are small walked circles without directed stopping phases. Interestingly, only Cataglyphis ant species living in a cluttered, and therefore visually rich, environment (i.e. C. noda and C. aenescens in southern Greece) perform both voltes and pirouettes. They look back to the nest entrance during pirouettes, most probably to take snapshots of the surroundings. In contrast, C. fortis inhabiting featureless saltpans in Tunisia perform only voltes and do not stop during these turns to gaze back at the nest - even if a set of artificial landmarks surrounds the nest entrance.

Innate colour preference, individual learning and memory retention in the ant Camponotus blandus.

Ants are a well-characterized insect model for the study of visual learning and orientation, but the extent to which colour vision is involved in these tasks remains unknown. We investigated the colour preference, learning and memory retention of Camponotus blandus foragers under controlled laboratory conditions. Our results show that C. blandus foragers exhibit a strong innate preference for ultraviolet (UV, 365 nm) over blue (450 nm) and green (528 nm) wavelengths. The ants can learn to discriminate 365 nm from either 528 nm or 450 nm, independent of intensity changes. However, they fail to discriminate between 450 nm and 528 nm. Modelling of putative colour spaces involving different numbers of photoreceptor types revealed that colour discrimination performance of individual ants is best explained by dichromacy, comprising a short-wavelength (UV) receptor with peak sensitivity at about 360 nm, and a long-wavelength receptor with peak sensitivity between 470 nm and 560 nm. Foragers trained to discriminate blue or green from UV light are able to retain the learned colour information in an early mid-term (e-MTM), late mid-term (l-MTM), early long-term (e-LTM) and late long-term (l-LTM) memory from where it can be retrieved after 1 h, 12 h, 24 h, 3 days and 7 days after training, indicating that colour learning may induce different memory phases in ants. Overall, our results show that ants can use chromatic information in a way that should promote efficient foraging in complex natural environments.

Separation of different pollen types by chemotactile sensing in Bombus terrestris.

When tasting food, animals rely on chemical and tactile cues, which determine the animal's decision on whether to eat food. As food nutritional composition has enormous consequences for the survival of animals, food items should generally be tasted before they are eaten or collected for later consumption. Even though recent studies have confirmed the importance of, for example, gustatory cues, compared with olfaction only little is known about the representation of chemotactile stimuli at the receptor level (let alone higher brain centers) in animals other than vertebrates. To better understand how invertebrates may process chemotactile cues, we used bumblebees as a model species and combined electroantennographical (EAG) recordings with a novel technique for chemotactile antennal stimulation in bees. The recorded EAG responses to chemotactile stimulation clearly separated volatile compounds by both compound identity and concentration, and could be successfully applied to test the receptor activity evoked by different types of pollen. We found that two different pollen types (apple and almond; which were readily distinguished by bumblebees in a classical conditioning task) evoked significantly distinct neural activity already at the antennal receptor level. Our novel stimulation technique therefore enables investigation of chemotactile sensing, which is highly important for assessing food nutritional quality while foraging. It can further be applied to test other chemosensory behaviors, such as mate or nest mate recognition, or to investigate whether toxic substances, e.g. in pollen, affect neuronal separation of different food types.

Ontogeny of learning walks and the acquisition of landmark information in desert ants, Cataglyphis fortis.

At the beginning of their foraging lives, desert ants (Cataglyphis fortis) are for the first time exposed to the visual world within which they henceforth must accomplish their navigational tasks. Their habitat, North African salt pans, is barren, and the nest entrance, a tiny hole in the ground, is almost invisible. Although natural landmarks are scarce and the ants mainly depend on path integration for returning to the starting point, they can also learn and use landmarks successfully to navigate through their largely featureless habitat. Here, we studied how the ants acquire this information at the beginning of their outdoor lives within a nest-surrounding array of three artificial black cylinders. Individually marked 'newcomers' exhibit a characteristic sequence of learning walks. The meandering learning walks covering all directions of the compass first occur only within a few centimeters of the nest entrance, but then increasingly widen, until after three to seven learning walks, foraging starts. When displaced to a distant test field in which an identical array of landmarks has been installed, the ants shift their search density peaks more closely to the fictive goal position, the more learning walks they have performed. These results suggest that learning of a visual landmark panorama around a goal is a gradual rather than an instantaneous process.

Neuropeptidomics of the carpenter ant Camponotus floridanus.

Ants show a rich behavioral repertoire and a highly complex organization, which have been attracting behavioral and sociobiological researchers for a long time. The neuronal underpinnings of ant behavior and social organization are, however, much less understood. Neuropeptides are key signals that orchestrate animal behavior and physiology, and it is thus feasible to assume that they play an important role also for the social constitution of ants. Despite the availability of different ant genomes and in silico prediction of ant neuropeptides, a comprehensive biochemical survey of the neuropeptidergic communication possibilities of ants is missing. We therefore combined different mass spectrometric methods to characterize the neuropeptidome of the adult carpenter ant Camponotus floridanus. We also characterized the local neuropeptide complement in different parts of the nervous and neuroendocrine system, including the antennal and optic lobes. Our analysis identifies 39 neuropeptides encoded by different prepropeptide genes, and in silico predicts new prepropeptide genes encoding CAPA peptides, CNMamide as well as homologues of the honey bee IDLSRFYGHFNT- and ITGQGNRIF-containing peptides. Our data provides basic information about the identity and localization of neuropeptides that is required to anatomically and functionally address the role and significance of neuropeptides in ant behavior and physiology.