techniques-for-investigating-the-anatomy-of-the-ant-visual-system

Techniques for Investigating the Anatomy of the Ant Visual System

This article outlines a suite of techniques in light microscopy (LM) and electron microscopy (EM) which can be used to study the internal and external eye ...

Macquarie University

Estimation of distance travelled (odometry) forms a vital part of navigation for solitarily foraging ants. In this study we investigated the properties of odometric memory in the Australian desert ant Melophorus bagoti. Ants were trained to travel in linear channels to a feeder placed at 6 m or 12 m from the nest. We determined if the ability to estimate distances accurately increased with experience. We also determined the delay at which the odometric memory started to decay at both these distances. Ants with six trials of experience did not get better at estimating distances and the odometric memory decayed after a 24 h delay. We then determined if ants integrated their latest odometric memory with their previous memories. We did this by training two groups of ants for five trials, one group to 6 m and another to 12 m, and then halving or doubling the outbound distance on their sixth trip, respectively. The ant's estimate was noted when the ants were released either immediately or after a 24 h delay. Ants always estimated their last outbound distance when released immediately. However, they switched to route-based navigation rather than estimating distances after a 24 h delay (at which time their odometric memory would have decayed).

Acquiring, retaining and integrating memories of the outbound distance in the Australian desert ant Melophorus bagoti.

Highly evolved eusocial insects such as ants return from a food source to their nest by the shortest possible distance. This form of navigation, called path-integration, involves keeping track of the distance travelled and the angles steered on the outbound journey, which then aids in the computation of the shortest return distance. In featureless terrain, ants rely on the path integrator to travel the entire distance to return to the nest, whereas in landmark-rich habitats ants are guided by visual cues and in the absence of the visual cues homing ants rely on the path integrator to travel only an initial 10-60 cm of the homebound distance. The functioning of the path integrator in a habitat of intermediate landmark density is unknown. The findings reported here show that when the outward journey is on a familiar foraging area, and the inward journey is on an unfamiliar area, the Australian route-following desert ant Melophorus bagoti relies on the path integrator and consistently travels half the distance of the outward trip. However, when both the outward and inward trips are performed in plain and featureless channels, which blocks the distinct terrestrial visual cues, ants travel the entire distance accurately. A similar half-way abbreviation of the home vector occurs when the ant's outward trip is in an L-shaped channel and the homeward trip is over an open and unfamiliar region. The ecological significance of these new findings is discussed.

Homing strategies of the Australian desert ant Melophorus bagoti. I. Proportional path-integration takes the ant half-way home.

Individually foraging ants are known to return to their nest by using path-integration and recording visual information present in the environment. The interaction between the path integrator and the information provided by the visual cues in an Australian desert ant are reported here. Ants were trained to travel in a 1-m wide and 20-m long corridor of cylinders. Homeward paths of trained ants were recorded in the presence and absence of vector information and route cues in both the familiar training field and in an unfamiliar test field. Homing ants used route cue information only in a familiar context. The route cues were not essential but served to reduce the deviation of the homing trajectory from the nest-feeder line. When displaced locally, homebound ants initially oriented towards the nest using distant cues and then headed in a direction intermediate between that dictated by the path integrator and the distant cues. If in the course of travel ants encountered the familiar path they adhered to it. If not, they travelled on average half the distance of the outbound journey and initiated a search directed towards the nest. Following the search, ants headed in a direction intermediate between that dictated by the route cues and the distant cues. In an unfamiliar context neither vector nor route cue information could steer a homing ant towards the nest. The dominance of distant cues, the importance of familiar context and the interaction between different navigation strategies are discussed here.

Homing strategies of the Australian desert ant Melophorus bagoti. II. Interaction of the path integrator with visual cue information.

Search is an important tool in an ant's navigational toolbox to relocate food sources and find the inconspicuous nest entrance. In habitats where landmark information is sparse, homing ants travel their entire home vector before searching systematically with ever increasing loops. Search strategies have not been previously investigated in ants that inhabit landmark-rich habitats where they typically establish stereotypical routes. Here we examine the search strategy in one such ant, Melophorus bagoti, by confining their foraging in one-dimensional channels to determine if their search pattern changes with experience, location of distant cues and altered distance on the homebound journey. Irrespective of conditions, we found ants exhibit a progressive search that drifted towards the fictive nest and beyond. Segments moving away from the start of the homeward journey were longer than segments heading back towards the start. The right tail distribution of segment lengths was well fitted by a power function, but slopes less than -3 on a log-log plot indicate that the process cannot be characterized as Lévy searches that have optimal slopes near -2. A double exponential function fits the distribution of segment lengths better, supporting another theoretically optimal search pattern, the composite Brownian walk.

Search strategies of ants in landmark-rich habitats.

The Central Australian desert ant Melophorus bagoti is the most thermophilic ant on the continent. It comes out to forage during the hottest part of the day in the summer months. The ant shares a cluttered, plant-filled habitat with other arthropods and uses a range of navigational strategies. We review recent studies on this species concerning its use of habitual routes, distant landmarks, landmarks around the nest, and path integration, which is keeping track of the distance and direction traveled from one's starting point. Functional predictions concerning the acquisition, retention, and integration of memories of distances and of landmarks are also reviewed, illuminating the behavioral ecology of spatial cognition.

Traveling in clutter: navigation in the Central Australian desert ant Melophorus bagoti.

The Australian desert ant Melophorus bagoti shows remarkable visual navigational skills relying on visual rather than on chemical cues during their foraging trips. M. bagoti ants travel individually through a visually cluttered environment guided by landmarks as well as by path integration. An examination of their visual system is hence of special interest and we address this here. Workers exhibit distinct size polymorphism and their eye and ocelli size increases with head size. The ants possess typical apposition eyes with about 420-590 ommatidia per eye, a horizontal visual field of approximately 150° and facet lens diameters between 8 and 19 μm, depending on body size, with frontal facets being largest. The average interommatidial angle Δϕ is 3.7°, the average acceptance angle of the rhabdom Δρ(rh) is 2.9°, with average rhabdom diameter of 1.6 μm and the average lens blur at half-width Δρ(l) is 2.3°. With a Δρ(rh)/Δϕ ratio of much less than 2, the eyes undersample the visual scene but provide high contrast, and surprising detail of the landmark panorama that has been shown to be used for navigation.

The properties of the visual system in the Australian desert ant Melophorus bagoti.

Ants are thought to be special among Hymenopterans in having only dichromatic colour vision based on two spectrally distinct photoreceptors. Many ants are highly visual animals, however, and use vision extensively for navigation. We show here that two congeneric day- and night-active Australian ants have three spectrally distinct photoreceptor types, potentially supporting trichromatic colour vision. Electroretinogram recordings show the presence of three spectral sensitivities with peaks (λmax) at 370, 450 and 550 nm in the night-active Myrmecia vindex and peaks at 370, 470 and 510 nm in the day-active Myrmecia croslandi. Intracellular electrophysiology on individual photoreceptors confirmed that the night-active M. vindex has three spectral sensitivities with peaks (λmax) at 370, 430 and 550 nm. A large number of the intracellular recordings in the night-active M. vindex show unusually broad-band spectral sensitivities, suggesting that photoreceptors may be coupled. Spectral measurements at different temporal frequencies revealed that the ultraviolet receptors are comparatively slow. We discuss the adaptive significance and the probability of trichromacy in Myrmecia ants in the context of dim light vision and visual navigation.

Three spectrally distinct photoreceptors in diurnal and nocturnal Australian ants.

In addition to foraging individually several species of ants guide nestmates to a goal by tandem running. We found that the Australian ant, Camponotus consobrinus, forages both individually and by tandem running to head to the same goal, nest-specific native Australian trees on which they forage. While paths of solitary foragers and initial paths of tandem followers showed no differences in heading directions or straightness, tandem followers moved at about half the speed of solitary runs. When leaders were experimentally removed, follower ants initially engaged in a systematic search around the point of interruption, following which they either (a) headed directly towards and successfully reached the foraging trees, or (b) continued searching or (c) returned to the nest. The high incidence of followers that successfully navigated towards the foraging trees on their own provides strong evidence that many tandem followers are in fact experienced foragers. Detailed analysis of the searching behaviour revealed that even seemingly lost followers displayed a directional bias towards the foraging trees in their search path. Our results show that in a foraging context follower ants in a tandem pair are not always naïve.

Follower ants in a tandem pair are not always naïve.

Ants are unusual among insects in that individuals of the same species within a single colony have different modes of locomotion and tasks. We know from walking ants that vision plays a significant role in guiding this behaviour, but we know surprisingly little about the potential contribution of visual sensory structures for a flying mode of locomotion. Here we investigate the structure of the compound eye and ocelli in pedestrian workers, alate females and alate males of an Australian ant, Camponotus consobrinus, and discuss the trade-offs involved in optical sensitivity and spatial resolution. Male ants have more but smaller ommatidia and the smallest interommatidial angles, which is most likely an adaptation to visually track individual flying females. Both walking and flying forms of ants have a similar proportion of specialized receptors sensitive to polarized skylight, but the absolute number of these receptors varies, being greatest in males. Ocelli are present only in the flying forms. Each ocellus consists of a bipartite retina with a horizon-facing dorsal retina, which contains retinula cells with long rhabdoms, and a sky-facing ventral retina with shorter rhabdoms. We discuss the implications of these and their potential for sensing the pattern of polarized skylight.

Compound eye and ocellar structure for walking and flying modes of locomotion in the Australian ant, Camponotus consobrinus.

Ant foragers are known to memorise visual scenes that allow them to repeatedly travel along idiosyncratic routes and to return to specific places. Guidance is provided by a comparison between visual memories and current views, which critically depends on how well the attitude of the visual system is controlled. Here we show that nocturnal bull ants stabilise their head to varying degrees against locomotion-induced body roll movements, and this ability decreases as light levels fall. There are always un-compensated head roll oscillations that match the frequency of the stride cycle. Head roll stabilisation involves both visual and non-visual cues as ants compensate for body roll in complete darkness and also respond with head roll movements when confronted with visual pattern oscillations. We show that imperfect head roll control degrades navigation-relevant visual information and discuss ways in which navigating ants may deal with this problem.

Head roll stabilisation in the nocturnal bull ant Myrmecia pyriformis: implications for visual navigation.

Ants of the Australian genus Myrmecia partition their foraging niche temporally, allowing them to be sympatric with overlapping foraging requirements. We used histological techniques to study the light and dark adaptation mechanisms in the compound eyes of diurnal (Myrmecia croslandi), crepuscular (M. tarsata, M. nigriceps) and nocturnal ants (M. pyriformis). We found that, except in the day-active species, all ants have a variable primary pigment cell pupil that constricts the crystalline cone in bright light to control for light flux. We show for the nocturnal M. pyriformis that the constriction of the crystalline cone by the primary pigment cells is light dependent whereas the opening of the aperture is regulated by an endogenous rhythm. In addition, in the light-adapted eyes of all species, the retinular cell pigment granules radially migrate towards the rhabdom, a process that in both the day-active M. croslandi and the night-active M. pyriformis is driven by ambient light intensity. Visual system properties thus do not restrict crepuscular and night-active ants to their temporal foraging niche, while day-active ants require high light intensities to operate. We discuss the ecological significance of these adaptation mechanisms and their role in temporal niche partitioning.

Light and dark adaptation mechanisms in the compound eyes of Myrmecia ants that occupy discrete temporal niches.

Ants use both terrestrial landmarks and celestial cues to navigate to and from their nest location. These cues persist even as light levels drop during the twilight/night. Here, we determined the compass cues used by a nocturnal bull ant, Myrmecia midas, in which the majority of individuals begin foraging during the evening twilight period. Myrmecia midas foragers with vectors of ≤5 m when displaced to unfamiliar locations did not follow the home vector, but instead showed random heading directions. Foragers with larger home vectors (≥10 m) oriented towards the fictive nest, indicating a possible increase in cue strength with vector length. When the ants were displaced locally to create a conflict between the home direction indicated by the path integrator and terrestrial landmarks, foragers oriented using landmark information exclusively and ignored any accumulated home vector regardless of vector length. When the visual landmarks at the local displacement site were blocked, foragers were unable to orient to the nest direction and their heading directions were randomly distributed. Myrmecia midas ants typically nest at the base of the tree and some individuals forage on the same tree. Foragers collected on the nest tree during evening twilight were unable to orient towards the nest after small lateral displacements away from the nest. This suggests the possibility of high tree fidelity and an inability to extrapolate landmark compass cues from information collected on the tree and at the nest site to close displacement sites.

Compass cues used by a nocturnal bull ant, Myrmecia midas.

The ability of ants to navigate when the visual landmark information is altered has often been tested by creating large and artificial discrepancies in their visual environment. Here, we had an opportunity to slightly modify the natural visual environment around the nest of the nocturnal bull ant Myrmecia pyriformis We achieved this by felling three dead trees, two located along the typical route followed by the foragers of that particular nest and one in a direction perpendicular to their foraging direction. An image difference analysis showed that the change in the overall panorama following the removal of these trees was relatively little. We filmed the behaviour of ants close to the nest and tracked their entire paths, both before and after the trees were removed. We found that immediately after the trees were removed, ants walked slower and were less directed. Their foraging success decreased and they looked around more, including turning back to look towards the nest. We document how their behaviour changed over subsequent nights and discuss how the ants may detect and respond to a modified visual environment in the evening twilight period.This article is part of the themed issue 'Vision in dim light'.

Subtle changes in the landmark panorama disrupt visual navigation in a nocturnal bull ant.

Individual differences in response thresholds to task-related stimuli may be one mechanism driving task allocation among social insect workers. These differences may arise at various stages in the nervous system. We investigate variability in the peripheral nervous system as a simple mechanism that can introduce inter-individual differences in sensory information. In this study we describe size-dependent variation of the compound eyes and the antennae in the ant Temnothorax rugatulus. Head width in T. rugatulus varies between 0.4 and 0.7 mm (2.6-3.8 mm body length). But despite this limited range of worker sizes we find sensory array variability. We find that the number of ommatidia and of some, but not all, antennal sensilla types vary with head width. The antennal array of T. rugatulus displays the full complement of sensillum types observed in other species of ants, although at much lower quantities than other, larger, studied species. In addition, we describe what we believe to be a new type of sensillum in hymenoptera that occurs on the antennae and on all body segments. T. rugatulus has apposition compound eyes with 45-76 facets per eye, depending on head width, with average lens diameters of 16.5 μm, rhabdom diameters of 5.7 μm and inter-ommatidial angles of 16.8°. The optical system of T. rugatulus ommatidia is severely under focussed, but the absolute sensitivity of the eyes is unusually high. We discuss the functional significance of these findings and the extent to which the variability of sensory arrays may correlate with task allocation.

The sensory arrays of the ant, Temnothorax rugatulus.

Solitary foraging ants have a navigational toolkit, which includes the use of both terrestrial and celestial visual cues, allowing individuals to successfully pilot between food sources and their nest. One such celestial cue is the polarization pattern in the overhead sky. Here, we explore the use of polarized light during outbound and inbound journeys and with different home vectors in the nocturnal bull ant, Myrmecia midas. We tested foragers on both portions of the foraging trip by rotating the overhead polarization pattern by ±45°. Both outbound and inbound foragers responded to the polarized light change, but the extent to which they responded to the rotation varied. Outbound ants, both close to and further from the nest, compensated for the change in the overhead e-vector by about half of the manipulation, suggesting that outbound ants choose a compromise heading between the celestial and terrestrial compass cues. However, ants returning home compensated for the change in the e-vector by about half of the manipulation when the remaining home vector was short (1-2 m) and by more than half of the manipulation when the remaining vector was long (more than 4 m). We report these findings and discuss why weighting on polarization cues change in different contexts.

Polarized light use in the nocturnal bull ant, Myrmecia midas.

Visual navigation is a benchmark information processing task that can be used to identify the consequence of being active in dim-light environments. Visual navigational information that animals use during the day includes celestial cues such as the sun or the pattern of polarized skylight and terrestrial cues such as the entire panorama, canopy pattern, or significant salient features in the landscape. At night, some of these navigational cues are either unavailable or are significantly dimmer or less conspicuous than during the day. Even under these circumstances, animals navigate between locations of importance. Ants are a tractable system for studying navigation during day and night because the fine scale movement of individual animals can be recorded in high spatial and temporal detail. Ant species range from being strictly diurnal, crepuscular, and nocturnal. In addition, a number of species have the ability to change from a day- to a night-active lifestyle owing to environmental demands. Ants also offer an opportunity to identify the evolution of sensory structures for discrete temporal niches not only between species but also within a single species. Their unique caste system with an exclusive pedestrian mode of locomotion in workers and an exclusive life on the wing in males allows us to disentangle sensory adaptations that cater for different lifestyles. In this article, we review the visual navigational abilities of nocturnal ants and identify the optical and physiological adaptations they have evolved for being efficient visual navigators in dim-light.

Article	Year
Acquiring, retaining and integrating memories of the outbound distance in the Australian desert ant Melophorus bagoti. The Journal of experimental biology\| PubMed ID: 17267642	2007
Homing strategies of the Australian desert ant Melophorus bagoti. I. Proportional path-integration takes the ant half-way home. The Journal of experimental biology\| PubMed ID: 17488943	2007
Homing strategies of the Australian desert ant Melophorus bagoti. II. Interaction of the path integrator with visual cue information. The Journal of experimental biology\| PubMed ID: 17488944	2007
Search strategies of ants in landmark-rich habitats. Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology\| PubMed ID: 18781312	2008
Traveling in clutter: navigation in the Central Australian desert ant Melophorus bagoti. Behavioural processes\| PubMed ID: 19049857	2009
The properties of the visual system in the Australian desert ant Melophorus bagoti. Arthropod structure & development\| PubMed ID: 21044895	2011
Three spectrally distinct photoreceptors in diurnal and nocturnal Australian ants. Proceedings. Biological sciences\| PubMed ID: 25994678	2015
Follower ants in a tandem pair are not always naïve. Scientific reports\| PubMed ID: 26021611	2015
Compound eye and ocellar structure for walking and flying modes of locomotion in the Australian ant, Camponotus consobrinus. Scientific reports\| PubMed ID: 26975481	2016
Head roll stabilisation in the nocturnal bull ant Myrmecia pyriformis: implications for visual navigation. The Journal of experimental biology\| PubMed ID: 26994172	2016
Light and dark adaptation mechanisms in the compound eyes of Myrmecia ants that occupy discrete temporal niches. The Journal of experimental biology\| PubMed ID: 27535985	2016
Compass cues used by a nocturnal bull ant, Myrmecia midas. The Journal of experimental biology\| PubMed ID: 28183865	2017
Subtle changes in the landmark panorama disrupt visual navigation in a nocturnal bull ant. Philosophical transactions of the Royal Society of London. Series B, Biological sciences\| PubMed ID: 28193813	2017
The sensory arrays of the ant, Temnothorax rugatulus. Arthropod structure & development\| PubMed ID: 28347859	2017
Polarized light use in the nocturnal bull ant, Myrmecia midas. Royal Society open science\| PubMed ID: 28879002	2017
Moving in Dim Light: Behavioral and Visual Adaptations in Nocturnal Ants. Integrative and comparative biology\| PubMed ID: 28985392	2017

Ajay Narendra

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