Why is it so difficult to study magnetic compass orientation in murine rodents?

A magnetic compass sense has been demonstrated in all major classes of vertebrates, as well as in many invertebrates. In mammals, controlled laboratory studies of mice have provided evidence for a robust magnetic compass that is comparable to, or exceeds, the performance of that in other animals. Nevertheless, the vast majority of laboratory studies of spatial behavior and cognition in murine rodents have failed to produce evidence of sensitivity to magnetic cues. Given the central role that a magnetic compass sense plays in the spatial ecology and cognition of non-mammalian vertebrates, and the potential utility that a global/universal reference frame derived from the magnetic field would have in mammals, the question of why responses to magnetic cues have been so difficult to demonstrate reliably is of considerable importance. In this paper, we review evidence that the magnetic compass of murine rodents shares a number of properties with light-dependent compasses in a wide variety of other animals generally believed to be mediated by a radical pair mechanism (RPM) or related quantum process. Consistent with the RPM, we summarize both published and previously unpublished findings suggesting that the murine rodent compass is sensitive to low-level radio frequency (RF) fields. Finally, we argue that the presence of anthropogenic RF fields in laboratory settings, may be an important source of variability in responses of murine rodents to magnetic cues.

Polarized light modulates light-dependent magnetic compass orientation in birds.

Magnetoreception of the light-dependent magnetic compass in birds is suggested to be mediated by a radical-pair mechanism taking place in the avian retina. Biophysical models on magnetic field effects on radical pairs generally assume that the light activating the magnetoreceptor molecules is nondirectional and unpolarized, and that light absorption is isotropic. However, natural skylight enters the avian retina unidirectionally, through the cornea and the lens, and is often partially polarized. In addition, cryptochromes, the putative magnetoreceptor molecules, absorb light anisotropically, i.e., they preferentially absorb light of a specific direction and polarization, implying that the light-dependent magnetic compass is intrinsically polarization sensitive. To test putative interactions between the avian magnetic compass and polarized light, we developed a spatial orientation assay and trained zebra finches to magnetic and/or overhead polarized light cues in a four-arm "plus" maze. The birds did not use overhead polarized light near the zenith for sky compass orientation. Instead, overhead polarized light modulated light-dependent magnetic compass orientation, i.e., how the birds perceive the magnetic field. Birds were well oriented when tested with the polarized light axis aligned parallel to the magnetic field. When the polarized light axis was aligned perpendicular to the magnetic field, the birds became disoriented. These findings are the first behavioral evidence to our knowledge for a direct interaction between polarized light and the light-dependent magnetic compass in an animal. They reveal a fundamentally new property of the radical pair-based magnetoreceptor with key implications for how birds and other animals perceive the Earth's magnetic field.

A mimicked bacterial infection prolongs stopover duration in songbirds-but more pronounced in short- than long-distance migrants.

Migration usually consists of intermittent travel and stopovers, the latter being crucially important for individuals to recover and refuel to successfully complete migration. Quantifying how sickness behaviours influence stopovers is crucial for our understanding of migration ecology and how diseases spread. However, little is known about infections in songbirds, which constitute the majority of avian migrants. We experimentally immune-challenged autumn migrating passerines (both short- and long-distance migrating species) with a simulated bacterial infection. Using an automated radiotelemetry system in the stopover area, we subsequently quantified stopover duration, "bush-level" activity patterns (0.1-30 m) and landscape movements (30-6,000 m). We show that compared to controls, immune-challenged birds prolonged their stopover duration by on average 1.2 days in long-distance and 2.9 days in short-distance migrants, respectively (100%-126% longer than controls, respectively). During the prolonged stopover, the immune-challenged birds kept a high "bush-level" activity (which was unexpected) but reduced their local movements, independent of migration strategy. Baseline immune function, but not blood parasite infections prior to the immune challenge, had a prolonging effect on stopover duration, particularly in long-distance migrants. We conclude that a mimicked bacterial infection does not cause lethargy, per se, but restricts landscape movements and prolongs stopover duration, and that this behavioural response also depends on the status of baseline immune function and migration strategy. This adds a new level to the understanding of how acute inflammation affect migration behaviour and hence the ecology and evolution of migration. Accounting for these effects of bacterial infections will also enable us to fine-tune and apply optimal migration theory. Finally, it will help us predicting how migrating animals may respond to increased pathogen pressure caused by global change.

Immune function and blood parasite infections impact stopover ecology in passerine birds.

Stopovers play a crucial role for the success of migrating animals and are key to optimal migration theory. Variation in refuelling rates, stopover duration and departure decisions among individuals has been related to several external factors. The physiological mechanisms shaping stopover ecology are, however, less well understood. Here, we explore how immune function and blood parasite infections relate to several aspects of stopover behaviour in autumn migrating short- and long-distance migrating songbirds. We blood sampled individuals of six species and used an automated radio-telemetry system in the stopover area to subsequently quantify stopover duration, 'bush-level' activity patterns (~ 0.1-30 m), landscape movements (~ 30-6000 m), departure direction and departure time. We show that complement activity, the acute phase protein haptoglobin and blood parasite infections were related to prolonged stopover duration. Complement activity (i.e., lysis) and total immunoglobulins were negatively correlated with bush-level activity patterns. The differences partly depended on whether birds were long-distance or short-distance migrants. Birds infected with avian malaria-like parasites showed longer landscape movements during the stopover than uninfected individuals, and birds with double blood parasite infections departed more than 2.5 h later after sunset/sunrise suggesting shorter flight bouts. We conclude that variation in baseline immune function and blood parasite infection status affects stopover ecology and helps explain individual variation in stopover behaviour. These differences affect overall migration speed, and thus can have significant impact on migration success and induce carry-over effects on other annual-cycle stages. Immune function and blood parasites should, therefore, be considered as important factors when applying optimal migration theory.

Zebra finches have a light-dependent magnetic compass similar to migratory birds.

Birds have a light-dependent magnetic compass that provides information about the spatial alignment of the geomagnetic field. It is proposed to be located in the avian retina and mediated by a light-induced, radical-pair mechanism involving cryptochromes as sensory receptor molecules. To investigate how the behavioural responses of birds under different light spectra match with cryptochromes as the primary magnetoreceptor, we examined the spectral properties of the magnetic compass in zebra finches. We trained birds to relocate a food reward in a spatial orientation task using magnetic compass cues. The birds were well oriented along the trained magnetic compass axis when trained and tested under low-irradiance 521 nm green light. In the presence of a 1.4 MHz radio-frequency electromagnetic (RF)-field, the birds were disoriented, which supports the involvement of radical-pair reactions in the primary magnetoreception process. Birds trained and tested under 638 nm red light showed a weak tendency to orient ∼45 deg clockwise of the trained magnetic direction. Under low-irradiance 460 nm blue light, they tended to orient along the trained magnetic compass axis, but were disoriented under higher irradiance light. Zebra finches trained and tested under high-irradiance 430 nm indigo light were well oriented along the trained magnetic compass axis, but disoriented in the presence of a RF-field. We conclude that magnetic compass responses of zebra finches are similar to those observed in nocturnally migrating birds and agree with cryptochromes as the primary magnetoreceptor, suggesting that light-dependent, radical-pair-mediated magnetoreception is a common property for all birds, including non-migratory species.

Pre-settlement coral-reef fish larvae respond to magnetic field changes during the day.

Observations of coral-reef fish larvae have revealed remarkably consistent orientation behaviour while swimming offshore, requiring large-scale orientation cues. However, the mechanisms underlying this behaviour are still being investigated. One potential large-scale cue for orientation is the Earth's geomagnetic field. Here, we examined the effect of magnetic field manipulations on the orientation behaviour of coral-reef fish during the pelagic larval phase. In the absence of visual cues, individual larvae responded to a 90 deg shift of the horizontal component of the magnetic field within a Helmholtz coil with a comparable shift in orientation, demonstrating that they use a magnetic compass for orientation. Our findings suggest that geomagnetic field information guides swimming behaviour of larval fish in the pre-settlement phase. The ability to use large-scale sensory cues allows location-independent orientation of swimming, a behaviour that influences dispersal and connectivity of fish populations, which has important ecological implications for anthropogenic development of marine areas.

No response to linear polarization cues in operant conditioning experiments with zebra finches.

Many animals can use the polarization of light in various behavioural contexts. Birds are well known to use information from the skylight polarization pattern for orientation and compass calibration. However, there are few controlled studies of polarization vision in birds, and the majority of them have not been successful in convincingly demonstrating polarization vision. We used a two-alternative forced choice conditioning approach to assess linear polarization vision in male zebra finches in the 'visible' spectral range (wavelengths >400 nm). The birds were trained to discriminate colour, brightness and polarization stimuli presented on either one of two LCD-screens. All birds were able to discriminate the colour and brightness stimuli, but they were unable to discriminate the polarization stimuli. Our results suggest that in the behavioural context studied here, zebra finches are not able to discriminate polarized light stimuli.

Response of a free-flying songbird to an experimental shift of the light polarization pattern around sunset.

The magnetic field, the sun, the stars and the polarization pattern of visible light during twilight are important cues for orientation in nocturnally migrating songbirds. As these cues change with time and location on Earth, the polarization pattern was put forward as a likely key reference system calibrating the other compass systems. Whether this applies generally to migratory birds is, however, controversially discussed. We used an experimental approach in free-flying birds to study the role of polarization for their departure direction in autumn. Experimental birds experienced a 90 deg shift of the band of maximum polarization during sunset, whereas control birds experienced the polarization pattern as under natural conditions. Full view of the sunset cues near the horizon was provided during the cue conflict exposure. Here we show that both the experimental and the control birds being released after nautical twilight departed consistently towards south-southeast. Radiotelemetry allowed tracking of the first 15 km of the birds' outward journey, thus the intrinsic migration direction as chosen by the birds was measured. We found no recalibration of the magnetic compass after pre-exposure to a cue conflict between the natural magnetic field and the artificially shifted polarization pattern at sunset. The lacking difference in the departure direction of both groups may suggest that birds did not recalibrate any of the compass systems during the experiment. As free-flying migrants can use all available orientation cues after release, it remains unknown whether our birds might have used the magnetic and/or star compass to determine their departure direction.

Spontaneous magnetic orientation in larval Drosophila shares properties with learned magnetic compass responses in adult flies and mice.

We provide evidence for spontaneous quadramodal magnetic orientation in a larval insect. Second instar Berlin, Canton-S and Oregon-R × Canton-S strains of Drosophila melanogaster exhibited quadramodal orientation with clusters of bearings along the four anti-cardinal compass directions (i.e. 45, 135, 225 and 315 deg). In double-blind experiments, Canton-S Drosophila larvae also exhibited quadramodal orientation in the presence of an earth-strength magnetic field, while this response was abolished when the horizontal component of the magnetic field was cancelled, indicating that the quadramodal behavior is dependent on magnetic cues, and that the spontaneous alignment response may reflect properties of the underlying magnetoreception mechanism. In addition, a re-analysis of data from studies of learned magnetic compass orientation by adult Drosophila melanogaster and C57BL/6 mice revealed patterns of response similar to those exhibited by larval flies, suggesting that a common magnetoreception mechanism may underlie these behaviors. Therefore, characterizing the mechanism(s) of magnetoreception in flies may hold the key to understanding the magnetic sense in a wide array of terrestrial organisms.

Effects of low-level RF fields reveal complex pattern of magnetic input to the avian magnetic compass.

The avian magnetic compass can be disrupted by weak narrow-band and broadband radio-frequency (RF) fields in the lower MHz range. However, it is unclear whether disruption of the magnetic compass results from the elimination of the perception pattern produced by the magnetic field or from qualitative changes that make the pattern unrecognizable. We show that zebra finches trained in a 4-arm maze to orient relative to the magnetic field are disoriented when tested in the presence of low-level (~ 10 nT) Larmor-frequency RF fields. However, they are able to orient when tested in such RF fields if trained under this condition, indicating that the RF field alters, but does not eliminate, the magnetic input. Larmor-frequency RF fields of higher intensities, with or without harmonics, dramatically alter the magnetic compass response. In contrast, exposure to broadband RF fields in training, in testing, or in both training and testing eliminates magnetic compass information. These findings demonstrate that low-level RF fields at intensities found in many laboratory and field experiments may have very different effects on the perception of the magnetic field in birds, depending on the type and intensity of the RF field, and the birds' familiarity with the RF-generated pattern.

Migratory navigation in birds: new opportunities in an era of fast-developing tracking technology.

Birds have remained the dominant model for studying the mechanisms of animal navigation for decades, with much of what has been discovered coming from laboratory studies or model systems. The miniaturisation of tracking technology in recent years now promises opportunities for studying navigation during migration itself (migratory navigation) on an unprecedented scale. Even if migration tracking studies are principally being designed for other purposes, we argue that attention to salient environmental variables during the design or analysis of a study may enable a host of navigational questions to be addressed, greatly enriching the field. We explore candidate variables in the form of a series of contrasts (e.g. land vs ocean or night vs day migration), which may vary naturally between migratory species, populations or even within the life span of a migrating individual. We discuss how these contrasts might help address questions of sensory mechanisms, spatiotemporal representational strategies and adaptive variation in navigational ability. We suggest that this comparative approach may help enrich our knowledge about the natural history of migratory navigation in birds.

Cryptochrome expression in avian UV cones: revisiting the role of CRY1 as magnetoreceptor.

Cryptochromes (CRY) have been proposed as putative magnetoreceptors in vertebrates. Localisation of CRY1 in the UV cones in the retinas of birds suggested that it could be the candidate magnetoreceptor. However, recent findings argue against this possibility. CRY1 is a type II cryptochrome, a subtype of cryptochromes that may not be inherently photosensitive, and it exhibits a clear circadian expression in the retinas of birds. Here, we reassessed the localisation and distribution of CRY1 in the retina of the zebra finch. Zebra finches have a light-dependent magnetic compass based on a radical-pair mechanism, similar to migratory birds. We found that CRY1 colocalised with the UV/V opsin (SWS1) in the outer segments of UV cones, but restricted to the tip of the segments. CRY1 was found in all UV cones across the entire retina, with the highest densities near the fovea. Pre-exposure of birds to different wavelengths of light did not result in any difference in CRY1 detection, suggesting that CRY1 did not undergo any detectable functional changes as result of light activation. Considering that CRY1 is likely not involved in magnetoreception, our findings open the possibility for an involvement in different, yet undetermined functions in the avian UV/V cones.

Bird migration: Clock and compass facilitate hemisphere switching.

After cross-equatorial wintering, migratory birds reliably return to their natal grounds, but a population of cliff swallows recently switched breeding hemisphere. They inverted their annual cycle and migration directions almost instantaneously.

A behavioral perspective on the biophysics of the light-dependent magnetic compass: a link between directional and spatial perception?

In terrestrial organisms, sensitivity to the Earth's magnetic field is mediated by at least two different magnetoreception mechanisms, one involving biogenic ferromagnetic crystals (magnetite/maghemite) and the second involving a photo-induced biochemical reaction that forms long-lasting, spin-coordinated, radical pair intermediates. In some vertebrate groups (amphibians and birds), both mechanisms are present; a light-dependent mechanism provides a directional sense or 'compass', and a non-light-dependent mechanism underlies a geographical-position sense or 'map'. Evidence that both magnetite- and radical pair-based mechanisms are present in the same organisms raises a number of interesting questions. Why has natural selection produced magnetic sensors utilizing two distinct biophysical mechanisms? And, in particular, why has natural selection produced a compass mechanism based on a light-dependent radical pair mechanism (RPM) when a magnetite-based receptor is well suited to perform this function? Answers to these questions depend, to a large degree, on how the properties of the RPM, viewed from a neuroethological rather than a biophysical perspective, differ from those of a magnetite-based magnetic compass. The RPM is expected to produce a light-dependent, 3-D pattern of response that is axially symmetrical and, in some groups of animals, may be perceived as a pattern of light intensity and/or color superimposed on the visual surroundings. We suggest that the light-dependent magnetic compass may serve not only as a source of directional information but also provide a spherical coordinate system that helps to interface metrics of distance, direction and spatial position.

Orientation and autumn migration routes of juvenile sharp-tailed sandpipers at a staging site in Alaska.

Arctic waders are well known for their impressive long-distance migrations between their high northerly breeding grounds and wintering areas in the Southern hemisphere. Performing such long migrations requires precise orientation mechanisms. We conducted orientation cage experiments with juvenile sharp-tailed sandpipers (Calidris acuminata) to investigate what cues they rely on when departing from Alaska on their long autumn migration flights across the Pacific Ocean to Australasia, and which possible migration routes they could use. Experiments were performed under natural clear skies, total overcast conditions and in manipulated magnetic fields at a staging site in Alaska. Under clear skies the juvenile sharp-tailed sandpipers oriented towards SSE, which coincides well with reported sun compass directions from their breeding grounds in Siberia towards Alaska and could reflect their true migratory direction towards Australasia assuming that they change direction towards SW somewhere along the route. Under overcast skies the sandpipers showed a mean direction towards SW which would lead them to Australasia, if they followed a sun compass route. However, because of unfavourable weather conditions (headwinds) associated with overcast conditions, these south-westerly directions could also reflect local movements. The juvenile sharp-tailed sandpipers responded clearly to the manipulated magnetic field under overcast skies, suggesting the use of a magnetic compass for selecting their courses.

White-throated sparrows calibrate their magnetic compass by polarized light cues during both autumn and spring migration.

The interaction and hierarchy of celestial and magnetic compass cues used by migratory songbirds for orientation has long been the topic of an intense debate. We have previously shown that migratory Savannah sparrows, Passerculus sandwichensis, use polarized light cues near the horizon at sunrise and sunset to recalibrate their magnetic compass. Birds exposed to a +/-90 deg. shifted artificial polarization pattern at sunrise or sunset recalibrated their magnetic compass, but only when given full access to celestial cues, including polarized light cues near the horizon. In the current study, we carried out cue conflict experiments with white-throated sparrows, Zonotrichia albicollis, during both spring and autumn migration in a transition zone between the species' breeding and wintering areas on the south shore of Lake Ontario. We show that white-throated sparrows also recalibrate their magnetic compass by polarized light cues at sunrise and sunset. Sunrise exposure to an artificial polarization pattern shifted relative to the natural magnetic field or exposure to a shift of the magnetic field relative to the natural sky both led to recalibration of the magnetic compass, demonstrating that artificial polarizing filters do not create an anomalous, unnatural orientation response. Our results further indicate that there is no evidence for a difference in compass hierarchy between different phases of migration, confirming previous work showing that polarized light cues near the horizon at sunrise and sunset provide the primary calibration reference both in the beginning and at the end of migration.

Ectosymbionts alter spontaneous responses to the Earth's magnetic field in a crustacean.

Magnetic sensing is used to structure every-day, non-migratory behaviours in many animals. We show that crayfish exhibit robust spontaneous magnetic alignment responses. These magnetic behaviours are altered by interactions with Branchiobdellidan worms, which are obligate ectosymbionts. Branchiobdellidan worms have previously been shown to have positive effects on host growth when present at moderate densities, and negative effects at relatively high densities. Here we show that crayfish with moderate densities of symbionts aligned bimodally along the magnetic northeast-southwest axis, similar to passive magnetic alignment responses observed across a range of stationary vertebrates. In contrast, crayfish with high symbiont densities failed to exhibit consistent alignment relative to the magnetic field. Crayfish without symbionts shifted exhibited quadramodal magnetic alignment and were more active. These behavioural changes suggest a change in the organization of spatial behaviour with increasing ectosymbiont densities. We propose that the increased activity and a switch to quadramodal magnetic alignment may be associated with the use of systematic search strategies. Such a strategy could increase contact-rates with conspecifics in order to replenish the beneficial ectosymbionts that only disperse between hosts during direct contact. Our results demonstrate that crayfish perceive and respond to magnetic fields, and that symbionts influence magnetically structured spatial behaviour of their hosts.

Dramatic orientation shift of white-crowned sparrows displaced across longitudes in the high Arctic.

Advanced spatial-learning adaptations have been shown for migratory songbirds, but it is not well known how the simple genetic program encoding migratory distance and direction in young birds translates to a navigation mechanism used by adults. A number of convenient cues are available to define latitude on the basis of geomagnetic and celestial information, but very few are useful to defining longitude. To investigate the effects of displacements across longitudes on orientation, we recorded orientation of adult and juvenile migratory white-crowned sparrows, Zonotrichia leucophrys gambelii, after passive longitudinal displacements, by ship, of 266-2862 km across high-arctic North America. After eastward displacement to the magnetic North Pole and then across the 0 degrees declination line, adults and juveniles abruptly shifted their orientation from the migratory direction to a direction that would lead back to the breeding area or to the normal migratory route, suggesting that the birds began compensating for the displacement by using geomagnetic cues alone or together with solar cues. In contrast to predictions by a simple genetic migration program, our experiments suggest that both adults and juveniles possess a navigation system based on a combination of celestial and geomagnetic information, possibly declination, to correct for eastward longitudinal displacements.

Expression patterns of cryptochrome genes in avian retina suggest involvement of Cry4 in light-dependent magnetoreception.

The light-dependent magnetic compass of birds provides orientation information about the spatial alignment of the geomagnetic field. It is proposed to be located in the avian retina, and be mediated by a light-induced, biochemical radical-pair mechanism involving cryptochromes as putative receptor molecules. At the same time, cryptochromes are known for their role in the negative feedback loop in the circadian clock. We measured gene expression of Cry1, Cry2 and Cry4 in the retina, muscle and brain of zebra finches over the circadian day to assess whether they showed any circadian rhythmicity. We hypothesized that retinal cryptochromes involved in magnetoreception should be expressed at a constant level over the circadian day, because birds use a light-dependent magnetic compass for orientation not only during migration, but also for spatial orientation tasks in their daily life. Cryptochromes serving in circadian tasks, on the other hand, are expected to be expressed in a rhythmic (circadian) pattern. Cry1 and Cry2 displayed a daily variation in the retina as expected for circadian clock genes, while Cry4 expressed at constant levels over time. We conclude that Cry4 is the most likely candidate magnetoreceptor of the light-dependent magnetic compass in birds.

Feasibility of sun and magnetic compass mechanisms in avian long-distance migration.

Birds use different compass mechanisms based on celestial (stars, sun, skylight polarization pattern) and geomagnetic cues for orientation. Yet, much remains to be understood how birds actually use these compass mechanisms on their long-distance migratory journeys. Here, we assess in more detail the consequences of using different sun and magnetic compass mechanisms for the resulting bird migration routes during both autumn and spring migration. First, we calculated predicted flight routes to determine which of the compasses mechanisms lead to realistic and feasible migration routes starting at different latitudes during autumn and spring migration. We then compared the adaptive values of the different compass mechanisms by calculating distance ratios in relation to the shortest possible trajectory for three populations of nocturnal passerine migrants: northern wheatear Oenanthe oenanthe, pied flycatcher Ficedula hypoleuca, and willow warbler Phylloscopus trochilus. Finally, we compared the predicted trajectories for different compass strategies with observed routes based on recent light-level geolocation tracking results for five individuals of northern wheatears migrating between Alaska and tropical Africa. We conclude that the feasibility of different compass routes varies greatly with latitude, migratory direction, migration season, and geographic location. Routes following a single compass course throughout the migratory journey are feasible for many bird populations, but the underlying compass mechanisms likely differ between populations. In many cases, however, the birds likely have to reorient once to a few times along the migration route and/or use map information to successfully reach their migratory destination.