Adaptive evolution and loss of a putative magnetoreceptor in passerines.

Migratory birds possess remarkable accuracy in orientation and navigation, which involves various compass systems including the magnetic compass. Identifying the primary magnetosensor remains a fundamental open question. Cryptochromes (Cry) have been shown to be magnetically sensitive, and Cry4a from a migratory songbird seems to show enhanced magnetic sensitivity in vitro compared to Cry4a from resident species. We investigate Cry and their potential involvement in magnetoreception in a phylogenetic framework, integrating molecular evolutionary analyses with protein dynamics modelling. Our analysis is based on 363 bird genomes and identifies different selection regimes in passerines. We show that Cry4a is characterized by strong positive selection and high variability, typical characteristics of sensor proteins. We identify key sites that are likely to have facilitated the evolution of an optimized sensory protein for night-time orientation in songbirds. Additionally, we show that Cry4 was lost in hummingbirds, parrots and Tyranni (Suboscines), and thus identified a gene deletion, which might facilitate testing the function of Cry4a in birds. In contrast, the other avian Cry (Cry1 and Cry2) were highly conserved across all species, indicating basal, non-sensory functions. Our results support a specialization or functional differentiation of Cry4 in songbirds which could be magnetosensation.

Double cones in the avian retina form an oriented mosaic which might facilitate magnetoreception and/or polarized light sensing.

To navigate between breeding and wintering grounds, night-migratory songbirds are aided by a light-dependent magnetic compass sense and maybe also by polarized light vision. Although the underlying mechanisms for magnetoreception and polarized light sensing remain unclear, double cone photoreceptors in the avian retina have been suggested to represent the primary sensory cells. To use these senses, birds must be able to separate the directional information from the Earth's magnetic field and/or light polarization from variations in light intensity. Theoretical considerations suggest that this could be best achieved if neighbouring double cones were oriented in an ordered pattern. Therefore, we investigate the orientation patterns of double cones in European robins (Erithacus rubecula) and domestic chickens (Gallus gallus domesticus). We used whole-mounted retinas labelled with double cone markers to quantify the orientations of individual double cones in relation to their nearest neighbours. In both species, our data show that the double cone array is highly ordered: the angles between neighbouring double cones were more likely to be 90°/-90° in the central retina and 180°/0° in the peripheral retina, respectively. The observed regularity in double cone orientation could aid the cells' putative function in light-dependent magnetoreception and/or polarized light sensing.

Broadband 75-85 MHz radiofrequency fields disrupt magnetic compass orientation in night-migratory songbirds consistent with a flavin-based radical pair magnetoreceptor.

The light-dependent magnetic compass sense of night-migratory songbirds can be disrupted by weak radiofrequency fields. This finding supports a quantum mechanical, radical-pair-based mechanism of magnetoreception as observed for isolated cryptochrome 4, a protein found in birds' retinas. The exact identity of the magnetically sensitive radicals in cryptochrome is uncertain in vivo, but their formation seems to require a bound flavin adenine dinucleotide chromophore and a chain of four tryptophan residues within the protein. Resulting from the hyperfine interactions of nuclear spins with the unpaired electrons, the sensitivity of the radicals to radiofrequency magnetic fields depends strongly on the number of magnetic nuclei (hydrogen and nitrogen atoms) they contain. Quantum-chemical calculations suggested that electromagnetic noise in the frequency range 75-85 MHz could give information about the identity of the radicals involved. Here, we show that broadband 75-85 MHz radiofrequency fields prevent a night-migratory songbird from using its magnetic compass in behavioural experiments. These results indicate that at least one of the components of the radical pair involved in the sensory process of avian magnetoreception must contain a substantial number of strong hyperfine interactions as would be the case if a flavin-tryptophan radical pair were the magnetic sensor.

Electromagnetic 0.1-100 kHz noise does not disrupt orientation in a night-migrating songbird implying a spin coherence lifetime of less than 10 µs.

According to the currently prevailing theory, the magnetic compass sense in night-migrating birds relies on a light-dependent radical-pair-based mechanism. It has been shown that radio waves at megahertz frequencies disrupt magnetic orientation in migratory birds, providing evidence for a quantum-mechanical origin of the magnetic compass. Still, many crucial properties, e.g. the lifetime of the proposed magnetically sensitive radical pair, remain unknown. The current study aims to estimate the spin coherence time of the radical pair, based on the behavioural responses of migratory birds to broadband electromagnetic fields covering the frequency band 0.1-100 kHz. A finding that the birds were unable to use their magnetic compass under these conditions would imply surprisingly long-lived (greater than 10 µs) spin coherence. However, we observed no effect of 0.1-100 kHz radiofrequency (RF) fields on the orientation of night-migratory Eurasian blackcaps (Sylvia atricapilla). This suggests that the lifetime of the spin coherence involved in magnetoreception is shorter than the period of the highest frequency RF fields used in this experiment (i.e. approx. 10 µs). This result, in combination with an earlier study showing that 20-450 kHz electromagnetic fields disrupt magnetic compass orientation, suggests that the spin coherence lifetime of the magnetically sensitive radical pair is in the range 2-10 µs.

Very weak oscillating magnetic field disrupts the magnetic compass of songbird migrants.

Previously, it has been shown that long-distance migrants, garden warblers (Sylvia borin), were disoriented in the presence of narrow-band oscillating magnetic field (1.403 MHz OMF, 190 nT) during autumn migration. This agrees with the data of previous experiments with European robins (Erithacus rubecula). In this study, we report the results of experiments with garden warblers tested under a 1.403 MHz OMF with various amplitudes (∼0.4, 1, ∼2.4, 7 and 20 nT). We found that the ability of garden warblers to orient in round arenas using the magnetic compass could be disrupted by a very weak oscillating field, such as an approximate 2.4, 7 and 20 nT OMF, but not by an OMF with an approximate 0.4 nT amplitude. The results of the present study indicate that the sensitivity threshold of the magnetic compass to the OMF lies around 2-3 nT, while in experiments with European robins the birds were disoriented in a 15 nT OMF but could choose the appropriate migratory direction when a 5 nT OMF was added to the stationary magnetic field. The radical-pair model, one of the mainstream theories of avian magnetoreception, cannot explain the sensitivity to such a low-intensity OMF, and therefore, it needs further refinement.