Central neural coding of sky polarization in insects

Many animals rely on a sun compass for spatial orientation and long-range navigation. In addition to the Sun, insects also exploit the polarization pattern and chromatic gradient of the sky for estimating navigational directions. Analysis of polarization-vision pathways in locusts and crickets has shed first light on brain areas involved in sky compass orientation. Detection of sky polarization relies on specialized photoreceptor cells in a small dorsal rim area of the compound eye. Brain areas involved in polarization processing include parts of the lamina, medulla and lobula of the optic lobe and, in the central brain, the anterior optic tubercle, the lateral accessory lobe and the central complex. In the optic lobe, polarization sensitivity and contrast are enhanced through convergence and opponency. In the anterior optic tubercle, polarized-light signals are integrated with information on the chromatic contrast of the sky. Tubercle neurons combine responses to the UV/green contrast and e-vector orientation of the sky and compensate for diurnal changes of the celestial polarization pattern associated with changes in solar elevation. In the central complex, a topographic representation of e-vector tunings underlies the columnar organization and suggests that this brain area serves as an internal compass coding for spatial directions.     

Evidence for the possible existence of a second polarization-vision pathway in the locust brain

For spatial orientation and navigation, many insects derive compass information from the polarization pattern of the blue sky. The desert locust Schistocerca gregaria detects polarized light with a specialized dorsal rim area of its compound eye. In the locust brain, polarized-light signals are passed through the anterior optic tract and tubercle to the central complex which most likely serves as an internal sky compass. Here, we suggest that neurons of a second visual pathway, via the accessory medulla and posterior optic tubercle, also provide polarization information to the central complex. Intracellular recordings show that two types of neuron in this posterior pathway are sensitive to polarized light. One cell type connects the dorsal rim area of the medulla with the medulla and accessory medulla, and a second type connects the bilaterally paired posterior optic tubercles. Given the evidence for a role of the accessory medulla as the master clock controlling circadian changes in behavioral activity in flies and cockroaches, our data open the possibility that time-compensated polarized-light signals may reach the central complex via this pathway for time-compensated sky-compass navigation.     

Surgical lesion of the anterior optic tract abolishes polarotaxis in tethered flying locusts, <Emphasis Type="Italic">Schistocerca gregaria</Emphasis>

Many insects can detect the polarization pattern of the blue sky and rely on polarization vision for sky compass orientation. In laboratory experiments, tethered flying locusts perform periodic changes in flight behavior under a slowly rotating polarizer even if one eye is painted black. Anatomical tracing studies and intracellular recordings have suggested that the polarization vision pathway in the locust brain involves the anterior optic tract and tubercle, the lateral accessory lobe, and the central complex of the brain. To investigate whether visual pathways through the anterior optic tract mediate polarotaxis in the desert locust, we transected the tract on one side and tested polarotaxis (1) with both eyes unoccluded and (2) with the eye of the intact hemisphere painted black. In the second group of animals, but not in the first group, polarotaxis was abolished. Sham operations did not impair polarotaxis. The experiments show that the anterior optic tract is an indispensable part of visual pathways mediating polarotaxis in the desert locust.     

A distinct layer of the medulla integrates sky compass signals in the brain of an insect

Mass migration of desert locusts is a common phenomenon in North Africa and the Middle East but how these insects navigate is still poorly understood. Laboratory studies suggest that locusts are able to exploit the sky polarization pattern as a navigational cue. Like other insects locusts detect polarized light through a specialized dorsal rim area (DRA) of the eye. Polarization signals are transmitted through the optic lobe to the anterior optic tubercle (AOTu) and, finally, to the central complex in the brain. Whereas neurons of the AOTu integrate sky polarization and chromatic cues in a daytime dependent manner, the central complex holds a topographic representation of azimuthal directions suggesting a role as an internal sky compass. To understand further the integration of sky compass cues we studied polarization-sensitive (POL) neurons in the medulla that may be intercalated between DRA photoreceptors and AOTu neurons. Five types of POL-neuron were characterized and four of these in multiple recordings. All neurons had wide arborizations in medulla layer 4 and most, additionally, in the dorsal rim area of the medulla and in the accessory medulla, the presumed circadian clock. The neurons showed type-specific orientational tuning to zenithal polarized light and azimuth tuning to unpolarized green and UV light spots. In contrast to neurons of the AOTu, we found no evidence for color opponency and daytime dependent adjustment of sky compass signals. Therefore, medulla layer 4 is a distinct stage in the integration of sky compass signals that precedes the time-compensated integration of celestial cues in the AOTu.     

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.     

Experience‐related reorganization of giant synapses in the lateral complex: Potential role in plasticity of the sky‐compass pathway in the desert ant Cataglyphis fortis

Cataglyphis desert ants undergo an age‐related polyethism from interior workers to relatively short‐lived foragers with remarkable visual navigation capabilities, predominantly achieved by path integration using a polarized skylight‐based sun compass and a stride‐integrating odometer. Behavioral and physiological experiments revealed that the polarization (POL) pattern is processed via specialized UV‐photoreceptors in the dorsal rim area of the compound eye and POL sensitive optic lobe neurons. Further information about the neuronal substrate for processing of POL information in the ant brain has remained elusive. This work focuses on the lateral complex (LX), known as an important relay station in the insect sky‐compass pathway. Neuroanatomical results in Cataglyphis fortis show that LX giant synapses (GS) connect large presynaptic terminals from anterior optic tubercle neurons with postsynaptic GABAergic profiles of tangential neurons innervating the ellipsoid body of the central complex. At the ultrastructural level, the cup‐shaped presynaptic structures comprise many active zones contacting numerous small postsynaptic profiles. Three‐dimensional quantification demonstrated a significantly higher number of GS (～13%) in foragers compared with interior workers. Light exposure, as opposed to age, was necessary and sufficient to trigger a similar increase in GS numbers. Furthermore, the increase in GS numbers was sensitive to the exclusion of UV light. As previous experiments have demonstrated the importance of the UV spectrum for sky‐compass navigation in Cataglyphis, we conclude that plasticity in LX GS may reflect processes involved in the initial calibration of sky‐compass neuronal circuits during orientation walks preceding active foraging. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 390–404, 2016     

Transmedulla Neurons in the Sky Compass Network of the Honeybee (Apis mellifera) Are a Possible Site of Circadian Input

Honeybees are known for their ability to use the sun’s azimuth and the sky’s polarization pattern for spatial orientation. Sky compass orientation in bees has been extensively studied at the behavioral level but our knowledge about the underlying neuronal systems and mechanisms is very limited. Electrophysiological studies in other insect species suggest that neurons of the sky compass system integrate information about the polarization pattern of the sky, its chromatic gradient, and the azimuth of the sun. In order to obtain a stable directional signal throughout the day, circadian changes between the sky polarization pattern and the solar azimuth must be compensated. Likewise, the system must be modulated in a context specific way to compensate for changes in intensity, polarization and chromatic properties of light caused by clouds, vegetation and landscape. The goal of this study was to identify neurons of the sky compass pathway in the honeybee brain and to find potential sites of circadian and neuromodulatory input into this pathway. To this end we first traced the sky compass pathway from the polarization-sensitive dorsal rim area of the compound eye via the medulla and the anterior optic tubercle to the lateral complex using dye injections. Neurons forming this pathway strongly resembled neurons of the sky compass pathway in other insect species. Next we combined tracer injections with immunocytochemistry against the circadian neuropeptide pigment dispersing factor and the neuromodulators serotonin, and γ-aminobutyric acid. We identified neurons, connecting the dorsal rim area of the medulla to the anterior optic tubercle, as a possible site of neuromodulation and interaction with the circadian system. These neurons have conspicuous spines in close proximity to pigment dispersing factor-, serotonin-, and GABA-immunoreactive neurons. Our data therefore show for the first time a potential interaction site between the sky compass pathway and the circadian clock.     

Behavioral analysis of polarization vision in tethered flying locusts

For spatial navigation many insects rely on compass information derived from the polarization pattern of the sky. We demonstrate that tethered flying desert locusts (Schistocerca gregaria) show e-vector-dependent yaw-torque responses to polarized light presented from above. A slowly rotating polarizer (5.3° s^–1) induced periodic changes in yaw torque corresponding to the 180° periodicity of the stimulus. Control experiments with a rotating diffuser, a weak intensity pattern, and a stationary polarizer showed that the response is not induced by intensity gradients in the stimulus. Polarotaxis was abolished after painting the dorsal rim areas of the compound eyes black, but remained unchanged after painting the eyes except the dorsal rim areas. During rotation of the polarizer, two e-vectors (preferred and avoided e-vector) induced no turning responses: they were broadly distributed from 0 to 180° but, for a given animal, were perpendicular to each other. The data demonstrate polarization vision in the desert locust, as shown previously for bees, flies, crickets, and ants. Polarized light is perceived through the dorsal rim area of the compound eye, suggesting that polarization vision plays a role in compass navigation of the locust.     

Evidence for instantaneous e-vector detection in the honeybee using an associative learning paradigm

Many insects use the polarization pattern of the sky for obtaining compass information during orientation or navigation. E-vector information is collected by a specialized area in the dorsal-most part of the compound eye, the dorsal rim area (DRA). We tested honeybees' capability of learning certain e-vector orientations by using a classical conditioning paradigm with the proboscis extension reflex. When one e-vector orientation (CS+) was associated with sugar water, while another orientation (CS-) was not rewarded, the honeybees could discriminate CS+ from CS-. Bees whose DRA was inactivated by painting did not learn CS+. When ultraviolet (UV) polarized light (350 nm) was used for CS, the bees discriminated CS+ from CS-, but no discrimination was observed in blue (442 nm) or green light (546 nm). Our data indicate that honeybees can learn and discriminate between different e-vector orientations, sensed by the UV receptors of the DRA, suggesting that bees can determine their flight direction from polarized UV skylight during foraging. Fixing the bees' heads during the experiments did not prevent learning, indicating that they use an 'instantaneous' algorithm of e-vector detection; that is, the bees do not need to actively scan the sky with their DRAs ('sequential' method) to determine e-vector orientation.     

Celestial orientation in bees: the use of spectral cues

Summary The spectral cues used in the bee's celestial compass are investigated by presenting bees dancing on a horizontal comb with unpolarized (or polarized) spectral stimuli. Where appropriate, the use of e-vector information is prevented by painting out the specialized dorsal margin of the bee's eye (POL area, Fig. 1). This area has been shown to mediate e-vector information (Fig. 3; Wehner 1982), whereas the remainder of the dorsal retina is sufficient for mediating spectral information (Fig. 4).Spectral cues are used by the bees to discriminate between sun and sky (Fig. 4). According to physical reality (Fig. 2), a long-wavelength stimulus is taken as the sun, whereas a short-wavelength stimulus is expected by the bee to lie anywhere within the antisolar half of the sky (Figs. 5 and 6). This is in accord with the bee's e-vector compass in which e-vectors are confined to the antisolar half of the sky (Fig. 9).In general, spectral cues do not provide precise compass information except when a full celestial colour gradient is available including the solar and the antisolar meridian (Figs. 7 and 8).     

Neural mechanisms in insect navigation: polarization compass and odometer

Insect navigation relies on path integration, a procedure by which information about compass bearings pursued and distances travelled are combined to calculate position. Three neural levels of the polarization compass, which uses the polarization of skylight as a reference, have been analyzed in orthopteran insects. A group of dorsally directed, highly specialized ommatidia serve as polarization sensors. Polarization-opponent neurons in the optic lobe condition the polarization signal by removing unreliable and irrelevant components of the celestial stimulus. Neurons found in the central complex of the brain possibly represent elements of the compass output. The odometer for measuring travelling distances in honeybees relies on optic flow experienced during flight, whereas desert ants most probably use proprioreceptive cues.     

Polarized skylight orientation in the desert antCataglyphis

Summary The desert antCataglyphis bicolor is able to use the pattern of polarized light in the sky as compass. By confronting the ant to single spots of artificially and naturally polarized light it is shown howCataglyphis uses the polarization pattern.When exposed to a horizontal e-vector,Cataglyphis was always oriented correctly. Orientation errors occurred, however, when other e-vector directions were presented. This indicates that the e-vector positions assumed by the ant do not coincide with the e-vector positions actually realized in the sky. From this it is concluded thatCataglyphis has no detailed knowledge of the actual azimuthal positions of the e-vectors. Instead, it is relying on a simplified celestial map of the polarization patterns in the sky (Fig. 7).Usually, the ant did not confuse celestial spots with identical e-vector directions. Even at sunset when the polarization pattern is completely ambiguous, correct orientation occurred. This suggests that the ant uses additional celestial cues such as the degree of polarization, the color or the intensity to find its way home when the sun is obscured.     

Ultrastructure and orientation of ommatidia in the dorsal rim area of the locust compound eye

In many insect species, a dorsal rim area (DRA) in the compound eye is adapted to analyze the sky polarization pattern for compass orientation. In the desert locust Schistocerca gregaria, these specializations are particularly striking. The DRA of the locust consists of about 400 ommatidia. The facets have an irregular shape, and pore canals are often present in the corneae. Screening pigment is missing in the region of the dioptric apparatus suggesting large receptive fields. The rhabdoms are shorter, but about four times larger in cross-section than the rhabdoms of ordinary ommatida. Eight retinula cells contribute to the rhabdom. The microvilli of retinula cell 7 and of cells 1, 2, 5, 6, 8 are highly aligned throughout the rhabdom and form two blocks of orthogonal orientation. The microvilli in the minute rhabdomeres of retinula cells 3 and 4, in contrast, show no particular alignment. As in other insect species, microvillar orientations are arranged in a fan-like pattern across the DRA. Photoreceptor axons project to distinct areas in the dorsal lamina and medulla. The morphological specializations in the DRA of the locust eye most likely maximize the polarization sensitivity and suggest that the locust uses this eye region for analysis of the sky polarization pattern.     

Navigation by light polarization in clear and turbid waters

Certain terrestrial animals use sky polarization for navigation. Certain aquatic species have also been shown to orient according to a polarization stimulus, but the correlation between underwater polarization and Sun position and hence the ability to use underwater polarization as a compass for navigation is still under debate. To examine this issue, we use theoretical equations for per cent polarization and electric vector (e-vector) orientation that account for the position of the Sun, refraction at the air-water interface and Rayleigh single scattering. The polarization patterns predicted by these theoretical equations are compared with measurements conducted in clear and semi-turbid coastal sea waters at 2 m and 5 m depth over sea floors of 6 m and 28 m depth. We find that the per cent polarization is correlated with the Sun's elevation only in clear waters. We furthermore find that the maximum value of the e-vector orientation angle equals the angle of refraction only in clear waters, in the horizontal viewing direction, over the deeper sea floor. We conclude that navigation by use of underwater polarization is possible under restricted conditions, i.e. in clear waters, primarily near the horizontal viewing direction, and in locations where the sea floor has limited effects on the light's polarization.     

Immunocytochemistry of histamine in the brain of the locust<Emphasis Type="Italic"> Schistocerca gregaria</Emphasis>

Histamine serves a neurotransmitter role in arthropod photoreceptor neurons, but is also present in a small number of interneurons throughout the nervous system. In search of a suitable model system for the analysis of histaminergic neurotransmission in insects, we mapped the distribution of histamine in the brain of the desert locust Schistocerca gregaria by immunocytochemistry. In the optic lobe, apparently all photoreceptor cells of the compound eye with projections to the lamina and medulla showed intense immunostaining. Photoreceptors of the dorsal rim area of the eye had particularly large fiber diameters and gave rise to uniform varicose immunostaining throughout dorsal rim areas of the lamina and medulla. In the locust midbrain 21 bilateral pairs of histamine-immunoreactive interneurons were found, and 13 of these were reconstructed in detail. While most neuropil areas contained a dense meshwork of immunoreactive processes, immunostaining in the antennal lobe and in the calyces of the mushroom body was sparse and no staining occurred in the pedunculus and lobes of the mushroom body, in the protocerebral bridge, and in the lower division of the central body. A prominent group of four immunostained neurons had large cell bodies near the median ocellar nerve root and descending axonal fibers. These neurons are probably identical to previously identified primary commissure pioneer neurons of the locust brain. The apparent lack in the desert locust of certain histamine-immunoreactive neurons which were reported in the migratory locust may be responsible for differences in the physiological role of histamine between both species.The study was supported by the Deutsche Forschungsgemeinschaft, grants Ho 950/13 and 950/14     

The POL area of the honey bee's eye: behavioural evidence

ABSTRACT. The uppermost dorsal part of the honey bee's compound eye contains a group of c. 150 specialized ommatidia. The photoreceptors of these ommatidia are characterized by a number of anatomical and physiological peculiarities which suggest that they have functional significance for the detection of polarized skylight. Here, we show by painting out different parts of the eye and recording the bee's behavioural responses that the specialized photoreceptors at the dorsal margin of the eye are indeed necessary for detecting polarized skylight and deriving compass information from celestial e-vector patterns. Hence, this group of specialized ommatidia can be called the POL area of the bee's compound eye.     

Coding of azimuthal directions via time-compensated combination of celestial compass cues

Many animals use the sun as a reference for spatial orientation [1-3]. In addition to sun position, the sky provides two other sources of directional information, a color gradient [4] and a polarization pattern [5]. Work on insects has predominantly focused on celestial polarization as an orientation cue [6, 7]. Relying on sky polarization alone, however, poses the following two problems: E vector orientations in the sky are not suited to distinguish between the solar and antisolar hemisphere of the sky, and the polarization pattern changes with changing solar elevation during the day [8, 9]. Here, we present neurons that overcome both problems in a locust's brain. The spiking activity of these neurons depends (1) on the E vector orientation of dorsally presented polarized light, (2) on the azimuthal, i.e., horizontal, direction, and (3) on the wavelength of an unpolarized light source. Their tuning to these stimuli matches the distribution of a UV/green chromatic contrast as well as the polarization of natural skylight and compensates for changes in solar elevation during the day. The neurons are, therefore, suited to code for solar azimuth by concurrent combination of signals from the spectral gradient, intensity gradient, and polarization pattern of the sky.     

昆虫对偏振光的响应及感受机理研究进展

偏振光是不同于普通光源的一种光, 常指光矢量在某一个方向振动的光波, 可分为线性偏振光、 圆偏振光和椭圆偏振光等。目前已经发现自然界的偏振光影响许多昆虫的行为, 如西方蜜蜂Apis mellifera的飞行导航、 蛱蝶Heliconius cydno chioneus的觅偶、 凤蝶Papilio aegeus产卵场所的选择等。金龟子对圆偏振光的反射可以作为一种分类的性状。昆虫复眼背部边缘区域（dorsal rim area, DRA）小眼是感受偏振光的主要器官, 电生理学研究表明前视神经节是蝗虫偏振视觉通路的一部分。在匈牙利, 已经开始利用偏振光研制步甲等昆虫的诱捕器。     

Distribution of PER protein,pigment-dispersing hormone,prothoracicotropic hormone,and eclosion hormone in the cephalic nervous system of insects

Investigations performed on adult insects revealed that putative components of the central pacemaker, the protein Period (PER) and the pigment-dispersing hormone (PDH), are immunocytochemically detectable in discrete sets of brain neurons throughout the class of Insecta, represented by a bristletail, mayfly, damselfly, 2 locust species, stonefly, 2 bug species, goldsmith beetle, caddisfly, honeybee, and 2 blowfly species. The PER-positive cells are localized in the frontal protocerebrum and in most species also in the optic lobes, which are their only location in damselfly and goldsmith beetle. Additional PER-positive cells occur in a few species either in the deuto- and tritocerebrum or in the suboesophageal ganglion. The PER staining was always confined to the cytoplasm. The PDH immunoreactivity consistently occurs in a cluster of perikarya located frontoventrally at the proximal edge of the medulla. The mayfly and both locust species possess additional PDH neurons in 2 posterior cell clusters at the proximal edge of the medulla, and mayfly, waterstrider, and 1 of the blowfly species in the central brain. PDH-positive fibers form a fanlike arrangement over the frontal side of the medulla. Two or just 1 bundle of PDH-positive fibers run from the optic lobe to the protocerebrum, with collaterals passing over to the contralateral optic lobe. Antisera to the prothoracicotropic (PTTH) and the eclosion (EH) hormones, which in some insects regulate the molting and ecdysis rhythms, respectively, typically react with a few neurons in the frontal protocerebrum. However, the PTTH-positive neurons of the mayfly and the damselfly and the EH-positive neurons of the caddisfly are located in the suboesophageal ganglion. No PTTH-like antigen was detected in locusts, and no EH-like antigens were detected in the damselfly, stonefly, locusts, and the honeybee. There are no signs of co-localization of the PER-, PDH-, PTTH-, and EH-like antigens in identical neurons.     

Behavioural evidence for polarization vision in crickets

ABSTRACT. Tethered field crickets, Gryllus campestris L., walking on an air-suspended bail exhibit a spontaneous response to the e-vector of polarized light presented from above: E-vector orientation controls strength and direction of turning tendency. Experiments in which different eye regions are covered with paint suggest that this response is mediated by the anatomically and physiologically specialized dorsal rim area of the compound eye. We conclude that crickets have polarization vision and that the dorsal rim area of the eye plays a key role in this sensory capacity.