Gerta Fleissner

Ultrastructural analysis of a putative magnetoreceptor in the beak of homing pigeons

With the use of different light and electron microscopic methods, we investigated the subcellular organization of afferent trigeminal terminals in the upper beak of the homing pigeon, Columba livia, which are about 5 μm in diameter and contain superparamagnetic magnetite (SPM) crystals. The SPM nanocrystals are assembled in clusters (diameter, ∼1–2 μm). About 10 to 15 of these clusters occur inside one nerve terminal, arranged along the cell membrane. Each SPM cluster is embedded in a solid fibrous cup, open towards the cell surface, to which the cluster adheres by delicate fiber strands. In addition to the SPM clusters, a second inorganic iron compound has been identified: noncrystalline platelets of iron phosphate (about 500 nm wide and long and maximally 100 nm thick) that occur along a fibrous core of the terminal. The anatomic features suggested that these nerve endings could detect small intensity changes of the geomagnetic field. Such stimuli can induce deformations of the SPM clusters, which could be transduced into primary receptor potentials by mechanosensitive membrane receptor channels. The subepidermal fat cells surrounding the nerve endings prevent the inside from external mechanical stimuli. These structural findings corresponded to conclusions inferred from rock magnetic measurements, theoretical calculations, model experiments, and behavioral data, which also matched previous electrophysiologic recordings from migratory birds. J. Comp. Neurol. 458:350–360, 2003.

A novel concept of Fe-mineral-based magnetoreception: histological and physicochemical data from the upper beak of homing pigeons

Animals make use of the Earth’s magnetic field for navigation and regulation of vegetative functions; however, the anatomical and physiological basis for the magnetic sense has not been elucidated yet. Our recent results from histology and X-ray analyses support the hypothesis that delicate iron-containing structures in the skin of the upper beak of homing pigeons might serve as a biological magnetometer. Histology has revealed various iron sites within dendrites of the trigeminal nerve, their arrangement along strands of axons, the existence of three dendritic fields in each side of the beak with specific 3D-orientations, and the bilateral symmetry of the whole system. Element mapping by micro-synchrotron X-ray fluorescence analysis has shown the distribution of iron and its quantities. Micro-synchrotron X-ray absorption near-edge-structure spectroscopy has allowed us to unambiguously identify maghemite as the predominating iron mineral (90 vs 10% magnetite). In this paper, we show that iron-based magnetoreception needs the presence of both of these iron minerals, their specific dimensions, shapes, and arrangements in three different subcellular compartments. We suggest that an inherent magnetic enhancement process via an iron-crusted vesicle and the attached chains of iron platelets might be sufficient to account for the sensitivity and specificity required by such a magnetoreceptor. The appropriate alignment between the Earth’s magnetic field and the maghemite bands would induce a multiple attraction of the magnetite bullets perpendicular to the membrane, thus, triggering strain-sensitive membrane channels and a primary receptor potential. Due to its 3D architecture and physicochemical nature, the dendritic system should be able to separately sense the three vector components of the Earth’s local field, simultaneously—allowing birds to detect their geographic position by the magnetic vector, i.e., amplitude and direction of the local magnetic field, irrespective of the animal’s posture or movement and photoreception.

Avian Ultraviolet/Violet Cones Identified as Probable Magnetoreceptors

Background The Radical-Pair-Model postulates that the reception of magnetic compass directions in birds is based on spin-chemical reactions in specialized photopigments in the eye, with cryptochromes discussed as candidate molecules. But so far, the exact subcellular characterization of these molecules in the retina remained unknown. Methodology/Principal Findings We here describe the localization of cryptochrome 1a (Cry1a) in the retina of European robins, Erithacus rubecula, and domestic chickens, Gallus gallus, two species that have been shown to use the magnetic field for compass orientation. In both species, Cry1a is present exclusively in the ultraviolet/violet (UV/V) cones that are distributed across the entire retina. Electron microscopy shows Cry1a in ordered bands along the membrane discs of the outer segment, and cell fractionation reveals Cry1a in the membrane fraction, suggesting the possibility that Cry1a is anchored along membranes. Conclusions/Significance We provide first structural evidence that Cry1a occurs within a sensory structure arranged in a way that fulfils essential requirements of the Radical-Pair-Model. Our findings, identifying the UV/V-cones as probable magnetoreceptors, support the assumption that Cry1a is indeed the receptor molecule mediating information on magnetic directions, and thus provide the Radical-Pair-Model with a profound histological background.

Avian Magnetoreception: Elaborate Iron Mineral Containing Dendrites in the Upper Beak Seem to Be a Common Feature of Birds

The magnetic field sensors enabling birds to extract orientational information from the Earths magnetic field have remained enigmatic. Our previously published results from homing pigeons have made us suggest that the iron containing sensory dendrites in the inner dermal lining of the upper beak are a candidate structure for such an avian magnetometer system. Here we show that similar structures occur in two species of migratory birds (garden warbler, Sylvia borin and European robin, Erithacus rubecula) and a non-migratory bird, the domestic chicken (Gallus gallus). In all these bird species, histological data have revealed dendrites of similar shape and size, all containing iron minerals within distinct subcellular compartments of nervous terminals of the median branch of the Nervus ophthalmicus. We also used microscopic X-ray absorption spectroscopy analyses to identify the involved iron minerals to be almost completely Fe III-oxides. Magnetite (Fe II/III) may also occur in these structures, but not as a major Fe constituent. Our data suggest that this complex dendritic system in the beak is a common feature of birds, and that it may form an essential sensory basis for the evolution of at least certain types of magnetic field guided behavior.

Clusters of superparamagnetic magnetite particles in the upper-beak skin of homing pigeons evidence of a magnetoreceptor?

Previous electrophysiological studies on bobolinks, an American migratory songbird, and on homing pigeons suggested that the skin of the upper beak may be involved in magnetic-field perception, which makes this tissue likely to contain a magnetic-field receptor. In the upper-beak skin of homing pigeons, we localised high concentrations of Fe3+, which form distinct coherent elongated structures extending up to 200 μ in length. Rather than being randomly distributed over the tissue, these structures always occur in the same skin layer, the stratum laxum of the subcutis. Using transmission electron microscopy (TEM), we identified the material as aggregates of magnetite (Fe3O4) nanocrystals, in the grain-size range of superparamagnetism at ambient temperatures. The nanocrystals (with grain-sizes between 2 and 5 nm) form densely packed, encapsulated clusters of 1 to 3 μ in diameter. It is demonstrated that such a cluster undergoes shape changes as the magnetic field changes, and thus could represent the core of a magnetic-field receptor. This interpretation is supported by the fact that the found structures are adjacent to nervous material. This paper was presented at the “Biogenic Iron Minerals” symposium held in Tihany, Hungary (May 2000)

The optic nerve mediates the circadian pigment migration in the median eyes of the scorpion

Abstract 1. 1. The peripheral visual pathway from the median eyes of the scorpion Androctonus australis was interrupted at different points and the effect on the circadian rhythm of median-eye sensitivity was examined. 2. 2. Any interruption of the visual pathway distal to the supraesophageal ganglion abolishes the circadian sensitivity rhythm in the median eyes. This rhythm is thus controlled by efferents in the optic nerve (very probably via the neurosecretory axons) rather than by way of the hemolymph. 3. 3. Following transection of the optic nerve, the sensitivity of the median eyes proceeds rapidly to the daytime state. This condition is associated with movement of the screening pigment into the distal ends of the visual cells. 4. 4. The oscillator system controlling the circadian pigment migration in the median eye cannot be located in the eye itself, but must lie in the CNS, proximal to the first optic ganglion. The oscillator itself need not be connected to both median eyes in order to function normally, as revealed by the continued rhythm in the contralateral eye following unilateral optic nerve section.

A new type of putative non-visual photoreceptors in the optic lobe of beetles

A putative photoreceptor organ is described in the carabid beetle, Pachymorpha sexguttata. The elongated structure, about 20–40 μm wide and more than 300 μm long, is situated within the optic lobe at the fronto-dorsal rim of the lamina. It lies, deep in the head capsule, in front of the compound eyes and beneath window-like thinnings of the cuticle. The organ is composed of two types of cells: (1) clear sheath cells and (2) well-organized inner receptor cells that appear in a horseshoe-like or circular array in cross-section. Common histological features of all inner cells include a distal trunk ending in microvilli that form a rhabdom-like structure, an axon at the proximal end of the cell, lamellar and multivesicular bodies within the trunk, and clusters of small mitochondria. The organ has no shielding pigment. It is connected by thin axons to a circumscribed neuropil that parallels the organ, and thence via a fiber tract to the medulla accessoria, a possible site of the circadian pacemaker in insects. Immunoreactivity to anti-pers, an antibody recognizing the Drosophila period (per) protein that plays a central role in the function of the circadian pacemaker in fruit flies, is demonstratable in thin efferent terminals within the organ, in the associated neuropil and in its fiber connection to the medulla. A second receptor organ displaying the same fine structure lies near the second optic chiasm. This set of putative photoreceptors also occurs in the tenebrionid beetle, Zophobas morio, and its pupa. The possible function of these receptor organs is discussed with respect to former chronobiological data and some recently described types of extraretinal photoreceptors in arthropods.

Staining in the brain of Pachymorpha sexguttata mediated by an antibody against a Drosophila clock-gene product: labeling of cells with possible importance for the beetle’s circadian rhythms

Abstract.Central nervous system ganglia within the head of the beetle Pachymorpha sexguttata were labeled using an antibody that recognizes an evolutionarily conserved region of the period (per) gene product of Drosophila melanogaster. per and the protein it encodes (PER) are believed to play a central role in the generation of endogenous circadian rhythms in flies; therefore anti-PER-mediated immunoreactivity may help to uncover cellular components of the circadian clock system in that insect and in others. In the beetle, application of this antibody led to the staining of a distinct set of neurons located in the optic lobes and the central brain, plus small numbers of putative glial cells in the optic lobes. Neuronal perikarya (including their nuclei in a few cases), the axons, and terminal regions of the neurons were stained. The network formed by these labeled cells and processes are candidates for the neuronal basis of the beetle’s circadian clock system: the pacemaker region situated next to the medulla neuropil, its connection to the apparent site of Zeitgeber input, and putative efferent pathways projecting to control centers of various effector systems. Anti-PER-mediated labeling and that resulting from application to beetle specimens of an antiserum against pigment-dispersing hormone (PDH) were compared; in the Drosophila brain all ’’PDH cells’’ express the per gene as well. In the beetle, however, the set of ’’PER cells’’ and PDH ones is at least in part nonoverlapping. The hypothesis that neurons stained by application of anti-PER participate in the control of the beetle’s circadian rhythms is discussed in the context of previous electrophysiological and immunohistochemical studies. Also considered are analogies to, and differences from, labeling of the PER protein in fruit flies and PER-like immunoreactivity in other animals.

Neurobiology of a Circadian Clock in the Visual System of Scorpions

What does a circadian clock look like in terms of its anatomy and physiology? This is a challenging question for neurobiologists, on which we have been focusing our research for the past several years.

Localization of components of the renin-angiotensin system in the suprachiasmatic nucleus of normotensive Sprague-Dawley rats: part A. angiotensin I/II, a light and electron microscopic study.

The central pacemaker of the mammalian circadian clock, identified in the suprachiasmatic nucleus (SCN), is of special interest for many chronomedical studies on neuropeptides. Based on its role in the modulation of blood pressure and vasopressin release, the distribution and function of the neuropeptide angiotensin II (ANG II) in the SCN became a target for several immunohistological studies. At the light microscopic level, the distribution of ANG II in the SCN is well known, but detailed information about the localization of ANG II in the SCN at the ultrastructural level is missing. To gain further insight in the functional aspects of ANG II in the SCN, we investigated on the subcellular localization of the neuropeptide ANG II and its precursor ANG I in the SCN. The current report presents a light and electron microscopic study on ANG I/II-immunoreactivity in the suprachiasmatic nucleus of normotensive Sprague-Dawley rats.