Elke K. Buschbeck

Alterations of the CIB2 calcium- and integrin-binding protein cause Usher syndrome type 1J and nonsyndromic deafness DFNB48

Sensorineural hearing loss is genetically heterogeneous. Here, we report that mutations in CIB2, which encodes a calcium- and integrin-binding protein, are associated with nonsyndromic deafness (DFNB48) and Usher syndrome type 1J (USH1J). One mutation in CIB2 is a prevalent cause of deafness DFNB48 in Pakistan; other CIB2 mutations contribute to deafness elsewhere in the world. In mice, CIB2 is localized to the mechanosensory stereocilia of inner ear hair cells and to retinal photoreceptor and pigmented epithelium cells. Consistent with molecular modeling predictions of calcium binding, CIB2 significantly decreased the ATP-induced calcium responses in heterologous cells, whereas mutations in deafness DFNB48 altered CIB2 effects on calcium responses. Furthermore, in zebrafish and Drosophila melanogaster, CIB2 is essential for the function and proper development of hair cells and retinal photoreceptor cells. We also show that CIB2 is a new member of the vertebrate Usher interactome.

Evolution of Insect Eyes: Tales of Ancient Heritage, Deconstruction, Reconstruction, Remodeling, and Recycling

The visual organs of insects are known for their impressive evolutionary conservation. Compound eyes built from ommatidia with four cone cells are now accepted to date back to the last common ancestor of insects and crustaceans. In species as different as fruit flies and tadpole shrimps, the stepwise cellular patterning steps of the early compound eye exhibit detailed similarities implying 500 million years of developmental conservation. Strikingly, there is also a cryptic diversity of insect visual organs, which gives proof to evolution’s versatility in molding even the most tenacious structures into something new. We explore this fascinating aspect in regard to the structure and function of a variety of different insect eyes. This includes work on the unique compound–single-chamber combination eye of twisted-winged insects and the bizarre evolutionary trajectories of specialized larval eyes in endopterygote insects.

The unusual visual system of the Strepsiptera: external eye and neuropils

Adult males of the insect order Strepsiptera are characterized by an unusual visual system that may use design principles from compound as well as simple eyes. The lenses of this eye are unusually large and focus images onto extended retinae. The light-gathering ability of the lens is sufficient to resolve multiple points of an image in each optical unit. We regard each unit as an independent image-forming eye that contributes an inverted partial image. Each partial image is re-inverted by optic chiasmata between the retinae and the lamina, where the complete image could be assembled from the neighboring units. The lamina, medulla and lobula are present, but their organization into cartridges is not clearly discernable. Fluorescent fills, whole-tissue stains, and synaptotagmin immunohistochemistry show that the optic neuropils nevertheless are densely packed, and that several parallel channels within the medulla underlie each of the lenses. The size and shape of the rhabdoms, as well as a relatively slow flicker-fusion frequency could suggest that these eyes evolved through a nocturnal life stage.

Twenty-eight retinas but only twelve eyes: an anatomical analysis of the larval visual system of the diving beetle Thermonectus marmoratus (Coleoptera: Dytiscidae).

The larvae of the sunburst diving beetle, Thermonectus marmoratus (Coleoptera: Dytiscidae), are highly efficient visually guided predators. Their visual system consists of a cluster of six stemmata and one eye patch on each side of the head capsule. Histological investigations show that the organization of individual stemmata differs strongly from any eye that has previously been described. Based on general morphology, ultrastructural data, and the presence of actin‐rich areas that are typical for rhabdoms, we find that each eye is characterized by several retinas. The most dorsal eye on each side is relatively long and tubular, and we have identified three spatially distinct areas with retinula cells: 1) a band of two rows of rhabdoms along the medial side of the eye tube; 2) a flattened cone‐shaped region towards the bottom of the tube that is formed by the rhabdoms of retinula cells that are oriented perpendicular to the light path; and 3) two horizontal rows of long rhabdoms parallel to the light path at the base of the tube. A second large eye is organized similarly but lacks the medial band. The remaining four eyes are nearly spherical and each has two distinct retinas. The 12 eyes hence account for a total of 26 retinas, and two further retinas are present in eye patches lacking lenses. Our anatomical findings suggest that this is an example of a visual system in which specific visual tasks are distributed among the eyes, and which relies on a variety of highly specialized retinas. J. Comp. Neurol. 497:166–181, 2006.

Spatial distribution of opsin-encoding mRNAs in the tiered larval retinas of the sunburst diving beetle Thermonectus marmoratus (Coleoptera: Dytiscidae)

SUMMARY Larvae of the sunburst diving beetle, Thermonectus marmoratus, have a cluster of six stemmata (E1-6) and one eye patch on each side of the head. Each eye has two retinas: a distal retina that is closer to the lens, and a proximal retina that lies directly underneath. The distal retinas of E1 and E2 are made of a dorsal and a ventral stack of at least twelve photoreceptor layers. Could this arrangement be used to compensate for lens chromatic aberration, with shorter wavelengths detected by the distal layers and longer wavelengths by the proximal layers? To answer this question we molecularly identified opsins and their expression patterns in these eyes. We found three opsin-encoding genes. The distal retinas of all six eyes express long-wavelength opsin (TmLW) mRNA, whereas the proximal retinas express ultraviolet opsin (TmUV I) mRNA. In the proximal retinas of E1 and E2, the TmUV I mRNA is expressed only in the dorsal stack. A second ultraviolet opsin mRNA (TmUV II), is expressed in the proximal retinas of E1 and E2 (both stacks). The finding that longer-wavelength opsins are expressed distally to shorter-wavelength opsins makes it unlikely that this retinal arrangement is used to compensate for lens chromatic aberration. In addition, we also described opsin expression patterns in the medial retina of E1 and in the non-tiered retina of the lensless eye patch. To our knowledge, this is also the first report of multiple UV opsins being expressed in the same stemma.

Scanning behavior by larvae of the predacious diving beetle, Thermonectus marmoratus (Coleoptera: Dytiscidae) enlarges visual field prior to prey capture

Larvae of the predaceous diving beetle Thermonectus marmoratus bear six stemmata on each side of their head, two of which form relatively long tubes with linear retinas at their proximal ends. The physical organization of these eyes results in extremely narrow visual fields that extend only laterally in the horizontal body plane. There are other examples of animals possessing eyes with predominantly linear retinas, or with linear arrangements of specific receptor types. In these animals, the eyes, or parts of the eyes, are movable and perform scanning movements to increase the visual field. Based on anatomical data and observations of relatively transparent, immobilized young larvae, we report here that T. marmoratus larvae are incapable of moving their eyes or any part of their eyes within the head capsule. However, they do perform a series of bodily dorso-ventral pivots prior to prey capture, behaviorally extending the vertical visual field from 2° to up to 50°. Frame-by-frame analysis shows that such behavior is performed within a characteristic distance to the prey. These data provide first insights into the function of the very peculiar anatomical eye organization of T. marmoratus larvae.

Spectral sensitivity of the principal eyes of sunburst diving beetle, Thermonectus marmoratus (Coleoptera: Dytiscidae), larvae.

SUMMARY The principal eyes of sunburst diving beetle, Thermonectus marmoratus, larvae are among the most unusual eyes in the animal kingdom. They are composed of long tubes connecting bifocal lenses with two retinas: a distal retina situated a few hundred micrometers behind the lens, and a proximal retina that is situated directly beneath. A recent molecular study on first instar larvae suggests that the distal retina expresses a long-wavelength-sensitive opsin (TmLW), whereas the proximal retina predominantly expresses an ultraviolet-sensitive opsin (TmUV II). Using cloning and in situ hybridization we here confirm that this opsin distribution is, for the most part, maintained in third instar larvae (with the exception of the TmUV I that is weakly expressed only in proximal retinas of first instar larvae). We furthermore use intracellular electrophysiological recordings and neurobiotin injections to determine the spectral sensitivity of individual photoreceptor cells. We find that photoreceptors of the proximal retina have a sensitivity curve that peaks at 374–375 nm. The shape of the curve is consistent with the predicted absorbance of a single-opsin template. The spectral response of photoreceptors from the distal retina confirms their maximum sensitivity to green light with the dominant λ-peak between 520 and 540 nm, and the secondary β-peak between 340 and 360 nm. These physiological measurements support molecular predictions and represent important steps towards understanding the functional organization of the unusual stemmata of T. marmoratus larvae.

Eye stalks or no eye stalks: A structural comparison of pupal development in the stalk-eyed fly cyrtodiopsis and in drosophila

After emergence from the puparium, stalk‐eyed flies of the family Diopsidae rapidly expand their head capsule so that the eyes and optic lobes are displaced at the ends of stalks that extend from the central head. Because the expansion takes place in only 15 minutes, we are especially interested in ontogenetic modifications that may facilitate such a rapid and dramatic change. To examine the pupal development of the brain, we used Bodian staining in the stalk‐eyed fly, Cyrtodiopsis whitei and compared it with development in the fruit fly, Drosophila melanogaster, which serves as a “typical” dipteran example without eye stalks. Early in pupal development, the neuropil organization of the two species is fairly similar. In both species, columns are present in the outer medulla and giant fibers are discernible in the lobula plate. In contrast to D. melanogaster, C. whitei shows a small, neck‐like constriction between the optic lobes and the rest of the brain. By 20% of pupal development, the divergence is more apparent, and by 30%, the future eye stalk and optic nerve of C. whitei has started to form. During the remaining 70% of development, the initially thick optic nerve narrows, and becomes gradually elongated, eventually coiling and folding throughout the short eye stalk. Similarly, the cuticle of the surrounding region becomes constricted, slightly elongated, and gradually appears more and more densely corrugated, like an accordion bellows. However, except for the formation of the optic nerve, the dense aggregation of cuticle around it, and a shift in orientation of the neuropils, the developmental programs of the two species are remarkably similar. This suggests that only a few aspects of development have been modified during the course of evolution to generate the stalk‐eyed phenotype. At eclosion, the imago of C. whitei goes through a pumping process to inflate the eye stalks to their full length. Measurements of the diameter of the optic nerve before and after the expansion reveal only a small decrease. We propose that the cuticular folding of the eye stalk as well as the coiling of the optic nerve prepare the pupa well for the rapid and dramatic eye‐stalk inflation after eclosion. J. Comp. Neurol. 433:486–498, 2001.

NEUROBIOLOGICAL CONSTRAINTS AND FLY SYSTEMATICS: HOW DIFFERENT TYPES OF NEURAL CHARACTERS CAN CONTRIBUTE TO A HIGHER LEVEL DIPTERAN PHYLOGENY

Abstract.— Much uncertainty still exists regarding higher level phylogenetic relationships in the insect order Diptera, and the need for independent analyses is apparent. In this paper, I present a parsimony analysis that is based on details of the nervous system of flies. Because neural characters have received little attention in modern phylogenetic analyses and the stability of neural traits has been debated, special emphasis is given to testing the robustness of the analysis itself and to evaluating how neurobiological constraints (such as levels of neural processing) influence the phylogenetic information content.

Behavioral evidence for within-eyelet resolution in twisted-winged insects (Strepsiptera)

SUMMARY Compound eyes are typically composed of hundreds to thousands of ommatidia, each containing 8–10 receptors. The maximal spatial frequency at which a compound eye can sample the environment is determined by the inter-ommatidial angle. Males of the insect order Strepsiptera are different: their eyes are composed of a smaller number of relatively large units (eyelets), each with an extended retina. Building on a study of Xenos vesparum, we use a behavioral paradigm based on the optomotor response to investigate the possibility that the eyelets of the Strepsiptera Xenos peckii are image-forming units. From anatomical evidence, we hypothesize that spatial sampling in the strepsipteran eye is determined not only by the interactions of widely spaced photoreceptors in different eyelets, but also by the angular separation between groups of closely spaced photoreceptors within eyelets. We compared X. peckiis optomotor response with the predictions of an elementary motion detector (EMD) model consisting of two distinctly different sampling bases. The best match between our empirical results and the model shows that the optomotor response in X. peckii males is determined by both the small (intra-eyelet) and large (possibly inter-eyelet) separations. Our results indicate that the X. peckii eye has sampling bases around 10° and 20°, and that each eyelet could be composed of up to 13 sampling points, which is consistent with previous anatomical findings. This study is the first to use the EMD model explicitly to investigate the possibility that strepsipteran eyes combine motion detection features from both camera and compound eyes.