Thesis (Ph. D.)--University of Virginia, 1999. / Spine title: Structure - function analysis of IRBP. Includes bibliographical references (p. 167-172). Also available online through Digital Dissertations.

Sight in subterranean mammals living in a dark ecotope has generally been assumed as not needed and therefore greatly diminished in its function. Recent neuroanatomical studies demonstrate unexpected preservation of the visual system of several African mole-rats (Bathyergidae, Rodentia). Only a few behavioural studies, testing visual abilities and discussing their adaptive significance in these rodents, have been published to date. A spontaneous preference to light stimuli of two mole-rat species, the silvery mole-rat (Heliophobius argenteocinereus) and the giant mole-rat (Fukomys mechowii), was tested in this study. Assessed results showed convincingly that both species are able to perceive light. The following experiments provided the first behavioural support to the perception of short-wavelengths in this intensively studied group of subterranean rodents.

Retinal diseases such as age-related macular degeneration (AMD) are the major cause of blindness in the developed world. Early diagnosis of these diseases is difficult as symptoms appear only at advanced stages. Nevertheless, several studies suggest that impairment of dark adaptation (the ability of the retina to adapt to low lighting) is a cue to AMD. Dark adaptation is the result of the regeneration of light sensitive pigments after having reacted to light (bleaching). This PhD aims at developing a tool for objective measurements of the quantity of photopigment and the kinetics of dark adaptation. This work comprises a thorough review of the absorbing and reflecting properties of the different ocular structures, giving rise to a new model of retinal (or fundus) reflectance. This model provides a detailed description of the different pathways of light through the photoreceptor layer and was able to explain measurements and findings of the literature, in particular the effect of the photoreceptor matrix interstices. An extensive study of the influence of the different model parameters on the total fundus reflectance led to the proposal of a new objective and comparable measure of quantity of photopigment (QoP). This measure is obtained by fitting a constrained version of the new model to a double density difference (DDD) measurement (i.e. the logarithmic difference between reflectances of a retinal area in bleached and dark adapted states). This approach was validated by correctly fitting several DDD measurements from the literature. Future experimental studies are needed to confirm the relevance of the new QoP measure and specify its application in clinical diagnosis. Several fundus reflectometry instruments have been able to measure the DDD in human eyes however because of practical and technological limitations none of these instruments were suitable for clinical use. Here, these limitations are discussed and two new imaging fundus reflectometers are presented. Developed respectively by 4D Optics Ltd. and the Vision Research group at the University of Manchester, these two systems, based on modified fundus cameras, are ongoing development work towards clinically suitable imaging fundus reflectometry. Example data obtained with these two instruments exhibits aberrant points and low signal to noise ratio (SNR). The main issues encountered were camera noise and stability, uneven retinal illumination, and subject’s eye movements and changes of alignment. It is believed that these issues can be overcome with current technologies. One important impediment to the use of the dark adaptation experiment in clinical practice is the time it takes for photopigments to completely regenerate (up to 40 min in normal eyes). A theoretical data analysis strategy using the new model of fundus reflectance and the Marhoo, Lamb and Pugh model of photopigment regeneration kinetics is proposed to rapidly diagnose an abnormal regeneration, hence reducing considerably the duration of the experiment. This idea has not been tested on experimental data but may become relevant once better quality measurements of DDD are obtained.

L’imagerie super-résolue délivrée par l’optique adaptative permet d’imager la rétine à l’échelle cellulaire et rentre au service du diagnostic et du suivi de pathologie de l’oeil mais aussi à une meilleure connaissance de l’anatomie, des fonctions et des mécanismes de la rétine.Dans un premier temps nous avons défini la méthode permettant de positionner le centre de la fovéa ainsi que de mesurer la densité des cônes avec la meilleure répétabilité. Le centre de l’ellipse de plus fortes densités permet de positionner le centre de la fovéa. La densité des cônes est mesurée dans une fenêtre de 80 par 80 pixels à une excentricité mesurée sur un montage établi à partir de clichés réalisés tous les 2°.Un des aspects fondamental de ce projet est la réalisation d’une base de données cliniques normatives liées à des bio-marqueurs (densité, espacement et morphologie des cônes). En effet il est nécessaire de fournir des mesures quantitatives des structures visualisées. Cette étude a permis de caractériser la structure rétinienne d’une population saine ne présentant aucune maladie rétinienne, de façon à établir un référentiel de normalité. Cette étude nous a aussi permis de répondre aux questions laissées sans réponse par la littérature. Nous avons constaté que la densité des cônes différait nettement entre les méridiens horizontaux et verticaux. Nous avons également observé que la longueur axiale influait sur la densité des cônes exprimée en unité de surface mais pas en terme d'angle visuel, supportant l'hypothèse que la rétine s'étirait avec l'allongement du globe oculaire. Enfin, l'âge ne semble pas avoir d'impact sur la densité des cônes excepté à 2°.Nous nous sommes ensuite intéressé au lien entre l’acuité visuelle périphérique et l’espacement des cônes. Il est connu que les cellules ganglionnaires midgets (mRGC) sont responsables de la résolution visuelle. Mais, au niveau de la fovéa, chaque cône se connecte, via des cellules bipolaires, à une mRGC ON et OFF. Cette particularité anatomique rend la mesure de l’espacement entre les cônes un substitut observable aux cellules ganglionnaires permettant ainsi le lien avec l'acuité visuelle. Nous avons donc déterminé jusqu'à quelle excentricité les cônes ne sont plus un substitut observable des cellules ganglionnaires. Nous avons constaté que l’acuité visuelle est régit par l’espacement des cônes jusqu’à 2° d’excentricité, à la fois en rétine nasale et inférieure, soutenant l'idée que seulement 50% des mRGC fovéale détermine l’acuité visuelle (mRGC ON ou OFF). Au delà, l’espacement des cônes surestime l’acuité visuelle. Watson a développé un modèle permettant de prédire l’acuité visuelle sur la base de l’espacement entre les mRGC lui-même dérivé de l’espacement entre les cônes. Le modèle 50% (mRGC ON ou OFF) prédit également l’acuité visuelle dans le méridien vertical (inférieur) à 4° et 6° d'excentricité. Le long du méridien nasal, le modèle 50% sous-estime l’acuité visuelle. Un chevauchement partiel des champs récepteurs des mRGC ON + OFF pourrait en être la cause. Nous avons ensuite mesuré la densité des cônes dans les 48° centraux le long du méridien horizontal dans deux groupes, l’un présentant une histoire d’évolution myopique (i.e. allongement de l'oeil) et l'autre non. À ce jour, il n'existe qu’une seule étude ayant des mesures in-vivo de la densité des cônes au delà des 15° centraux. Cette étude nous a également permis d'affirmer que lors de l'évolution myopique, la rétine s’étirerait de manière uniforme dans la zone mesurée, excepté proche de la tête du nerf optique, où il semblerait qu'il y ait une adhérence plus forte. / The high-resolved imagery delivered by the adaptive optic is used to form pictures of the retina at the cellular level and to diagnose and follow up on eyes pathologies, but also to improve knowledge of the anatomy, functions and mechanisms of the retina.First we defined the method that allows us to position the center of the fovea and to measure the density of the cones with the best repeatability. The center of the ellipse of higher densities makes it possible to position the center of the fovea. The cone density is measured in a region of interest of 80 x 80 pixels to an eccentricity measured on a montage established from pictures made every 2°.One of the fundamental aspects of this project is the creation of a normative clinical database linked to bio-markers (density, spacing and morphology of cones). Indeed it is necessary to provide quantitative measures of the structures visualized. This study made it possible to characterize the retinal structure of a healthy population with no retinal disease, so as to establish a reference of normality. This study also allowed us to answer questions left unanswered by the literature. The density of the cones differs sharply between the horizontal and vertical meridians. We have also observed that the axial length influences cone density expressed in metric unit but not in terms of visual angle, supporting the hypothesis that the retina is stretched with the lengthening of the eyeball. Finally, age does not seem to have an impact on cone density except at 2°.We then examined the relationship between peripheral visual acuity and cone spacing. It is known that midget ganglion cells are in charge of visual resolution. But, at the fovea, each cone connects, via bipolar cells, to a midget ganglion cell ON and OFF. This anatomical particularity makes the measurement of the cone spacing an observable substitute for ganglion cells thus allowing the link with visual acuity. We have therefore determined to what eccentricity the cones are no longer an observable substitute for ganglion cells. We found that visual acuity is governed by the arrangement of cones up to 2° of eccentricity, both in nasal and inferior retinas, supporting the idea that only 50% of foveal mRGCs determine VA (mRGC ON or OFF). Beyond this, cone spacing over-samples ganglion cells. Watson developed a model to predict visual acuity based on the mRGC spacing, itself derived from the cone spacing. The 50% model (mRGC ON or OFF) also predicts visual acuity in the inferior meridian at 4° and 6° of eccentricity. Along the nasal meridian, the 50% model underestimates visual acuity. Partial overlap of the mRGC ON + OFF receptive fields may be the cause.Finally, we measured the cone density in the central 48° along the horizontal meridian in two study groups, one with a history of myopic evolution (i.e. elongation of the eye) and the other without. To this date, there is only one study with in-vivo measurements of cone density beyond the central 15°. This study also allowed us to assert that during myopic evolution, the retina stretches uniformly in the measured area, excepted near the optic nerve head, where it appears that there is a stronger adhesion.

Color vision is widespread in the animal kingdom, and describes the ability to discriminate between objects purely based on the wavelengths that they reflect. Experiments across many species have isolated wavelength comparison in the brain as a computation underlying color vision. This comparison takes place in color opponent neurons, which respond with opposite polarity to wavelengths in different parts of the spectrum. In this work, I explore color opponency in the genetically tractable organism Drosophila melanogaster, where these circuits have only just begun to be described. Using two-photon calcium imaging, I measure the spectral tuning of photoreceptors in the fruit fly and identify circuit mechanisms that give rise to opponency. I find two pathways: an insect-specific pathway that compares wavelengths at each point in space, and a horizontal-cell-mediated pathway similar to that found in mammals. The horizontal-cell-mediated pathway enables additional spectral comparisons through lateral inhibition, expanding the range of chromatic encoding in the fly. Together, these two pathways enable efficient decorrelation and dimensionality reduction of photoreceptor signals while retaining maximal chromatic information. This dual mechanism combines motifs of both an insect-specific visual circuit and an evolutionarily convergent circuit architecture, endowing flies with the ability to extract chromatic information at distinct spatial resolutions.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20804号 / 医博第4304号 / 新制||医||1025(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 河本 宏, 教授 松田 文彦, 教授 柳田 素子 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

No description available.

Phosducin (Pdc) is a 28 kDa binding partner for the G protein beta gamma subunit dimer (G-beta-gamma) found abundantly in the photoreceptor cells of the retina and pineal gland. In the retina, light-dependent changes in cAMP and Ca2+ control the phosphorylation of Pdc at serine 73 and 54, respectively, which in turn controls the binding of Pdc to G protein beta gamma subunit dimer . G protein beta gamma subunit dimer binding has been proposed to facilitate light-driven transport of G protein beta gamma subunit dimer from the site of phototransduction in the outer segment of the photoreceptor cell to the inner segment, thereby decreasing light sensitivity and contributing to the process of light adaptation. Dopamine and melatonin are neuromodulators whose concentrations in the retina vary reciprocally during the circadian cycle, with dopamine high during the day and melatonin high during the night. Together, they control numerous aspects of light and dark adaptation in the retina. In this study, we have investigated the possible roles of dopamine and melatonin in regulating Pdc phosphorylation. Using phosphorylation-site specific antibodies to serines 54 and 73, we show that dopamine decreases the phosphorylation of both sites. This decrease is blocked by D4 receptor antagonists and pertussis toxin, indicating that dopamine causes a decrease in photoreceptor cell cAMP and Ca2+ concentration via the D4 receptor coupled to the Gi protein. Conversely, melatonin increases the phosphorylation of both S54 and S73, most likely via the inhibition of dopamine synthesis. These results demonstrate that dopamine and melatonin control the phosphorylation state of phosducin by changing the concentration of cAMP and Ca2+ in photoreceptor cells, and they suggest that dopamine and melatonin may contribute to the light-induced movement of the photoreceptor G protein by regulating Pdc phosphorylation.

Diss. (sammanfattning) Stockholm : Karol. inst., 2006. / Härtill 6 uppsatser.

Thesis (doctoral)--Lund University, 1997. / Added t.p. with thesis statement inserted. Includes bibliographical references.