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      Selective Stimulation of Penumbral Cones Reveals Perception in the Shadow of Retinal Blood Vessels

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          Abstract

          In 1819, Johann Purkinje described how a moving light source that displaces the shadow of the retinal blood vessels to adjacent cones can produce the entopic percept of a branching tree. Here, we describe a novel method for producing a similar percept. We used a device that mixes 56 narrowband primaries under computer control, in conjunction with the method of silent substitution, to present observers with a spectral modulation that selectively targeted penumbral cones in the shadow of the retinal blood vessels. Such a modulation elicits a clear Purkinje-tree percept. We show that the percept is specific to penumbral L and M cone stimulation and is not produced by selective penumbral S cone stimulation. The Purkinje-tree percept was strongest at 16 Hz and fell off at lower (8 Hz) and higher (32 Hz) temporal frequencies. Selective stimulation of open-field cones that are not in shadow, with penumbral cones silenced, also produced the percept, but it was not seen when penumbral and open-field cones were modulated together. This indicates the need for spatial contrast between penumbral and open-field cones to create the Purkinje-tree percept. Our observation provides a new means for studying the response of retinally stabilized images and demonstrates that penumbral cones can support spatial vision. Further, the result illustrates a way in which silent substitution techniques can fail to be silent. We show that inadvertent penumbral cone stimulation can accompany melanopsin-directed modulations that are designed only to silence open-field cones. This in turn can result in visual responses that might be mistaken as melanopsin-driven.

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          Most cited references62

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          Melanopsin and rod-cone photoreceptive systems account for all major accessory visual functions in mice.

          In the mammalian retina, besides the conventional rod-cone system, a melanopsin-associated photoreceptive system exists that conveys photic information for accessory visual functions such as pupillary light reflex and circadian photo-entrainment. On ablation of the melanopsin gene, retinal ganglion cells that normally express melanopsin are no longer intrinsically photosensitive. Furthermore, pupil reflex, light-induced phase delays of the circadian clock and period lengthening of the circadian rhythm in constant light are all partially impaired. Here, we investigated whether additional photoreceptive systems participate in these responses. Using mice lacking rods and cones, we measured the action spectrum for phase-shifting the circadian rhythm of locomotor behaviour. This spectrum matches that for the pupillary light reflex in mice of the same genotype, and that for the intrinsic photosensitivity of the melanopsin-expressing retinal ganglion cells. We have also generated mice lacking melanopsin coupled with disabled rod and cone phototransduction mechanisms. These animals have an intact retina but fail to show any significant pupil reflex, to entrain to light/dark cycles, and to show any masking response to light. Thus, the rod-cone and melanopsin systems together seem to provide all of the photic input for these accessory visual functions.
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            Diminished pupillary light reflex at high irradiances in melanopsin-knockout mice.

            In the mammalian retina, a small subset of retinal ganglion cells (RGCs) are intrinsically photosensitive, express the opsin-like protein melanopsin, and project to brain nuclei involved in non-image-forming visual functions such as pupillary light reflex and circadian photoentrainment. We report that in mice with the melanopsin gene ablated, RGCs retrograde-labeled from the suprachiasmatic nuclei were no longer intrinsically photosensitive, although their number, morphology, and projections were unchanged. These animals showed a pupillary light reflex indistinguishable from that of the wild type at low irradiances, but at high irradiances the reflex was incomplete, a pattern that suggests that the melanopsin-associated system and the classical rod/cone system are complementary in function.
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              A novel human opsin in the inner retina.

              Here we report the identification of a novel human opsin, melanopsin, that is expressed in cells of the mammalian inner retina. The human melanopsin gene consists of 10 exons and is mapped to chromosome 10q22. This chromosomal localization and gene structure differs significantly from that of other human opsins that typically have four to seven exons. A survey of 26 anatomical sites indicates that, in humans, melanopsin is expressed only in the eye. In situ hybridization histochemistry shows that melanopsin expression is restricted to cells within the ganglion and amacrine cell layers of the primate and murine retinas. Notably, expression is not observed in retinal photoreceptor cells, the opsin-containing cells of the outer retina that initiate vision. The unique inner retinal localization of melanopsin suggests that it is not involved in image formation but rather may mediate nonvisual photoreceptive tasks, such as the regulation of circadian rhythms and the acute suppression of pineal melatonin. The anatomical distribution of melanopsin-positive retinal cells is similar to the pattern of cells known to project from the retina to the suprachiasmatic nuclei of the hypothalamus, a primary circadian pacemaker.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                21 April 2015
                2015
                : 10
                : 4
                : e0124328
                Affiliations
                [1 ]Department of Psychology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America
                [2 ]Department of Neurology, University of Pennsylvania, Philadelphia, Pennsylvania, United States of America
                University College London, UNITED KINGDOM
                Author notes

                Competing Interests: The authors declare competing financial interests. A provisional patent disclosure on the robust photoreceptor isolation method and its applications has been filed by the University of Pennsylvania with GKA, DHB, and MS as inventors (“ROBUST TARGETING OF PHOTOSENSITIVE MOLECULES / Serial No. 62/049,242”, September 11, 2014). This does not alter the authors’ adherence to all the PLoS ONE policies on sharing data and materials.

                Conceived and designed the experiments: MS GKA DHB. Performed the experiments: MS GKA DHB. Analyzed the data: MS GKA DHB. Wrote the paper: MS GKA DHB.

                Article
                PONE-D-14-56273
                10.1371/journal.pone.0124328
                4405364
                25897842
                869c6cf7-0490-472e-a07c-dd8f60d496a2
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 15 December 2014
                : 3 March 2015
                Page count
                Figures: 6, Tables: 1, Pages: 22
                Funding
                This work was supported by National Institutes of Health Grant 1 R01 EY020516 (GKA), R01 EY10016 (DHB), P30 EY001583 (Core Grant for Vision Research) and Deutscher Akademischer Austauschdienst (MS). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Custom metadata
                The trial-by-trial data for the rating experiment as well as the spectra of the modulations used are provided at http://dx.doi.org/10.6084/m9.figshare.1327815. The data set also includes the data for the supplemental rating experiment and the spectra of the modulation presented to our naïve subject. We have made our software available under an open-source license (Silent Substitution Toolbox; https://github.com/spitschan/SilentSubstitutionToolbox).

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