RESUMO
An arginine to cysteine substitution at amino acid position 203 (C203R) is the most common missense mutation in human cone opsin. Linked to color blindness and blue cone monochromacy (BCM), C203 is involved in a crucial disulfide bond required for proper folding. It has previously been postulated that expression of mutant C203R cone opsin exerts a toxic effect on cone photoreceptors, similar to some well-characterized missense mutations in rhodopsin that lead to protein misfolding. In this study, we generated and characterized a BCM mouse model carrying the equivalent C203R mutation (Opn1mwC198R Opn1sw-/-) to investigate the disease mechanism and develop a gene therapy approach for this disorder. Untreated Opn1mwC198R Opn1sw-/- cones phenocopied affected cones in human patients with the equivalent mutation, exhibiting shortened or absent cone outer segments and loss of function. We determined that gene augmentation targeting cones specifically yielded robust rescue of cone function and structure when Opn1mwC198R Opn1sw-/- mice were treated at early ages. Importantly, treated cones displayed elaborated outer segments and replenished expression of crucial cone phototransduction proteins. Interestingly, we were unable to detect OPN1MWC198R mutant opsin at any age. We believe this is the first proof-of-concept study exploring the efficacy of gene therapy in BCM associated with a C203R mutation.
Assuntos
Defeitos da Visão Cromática , Opsinas dos Cones , Células Fotorreceptoras Retinianas Cones , Humanos , Animais , Camundongos , Células Fotorreceptoras Retinianas Cones/metabolismo , Mutação de Sentido Incorreto , Opsinas dos Cones/genética , Opsinas dos Cones/metabolismo , Rodopsina/genéticaRESUMO
The study explored the potential of an animal opsin nonselectively expressed in various neuronal elements of the degenerative retina to restore the impaired visual function. A knockout murine model of inherited retinal dystrophy was used. Mice were injected intravitreally with either a virus carrying the gene of short-wavelength cone opsin associated with a reporter fluorescent protein or a control virus carrying the sequence of a modified fluorescent protein with enhanced membrane tropism. Viral transduction induced pronounced opsin expression in ganglion, bipolar, and horizontal retinal neurons. Behavioral testing included the visually guided task in the trapezoid Morris water maze and showed a partial recovery of the learning ability in the mice whose retinas had been transduced with cone opsin.
Assuntos
Opsinas dos Cones , Degeneração Retiniana , Camundongos , Animais , Opsinas dos Cones/genética , Opsinas dos Cones/metabolismo , Degeneração Retiniana/genética , Degeneração Retiniana/metabolismo , Células Fotorreceptoras Retinianas Cones/metabolismo , Retina , Opsinas/metabolismo , Camundongos KnockoutRESUMO
Fish often change their habitat and trophic preferences during development. Dramatic functional differences between embryos, larvae, juveniles and adults also concern sensory systems, including vision. Here, we focus on the photoreceptors (rod and cone cells) in the retina and their gene expression profiles during development. Using comparative transcriptomics on 63 species, belonging to 23 actinopterygian orders, we report general developmental patterns of opsin expression, mostly suggesting an increased importance of the rod opsin (RH1) gene and the long-wavelength-sensitive cone opsin, and a decreasing importance of the shorter wavelength-sensitive cone opsin throughout development. Furthermore, we investigate in detail ontogenetic changes in 14 selected species (from Polypteriformes, Acipenseriformes, Cypriniformes, Aulopiformes and Cichliformes), and we report examples of expanded cone opsin repertoires, cone opsin switches (mostly within RH2) and increasing rod : cone ratio as evidenced by the opsin and phototransduction cascade genes. Our findings provide molecular support for developmental stage-specific visual palettes of ray-finned fishes and shifts between, which most likely arose in response to ecological, behavioural and physiological factors.
Assuntos
Opsinas dos Cones , Opsinas , Animais , Opsinas/genética , Opsinas de Bastonetes/genética , Opsinas dos Cones/genética , Peixes/genética , Células Fotorreceptoras Retinianas Cones/fisiologia , Expressão GênicaRESUMO
We investigate mutations in trß2, a splice variant of thrb, identifying changes in function, structure, and behavior in larval and adult zebrafish retinas. Two N-terminus CRISPR mutants were identified. The first is a 6BP+1 insertion deletion frameshift resulting in a truncated protein. The second is a 3BP in frame deletion with intact binding domains. ERG recordings of isolated cone signals showed that the 6BP+1 mutants did not respond to red wavelengths of light while the 3BP mutants did respond. 6BP+1 mutants lacked optomotor and optokinetic responses to red/black and green/black contrasts. Both larval and adult 6BP+1 mutants exhibit a loss of red-cone contribution to the ERG and an increase in UV-cone contribution. Transgenic reporters show loss of cone trß2 activation in the 6BP+1 mutant but increase in the density of cones with active blue, green, and UV opsin genes. Antibody reactivity for red-cone LWS1 and LWS2 opsin was absent in the 6BP+1 mutant, as was reactivity for arrestin3a. Our results confirm a critical role for trß2 in long-wavelength cone development.
Assuntos
Visão de Cores/genética , Regulação da Expressão Gênica no Desenvolvimento , Genes erbA/genética , Retina/crescimento & desenvolvimento , Receptores beta dos Hormônios Tireóideos/genética , Animais , Animais Geneticamente Modificados , Diferenciação Celular/genética , Opsinas dos Cones/genética , Opsinas dos Cones/metabolismo , Mutação da Fase de Leitura , Mutação INDEL , Larva , Modelos Animais , Células Fotorreceptoras de Invertebrados/patologia , Retina/citologia , Retina/patologia , Deleção de Sequência , Transativadores/genética , Transativadores/metabolismo , Peixe-Zebra/genética , Peixe-Zebra/crescimento & desenvolvimento , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismoRESUMO
Inherited and age-related retinal degenerative diseases cause progressive loss of rod and cone photoreceptors, leading to blindness, but spare downstream retinal neurons, which can be targeted for optogenetic therapy. However, optogenetic approaches have been limited by either low light sensitivity or slow kinetics, and lack adaptation to changes in ambient light, and not been shown to restore object vision. We find that the vertebrate medium wavelength cone opsin (MW-opsin) overcomes these limitations and supports vision in dim light. MW-opsin enables an otherwise blind retinitis pigmenotosa mouse to discriminate temporal and spatial light patterns displayed on a standard LCD computer tablet, displays adaption to changes in ambient light, and restores open-field novel object exploration under incidental room light. By contrast, rhodopsin, which is similar in sensitivity but slower in light response and has greater rundown, fails these tests. Thus, MW-opsin provides the speed, sensitivity and adaptation needed to restore patterned vision.
Assuntos
Cegueira/prevenção & controle , Opsinas dos Cones/genética , Terapia Genética/métodos , Optogenética/métodos , Degeneração Retiniana/terapia , Animais , Cegueira/etiologia , Linhagem Celular , Dependovirus/genética , Modelos Animais de Doenças , Vetores Genéticos/administração & dosagem , Vetores Genéticos/genética , Humanos , Injeções Intravítreas , Queratinócitos , Camundongos , Camundongos Endogâmicos C57BL , Técnicas de Patch-Clamp , Retina/patologia , Células Fotorreceptoras Retinianas Cones/patologia , Degeneração Retiniana/complicações , Degeneração Retiniana/patologia , Rodopsina/genética , Resultado do TratamentoRESUMO
Optogenetics and chemogenetics provide the ability to modulate neurons in a type- and region-specific manner. These powerful techniques are useful to test hypotheses regarding the neural circuit mechanisms of general anesthetic end points such as hypnosis and analgesia. With both techniques, a genetic strategy is used to target expression of light-sensitive ion channels (opsins) or designer receptors exclusively activated by designer drugs in specific neurons. Optogenetics provides precise temporal control of neuronal firing with light pulses, whereas chemogenetics provides the ability to modulate neuronal firing for several hours with the single administration of a designer drug. This chapter provides an overview of neuronal targeting and experimental strategies and highlights the important advantages and disadvantages of each technique.
Assuntos
Anestésicos Gerais/farmacologia , Encéfalo/efeitos dos fármacos , Drogas Desenhadas/farmacologia , Hipnóticos e Sedativos/farmacologia , Neurônios/efeitos dos fármacos , Optogenética/métodos , Anestésicos Gerais/síntese química , Animais , Antipsicóticos/farmacologia , Encéfalo/fisiologia , Clozapina/análogos & derivados , Clozapina/farmacologia , Opsinas dos Cones/genética , Opsinas dos Cones/metabolismo , Dependovirus/genética , Dependovirus/metabolismo , Drogas Desenhadas/síntese química , Diterpenos/farmacologia , Diterpenos Clerodânicos , Eletroencefalografia , Expressão Gênica , Humanos , Hipnose Anestésica/métodos , Hipnóticos e Sedativos/síntese química , Camundongos , Neurônios/citologia , Neurônios/fisiologia , Optogenética/instrumentação , Ratos , Receptor Muscarínico M3/genética , Receptor Muscarínico M3/metabolismo , Receptores Artificiais/genética , Receptores Artificiais/metabolismo , Receptores Opioides kappa/genética , Receptores Opioides kappa/metabolismo , Reflexo de Endireitamento/efeitos dos fármacos , Reflexo de Endireitamento/fisiologia , Técnicas EstereotáxicasRESUMO
Repeated adaptation to a new environment often leads to convergent phenotypic changes whose underlying genetic mechanisms are rarely known. Here, we study adaptation of color vision in threespine stickleback during the repeated postglacial colonization of clearwater and blackwater lakes in the Haida Gwaii archipelago. We use whole genomes from 16 clearwater and 12 blackwater populations, and a selection experiment, in which stickleback were transplanted from a blackwater lake into an uninhabited clearwater pond and resampled after 19 y to test for selection on cone opsin genes. Patterns of haplotype homozygosity, genetic diversity, site frequency spectra, and allele-frequency change support a selective sweep centered on the adjacent blue- and red-light sensitive opsins SWS2 and LWS. The haplotype under selection carries seven amino acid changes in SWS2, including two changes known to cause a red-shift in light absorption, and is favored in blackwater lakes but disfavored in the clearwater habitat of the transplant population. Remarkably, the same red-shifting amino acid changes occurred after the duplication of SWS2 198 million years ago, in the ancestor of most spiny-rayed fish. Two distantly related fish species, bluefin killifish and black bream, express these old paralogs divergently in black- and clearwater habitats, while sticklebacks lost one paralog. Our study thus shows that convergent adaptation to the same environment can involve the same genetic changes on very different evolutionary time scales by reevolving lost mutations and reusing them repeatedly from standing genetic variation.
Assuntos
Adaptação Ocular/efeitos da radiação , Opsinas dos Cones/genética , Evolução Molecular , Proteínas de Peixes/genética , Variação Genética/efeitos da radiação , Smegmamorpha/fisiologia , Luz Solar , Alelos , Substituição de Aminoácidos , Animais , Colúmbia Britânica , Visão de Cores/efeitos da radiação , Opsinas dos Cones/metabolismo , Proteínas de Peixes/metabolismo , Duplicação Gênica , Frequência do Gene , Interação Gene-Ambiente , Genômica/métodos , Ilhas , Lagos , Oceano Pacífico , Filogenia , Smegmamorpha/genética , Smegmamorpha/crescimento & desenvolvimento , Especificidade da EspécieRESUMO
Peripherin-2 is a glycomembrane protein exclusively expressed in the light-sensing compartments of rod and cone photoreceptors designated as outer segments (OS). Mutations in peripherin-2 are associated with degenerative retinal diseases either affecting rod or cone photoreceptors. While peripherin-2 has been extensively studied in rods, there is only little information on its supramolecular organization and function in cones. Recently, we have demonstrated that peripherin-2 interacts with the light detector rhodopsin in OS of rods. It remains unclear, however, if peripherin-2 also binds to cone opsins. Here, using a combination of co-immunoprecipitation analyses, transmission electron microscopy (TEM)-based immunolabeling experiments, and quantitative fluorescence resonance energy transfer (FRET) measurements in cone OS of wild type mice, we demonstrate that peripherin-2 binds to both, S-opsin and M-opsin. However, FRET-based quantification of the respective interactions indicated significantly less stringent binding of peripherin-2 to S-opsin compared to its interaction with M-opsin. Subsequent TEM-studies also showed less co-localization of peripherin-2 and S-opsin in cone OS compared to peripherin-2 and M-opsin. Furthermore, quantitative FRET analysis in acutely isolated cone OS revealed that the cone degeneration-causing V268I mutation in peripherin-2 selectively reduced binding to M-opsin without affecting the peripherin-2 interaction to S-opsin or rhodopsin. The differential binding of peripherin-2 to cone opsins and the mutant-specific interference with the peripherin-2/M-opsin binding points to a novel role of peripherin-2 in cones and might contribute to understanding the differential penetrance of certain peripherin-2 mutations in rods and cones. Finally, our results provide a proof-of-principle for quantitative FRET measurements of protein-protein interactions in cone OS.
Assuntos
Antígenos de Neoplasias/metabolismo , Opsinas dos Cones/metabolismo , Células Fotorreceptoras Retinianas Cones/metabolismo , Degeneração Retiniana/genética , Animais , Antígenos de Neoplasias/genética , Opsinas dos Cones/genética , Transferência Ressonante de Energia de Fluorescência , Humanos , Camundongos , Microscopia Eletrônica de Transmissão , Mutação , Ligação Proteica , Retina/metabolismo , Retina/patologia , Células Fotorreceptoras Retinianas Cones/patologia , Degeneração Retiniana/patologia , Rodopsina/genética , Rodopsina/metabolismoRESUMO
In 524 Japanese individuals with deutan colour vision defect, 76 had a normal-order pigment gene array, where the L gene is at the first position and the M gene(s) is located downstream. Of these 76 individuals, 69 had a -71A>C substitution in the M gene without any other mutation. Because the expression of L/M genes is up-regulated by thyroid hormone (T3) in human retinoblastoma WERI cells, we examined the effects of T3 on promoter activity; T3 increased the activity of the -71A promoter 2-fold, but it had no effect on the -71C promoter. Similarly, the -71C promoter was much less activated by T3 than the -71A promoter in HEK293 cells expressing thyroid hormone receptor isoform ß2. Such a weak response of the -71C promoter to T3 may cause a decrease in the number of M cones and/or the density of M pigment during the differentiation of M cones. The average Rayleigh match midpoint was 18.9 ± 4.1 in 162 ordinary deuteranomaly individuals, but was 37.3 ± 9.1 in 63 deuteranomaly individuals with -71C. The -71A>C substitution was found to be specific to eastern Asia. These results suggest that there may be a new subset of deuteranomaly associated with -71C in the Japanese (and probably eastern Asian) population(s).
Assuntos
Defeitos da Visão Cromática/genética , Opsinas dos Cones/genética , Pigmentos da Retina/genética , Defeitos da Visão Cromática/patologia , Opsinas dos Cones/metabolismo , Feminino , Células HEK293 , Humanos , Japão , Masculino , Mutação , Regiões Promotoras Genéticas , Células Fotorreceptoras Retinianas Cones/metabolismo , Células Fotorreceptoras Retinianas Cones/patologia , Pigmentos da Retina/metabolismo , Receptores beta dos Hormônios Tireóideos/genética , Receptores beta dos Hormônios Tireóideos/metabolismo , Tri-Iodotironina/genética , Tri-Iodotironina/metabolismoRESUMO
To have color vision, having at least two cone photopigment types with different spectral sensitivities present in distinct photoreceptors is necessary together with the neural circuitry necessary to extract color information. Visual pigments are highly conserved molecules, but differences can be found among vertebrate groups. Primates have a variety of cone photopigments (i.e., opsins) that are expressed by polymorphic genes. This article examines the diversity of cone photopigments in New World monkeys and their behavioral relevance...
Assuntos
Animais , Opsinas dos Cones/genética , Platirrinos/genética , Visão de Cores/genéticaRESUMO
PURPOSE: The 65 kDa retinal pigment epithelium-specific protein, RPE65, is an essential enzyme for 11-cis-retinal synthesis in the eye. Mutations of the RPE65 gene in humans result in severe vision loss, and Rpe65(-/-) mice show early cone photoreceptor degeneration. We used an explant culture system to evaluate whether posttranslational downregulation of M-opsin protein in Rpe65(-/-) mice is caused by proteolytic degradation. METHODS: The eyes of three-week-old Rpe65(-/-) mice were incubated in culture medium. Western blot analysis was used to evaluate the level of M-opsin protein, and immunofluorescence was used for protein localization. The transcriptional level of M-opsin was evaluated with real-time reverse-transcriptase-PCR. RESULTS: Degradation of the M-opsin protein in Rpe65(-/-) mouse retina was inhibited by the proteasome inhibitor MG-132 but not by the lysosomal inhibitor pepstatin A and E64d. 9-cis-retinal, used as an analog of 11-cis-retinal, increased M-opsin protein but did not increase M-opsin mRNA. Moreover, 9-cis-retinal did not change the transcriptional levels of photoreceptor specific genes. CONCLUSIONS: Our data suggest that M-opsin protein was degraded through a proteasome pathway and that M-opsin degradation was suppressed with 9-cis-retinal treatment in Rpe65(-/-) mice to some extent.
Assuntos
Opsinas dos Cones/metabolismo , Olho/efeitos dos fármacos , Leupeptinas/farmacologia , Inibidores de Proteassoma , cis-trans-Isomerases/genética , Animais , Opsinas dos Cones/genética , Inibidores de Cisteína Proteinase/farmacologia , Diterpenos , Olho/metabolismo , Leucina/análogos & derivados , Leucina/farmacologia , Lisossomos/metabolismo , Camundongos , Camundongos Knockout , Técnicas de Cultura de Órgãos , Pepstatinas/farmacologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteólise/efeitos dos fármacos , Reação em Cadeia da Polimerase em Tempo Real , Retinaldeído/farmacologia , Transcrição Gênica/efeitos dos fármacos , cis-trans-Isomerases/deficiênciaRESUMO
BACKGROUND: Enhanced S-cone syndrome (ESCS) is an autosomal recessive retinal disorder characterized by an increased number of S-cones over L/M cones and rods. Mutations in the NR2E3 gene, encoding a photoreceptor-specific nuclear receptor, are identified in patients with ESCS. The purpose of this study is to report the ophthalmic features of a 25-year-old Portuguese male with a typical ESCS phenotype and a novel homozygous NR2E3 mutation. METHODS: The patient underwent a detailed ophthalmic examination including fundus photography, fluorescein angiography (FAF), fundus autofluorescence imaging (FAI), and spectral domain optical coherence tomography (SD-OCT). Full-field electroretinography (ERG), S-cone ERG, and multifocal ERG were performed. Mutation screening of the NR2E3 gene was performed with polymerase chain reaction amplification and direct sequencing. RESULTS: The patient had poor visual acuity but good color vision. Funduscopy showed degenerative changes from the vascular arcades to the midperipheral retina. The SD-OCT revealed macular schisis and cystoid changes that had no fluorescein leakage. The posterior pole showed diffusely increased autofluorescence compared with eccentric areas in both eyes. International-standard full-field ERG showed the typical pathognomonic changes associated with ESCS and the short-wavelength flash ERG was simplified, delayed, and similar to the standard photopic flash ERG. Multifocal ERG showed widespread delay and reduction. Genetic analysis revealed a novel homozygous mutation (p.C83Y), which resides in the second zinc finger of the DNA-binding domain. CONCLUSIONS: This homozygous mutation is likely to affect binding to target DNA sites, resulting in a non-functional behavior of NR2E3 protein. It is associated with a typical form of ESCS with a nondetectable rod response and reduced/delayed mfERG responses at all eccentricities.
Assuntos
Opsinas dos Cones/genética , Proteínas do Olho/genética , Receptores Nucleares Órfãos/genética , Mutação Puntual , Células Fotorreceptoras Retinianas Cones/fisiologia , Degeneração Retiniana/genética , Dedos de Zinco/genética , Adulto , Análise Mutacional de DNA , Eletrorretinografia , Éxons/genética , Angiofluoresceinografia , Humanos , Masculino , Reação em Cadeia da Polimerase , Degeneração Retiniana/diagnóstico , Degeneração Retiniana/fisiopatologia , Síndrome , Tomografia de Coerência Óptica , Acuidade Visual/fisiologia , Testes de Campo Visual , Campos VisuaisRESUMO
The successful restoration of visual function with recombinant adeno-associated virus (rAAV)-mediated gene replacement therapy in animals and humans with an inherited disease of the retinal pigment epithelium has ushered in a new era of retinal therapeutics. For many retinal disorders, however, targeting of therapeutic vectors to mutant rods and/or cones will be required. In this study, the primary cone photoreceptor disorder achromatopsia served as the ideal translational model to develop gene therapy directed to cone photoreceptors. We demonstrate that rAAV-mediated gene replacement therapy with different forms of the human red cone opsin promoter led to the restoration of cone function and day vision in two canine models of CNGB3 achromatopsia, a neuronal channelopathy that is the most common form of achromatopsia in man. The robustness and stability of the observed treatment effect was mutation independent, but promoter and age dependent. Subretinal administration of rAAV5-hCNGB3 with a long version of the red cone opsin promoter in younger animals led to a stable therapeutic effect for at least 33 months. Our results hold promise for future clinical trials of cone-directed gene therapy in achromatopsia and other cone-specific disorders.
Assuntos
Defeitos da Visão Cromática/terapia , Canais de Cátion Regulados por Nucleotídeos Cíclicos/genética , Terapia Genética , Células Fotorreceptoras Retinianas Cones , Animais , Defeitos da Visão Cromática/genética , Opsinas dos Cones/genética , Dependovirus/genética , Cães , Feminino , Vetores Genéticos , Masculino , Modelos Animais , Regiões Promotoras Genéticas , Células Fotorreceptoras Retinianas Cones/metabolismo , TransgenesRESUMO
PURPOSE: One potential therapy for vision loss from photoreceptor degeneration is cell replacement, but this approach presents a need for photoreceptor cells. This study explores whether the retinal pigment epithelium (RPE) could be a convenient source of developing photoreceptors. METHODS: The RPE of chick embryos was subjected to reprogramming by proneural genes neurogenin (ngn)1 and ngn3. The genes were introduced into the RPE through retrovirus RCAS-mediated transduction, with the virus microinjected into the eye or added to retinal pigment epithelial explant culture. The retinal pigment epithelia were then analyzed for photoreceptor traits. RESULTS: In chick embryos infected with retrovirus RCAS-expressing ngn3 (RCAS-ngn3), the photoreceptor gene visinin (the equivalent of mammalian recoverin) was expressed in cells of the retinal pigment epithelial layer. When isolated and cultured as explants, retinal pigment epithelial tissues from embryos infected with RCAS-ngn3 or RCAS-ngn1 gave rise to layers of visinin-positive cells. These reprogrammed cells expressed genes of phototransduction and synapses, such as red opsin, the alpha-subunit of cone transducin, SNAP-25, and PSD-95. Reprogramming occurred with retinal pigment epithelial explants derived from virally infected embryos and with retinal pigment epithelial explants derived from normal embryos, with the recombinant viruses added at the onset of the explant culture. In addition, reprogramming took place in retinal pigment epithelial explants from both young and old embryos, from embryonic day (E)6 to E18, when the visual system becomes functional in the chick. CONCLUSIONS: The results support the prospect of exploring the RPE as a convenient source of developing photoreceptors for in situ cell replacement.
Assuntos
Proteínas Aviárias/metabolismo , Diferenciação Celular , Reprogramação Celular , Proteínas do Tecido Nervoso/fisiologia , Células Fotorreceptoras de Vertebrados/citologia , Neurônios Retinianos/citologia , Epitélio Pigmentado da Retina/citologia , Animais , Fatores de Transcrição Hélice-Alça-Hélice Básicos/fisiologia , Células Cultivadas , Embrião de Galinha , Opsinas dos Cones/genética , Opsinas dos Cones/metabolismo , Técnica Indireta de Fluorescência para Anticorpo , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Vetores Genéticos , Hibridização In Situ , Peptídeos e Proteínas de Sinalização Intracelular/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Células Fotorreceptoras de Vertebrados/metabolismo , Neurônios Retinianos/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Retroviridae/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteína 25 Associada a Sinaptossoma/genética , Proteína 25 Associada a Sinaptossoma/metabolismo , Transducina/genética , Transducina/metabolismo , TransfecçãoRESUMO
Missense mutations in the cone opsins have been identified as a relatively common cause of red/green color vision defects, with the most frequent mutation being the substitution of arginine for cysteine at position 203 (C203R). When the corresponding cysteine is mutated in rhodopsin, it disrupts proper folding of the pigment, causing severe, early onset retinitis pigmentosa. While the C203R mutation has been associated with loss of cone function in color vision deficiency, it is not known what happens to cones expressing this mutant opsin. Here, we used high-resolution retinal imaging to examine the cone mosaic in two individuals with genes encoding a middle-wavelength sensitive (M) pigment with the C203R mutation. We found a significant reduction in cone density compared to normal and color-deficient controls, accompanying disruption in the cone mosaic in both individuals, and thinning of the outer nuclear layer. The C203R mosaics were different from that produced by another mutation (LIAVA) previously shown to disrupt the cone mosaic. Comparison of these mosaics provides insight into the timing and degree of cone disruption and has implications for the prospects for restoration of vision loss associated with various cone opsin mutations.
Assuntos
Substituição de Aminoácidos/genética , Opsinas dos Cones/genética , Mosaicismo , Mutação/genética , Adulto , Arginina/genética , Contagem de Células , Cromossomos Humanos X/genética , Visão de Cores/genética , Cisteína/genética , Técnicas de Diagnóstico Oftalmológico , Feminino , Humanos , Masculino , Retina/patologia , Células Fotorreceptoras Retinianas Cones/patologia , Adulto JovemRESUMO
Gulls (Laridae excluding Sternidae) appear to be the only shorebirds (Charadriiformes) that have a short wavelength sensitive type 1 (SWS1) cone pigment opsin tuned to ultraviolet (UV) instead of violet. However, the apparent UV-sensitivity has only been inferred indirectly, via the interpretation that the presence of cysteine at the key amino acid position 90 in the SWS1 opsin confers UV sensitivity. Unless the cornea and the lens efficiently transmit UV to the retina, gulls might in effect be similar to violet-sensitive birds in spectral sensitivity even if they have an ultraviolet sensitive (UVS) SWS1 visual pigment. We report that the spectral transmission of the cornea and lens of great black-backed Larus marinus and herring gulls L. argentatus allow UV-sensitivity, having a lambda(T0.5) value, 344 nm, similar to the ocular media of UV sensitive birds. By molecular sequencing of the second alpha-helical transmembrane region of the SWS1 opsin gene we could also infer that 15 herring gulls and 16 yellow-legged gulls L. michahellis, all base-pair identical, are genetically UV-sensitive.