RESUMO
Leber congenital amaurosis (LCA) is a neurodegenerative disease of photoreceptor cells that causes blindness within the first year of life. It occasionally occurs in syndromic metabolic diseases and plurisystemic ciliopathies. Using exome sequencing in a multiplex family and three simplex case subjects with an atypical association of LCA with early-onset hearing loss, we identified two heterozygous mutations affecting Arg391 in ß-tubulin 4B isotype-encoding (TUBB4B). Inspection of the atomic structure of the microtubule (MT) protofilament reveals that the ß-tubulin Arg391 residue contributes to a binding pocket that interacts with α-tubulin contained in the longitudinally adjacent αß-heterodimer, consistent with a role in maintaining MT stability. Functional analysis in cultured cells overexpressing FLAG-tagged wild-type or mutant TUBB4B as well as in primary skin-derived fibroblasts showed that the mutant TUBB4B is able to fold, form αß-heterodimers, and co-assemble into the endogenous MT lattice. However, the dynamics of growing MTs were consistently altered, showing that the mutations have a significant dampening impact on normal MT growth. Our findings provide a link between sensorineural disease and anomalies in MT behavior and describe a syndromic LCA unrelated to ciliary dysfunction.
Assuntos
Amaurose Congênita de Leber/genética , Microtúbulos/genética , Tubulina (Proteína)/genética , Adulto , Sítios de Ligação/genética , Células Cultivadas , Criança , Análise Mutacional de DNA , Feminino , Humanos , Masculino , Microtúbulos/metabolismo , Pessoa de Meia-Idade , Mutação de Sentido Incorreto/genética , Células Fotorreceptoras/metabolismo , Tubulina (Proteína)/metabolismo , Sequenciamento do ExomaRESUMO
PURPOSE: To report a previously undescribed pattern of crinkled retinal pigment epithelium (RPE), observed in a family of black patients originating from Martinique, an island in the French West Indies. METHODS: Three generations were examined by visual acuity measurement and fundus photography. Autofluorescence photography, fluorescein and indocyanine green angiography, visual field testing, electrophysiology, and spectral domain optical coherence tomography were performed in certain patients. RESULTS: One 86-year-old grandmother, her 7 children, her nephew, and 18 of her 22 grandchildren were examined. Nine patients were affected: five children, one nephew, and three grandchildren. An unrelated patient originating from the same area was also affected. In the third generation, fundus findings were whitish deep lines located in the posterior pole. Optical coherence tomography showed a crinkled pattern of a slightly elevated RPE. In the second generation, a scalloped crinkled RPE was observed in the posterior pole and midperiphery, giving an image of dry desert land in fluorescein and indocyanine green angiography. Optical coherence tomography showed that the RPE formed ripples, giving it a crinkled appearance. Complications were observed in six cases: they included RPE atrophy (one case), subretinal and sub-RPE hemorrhages because of polypoidal choroidal vasculopathy (four cases), and fibrovascular scarring (one case). The grandmother's fundi were characterized by peripheral pigmentary changes, with severe visual loss. CONCLUSION: The observed pattern appeared different from previously described dystrophies and could be referred to as Martinique crinkled retinal pigment epitheliopathy.
Assuntos
Doenças Retinianas , Adulto , Idoso , Idoso de 80 Anos ou mais , Eletrorretinografia , Feminino , Angiofluoresceinografia , Humanos , Verde de Indocianina , Masculino , Martinica , Pessoa de Meia-Idade , Linhagem , Fenótipo , Doenças Retinianas/patologia , Doenças Retinianas/fisiopatologia , Epitélio Pigmentado da Retina/patologia , Tomografia de Coerência Óptica , Acuidade Visual/fisiologia , Campos Visuais/fisiologia , Adulto JovemRESUMO
OBJECTIVE: To obtain images of anterior and posterior segments of the eye using a slit-lamp (SL)/spectral domain (SD) optical coherence tomography (OCT) integrated system designed for the human eye, in the cat, dog, minipig and monkey. ANIMALS STUDIED: One healthy adult monkey, one healthy adult minipig, one healthy adult dog, one healthy adult cat, and three cats and four dogs affected by corneal or retinal diseases. PROCEDURE: A SL SCAN-1 SD-OCT, which is a slit-lamp SL-D7 that contains an integrated OCT module and a fundus viewer, was used to generate OCT images (512-2048), while simultaneously taking 'en-face' slit-lamp images (efSL). OCT images were obtained under sedation or anesthesia. These images were compared to histological retinal sections obtained from a monkey, a minipig, a dog, and a cat. RESULTS: 'en-face' slit-lamp images and OCT images of the ocular tissues were obtained allowing for the identification of different corneal and retinal layers in all animal species. Measurements of the total retinal thickness (TRT) from the inner limiting membrane to the retinal pigment epithelium were performed in various regions throughout the retina. Reduction in TRT was consistent with clinical features of retinal degeneration identified in dogs and cats. CONCLUSION: This noninvasive procedure is useful for both experimental and clinical assessments of ocular tissue damage. Images of anterior and posterior segments are readily obtained under routine clinical conditions. Future studies are warranted to establish normal OCT data in our patients with this new instrument.
Assuntos
Segmento Anterior do Olho/anatomia & histologia , Técnicas de Diagnóstico Oftalmológico/veterinária , Disco Óptico/anatomia & histologia , Segmento Posterior do Olho/anatomia & histologia , Tomografia de Coerência Óptica/veterinária , Animais , Segmento Anterior do Olho/patologia , Técnicas de Diagnóstico Oftalmológico/instrumentação , Oftalmopatias/diagnóstico , Oftalmopatias/patologia , Oftalmopatias/veterinária , Feminino , Masculino , Segmento Posterior do Olho/patologia , Especificidade da Espécie , Tomografia de Coerência Óptica/métodosRESUMO
PURPOSE: To perform cellular-level in vivo imaging of the feline retina using an adaptive optics flood illumination fundus camera (AO FIFC) designed for the human eye. MATERIALS AND METHODS: Cellular-level images were obtained from three eyes of two normal sedated cats. Ocular aberrations were corrected using an AO system based on a 52-acuator electromagnetic deformable mirror and a 1024 lenslet Hartmann-Shack sensor (both Imagine Eyes, Orsay, France). A square 3°×3° area of the ocular fundus was flood-illuminated by a pulsed LED emitting at 850 nm and imaged onto a low-noise, high-resolution CCD camera. The animal's pupils were dilated and the effective pupil size was set to 7.5 mm. Conjunctival atraumatic clips were used to avoid eyeball movements and eyelid closure. The cornea was artificially hydrated throughout the experiments. Each acquisition consisted of 20 consecutive images, out of which 10 were numerically averaged to produce an enhanced final image. RESULTS: The total amount of ocular aberrations was greatly reduced by the AO correction, from 2.4 to 0.21 microns root mean square on average. The resulting images presented white dots distributed at a density similar to that of cone photoreceptors and they allowed us to visualize small blood vessels and nerve fiber bundles at a higher resolution than classically obtained with conventional fundus photography. CONCLUSION: Retinal imaging with cellular resolution was feasible in cats under sedation using an AO FIFC designed for human eyes without any optical modification. The AO FIFC technology could find new applications in clinical, pharmacological, and toxicological investigations.
Assuntos
Gatos/anatomia & histologia , Diagnóstico por Imagem/veterinária , Oftalmoscopia/veterinária , Retina/anatomia & histologia , Animais , Diagnóstico por Imagem/instrumentação , Fenômenos ÓpticosRESUMO
PURPOSE: To analyze high-resolution color photographs of the mouse fundus. METHODS: A contact fundus camera based on topical endoscopy fundus imaging (TEFI) was built. Fundus photographs of C57 and Balb/c mice obtained by TEFI were qualitatively analyzed. RESULTS: High-resolution digital imaging of the fundus, including the ciliary body, was routinely obtained. The reflectance and contrast of retinal vessels varied significantly with the amount of incident and reflected light and, thus, with the degree of fundus pigmentation. The combination of chromatic and spherical aberration favored blue light imaging, in term of both field and contrast. CONCLUSIONS: TEFI is a small, low-cost system that allows high-resolution color fundus imaging and fluorescein angiography in conscious mice. Panretinal imaging is facilitated by the presence of the large rounded lens. TEFI significantly improves the quality of in vivo photography of retina and ciliary process of mice. Resolution is, however, affected by chromatic aberration, and should be improved by monochromatic imaging.
Assuntos
Fundo de Olho , Fotografação/métodos , Retina/anatomia & histologia , Vasos Retinianos/anatomia & histologia , Animais , Angiofluoresceinografia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Fotografação/instrumentaçãoRESUMO
PURPOSE: Angiogenic inhibitors, alone or combined with other therapies, are believed to represent a promising treatment for neovascularization in age-related macular degeneration (wet AMD). They can maintain or improve visual acuity (VA), at least for the first 2years. However, evolution to retinal atrophy cannot be ruled out and it may be useful to assess the effects of antiangiogenic therapy on retinal and choroidal circulation. METHODS: We carried out a pilot study in 15 patients with wet AMD. Time-averaged mean blood flow velocities (BFVs) in the central retinal, temporal posterior ciliary and ophthalmic arteries (CRA, TPCA and OA) were measured by ultrasound imaging before and 4weeks after a single intravitreal injection of 1.25mg bevacizumab in 0.05ml. Patients underwent two ophthalmic examinations, before and 4weeks after injection, including VA measurement and optical coherence tomography (OCT3) examination. RESULTS: In treated eyes, bevacizumab injection was followed by a significant improvement in VA (from 20/125 to 20/80; p=0.0214), and a decrease in mean central macular thickness (from 392±96µm to 271±50µm; p=0.0038). Mean BFV decreased by 10% in the CRA (p=0.0226), 20% in the TPCA (p=0.0026) and 20% in the OA (p=0.0003). No effect was observed in fellow eyes. CONCLUSIONS: Intravitreal bevacizumab acutely improved VA and reduced central macular thickness in wet AMD. Ultrasound imaging revealed that BFVs decreased in all retrobulbar arteries, suggesting that after local diffusion, bevacizumab exerts a short-term regional effect. Bevacizumab might therefore induce hypoperfusion of the whole eye, which may correspond to a vascular side-effect.
Assuntos
Inibidores da Angiogênese/administração & dosagem , Anticorpos Monoclonais/administração & dosagem , Neovascularização de Coroide/tratamento farmacológico , Neovascularização de Coroide/fisiopatologia , Degeneração Macular Exsudativa/tratamento farmacológico , Degeneração Macular Exsudativa/fisiopatologia , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais Humanizados , Bevacizumab , Velocidade do Fluxo Sanguíneo , Pressão Sanguínea , Neovascularização de Coroide/diagnóstico por imagem , Artérias Ciliares/diagnóstico por imagem , Artérias Ciliares/fisiologia , Feminino , Angiofluoresceinografia , Frequência Cardíaca , Humanos , Injeções Intravítreas , Masculino , Pessoa de Meia-Idade , Artéria Oftálmica/diagnóstico por imagem , Artéria Oftálmica/fisiologia , Projetos Piloto , Artéria Retiniana/diagnóstico por imagem , Artéria Retiniana/fisiologia , Tomografia de Coerência Óptica , Ultrassonografia Doppler , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Acuidade Visual/fisiologia , Degeneração Macular Exsudativa/diagnóstico por imagemRESUMO
PURPOSE: The authors recently used topical endoscopy to image the mouse eye fundus. Here, they widened the field of application for this ophthalmologic tool, imaging both the posterior and the anterior eye segments in larger animals commonly encountered in research laboratories and veterinary clinics. METHODS: Pupils were dilated, and local anesthetic and gel were applied to the animal cornea. The endoscopic probe was placed in contact with the cornea of conscious rats, sedated cats and dogs, anesthetized sheep, and nonhuman primates. RESULTS: High-resolution digital images of the eye fundus were obtained in all investigated animals using the endoscopic probe along the eye axis. Arteriovenous filling time was monitored with fluorescein angiography in pigmented rats. The retinal periphery and ciliary bodies could be visualized with the probe placed at an oblique angle. The probe was inclined further to observe the iridocorneal angle such that the pectinate ligaments could be seen at high resolution in cats. The authors used the probe on eyes with retinal detachment, luxation of a cataractous lens, and pigment infiltration in the iridocorneal angle, demonstrating its potential use in eye diseases. CONCLUSIONS: This topical endoscopic technique provides a unique tool for single eye examinations. The authors obtained a circular view of the anterior (iridocorneal angle) and the posterior (fundus) eye segments from all animal species studied. This technique is inexpensive and easy to use. It can be easily moved to the eye of the patient who cannot move to stand in front of classic apparatus, offering new opportunities in ophthalmology.
Assuntos
Corpo Ciliar/patologia , Córnea/patologia , Técnicas de Diagnóstico Oftalmológico/economia , Endoscopia/economia , Oftalmopatias/diagnóstico , Iris/patologia , Retina/patologia , Animais , Callithrix , Gatos , Análise Custo-Benefício , Técnicas de Diagnóstico Oftalmológico/instrumentação , Modelos Animais de Doenças , Cães , Endoscópios/economia , Endoscopia/métodos , Desenho de Equipamento , Haplorrinos , Camundongos , Ratos , Reprodutibilidade dos Testes , OvinosRESUMO
The i-wave, a post b-wave component of the human photopic electroretinogram (ERG), is claimed to originate at the level of the retinal ganglion cells (RGC) or more distally. We investigated whether this wave is a feature common to all species. Photopic ERGs were obtained from the following species: Beagle dog, European cat, New Zealand white rabbit, Göttingen minipig, Cynomolgus monkey, Sprague-Dawley and brown Norway rats, Hartley guinea pig, and CD1 and C57BL6 mice. Results were compared with those obtained from normal human subjects. Except for rats and mice, all species yielded a well-demarcated i-wave, easily identifiable and separated from the a-b-wave complex by approximately 20 ms. Our sample suggests that the i-wave is a feature common to the photopic ERG of most species including humans. In view of its suggested origin, the i-wave would offer a unique opportunity to test, with the flash ERG, the functional integrity of the retinal ganglion cells in animals where use of a pattern stimulus is not always easily obtained.
Assuntos
Retina/fisiologia , Animais , Gatos , Cães , Eletrorretinografia/veterinária , Cobaias , Humanos , Macaca fascicularis , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos , Coelhos , Ratos , Ratos Sprague-Dawley , Especificidade da Espécie , Porco MiniaturaRESUMO
PURPOSE: The purpose of this study was to explore the presence of the filling-in phenomenon in patients with uni- or bilateral central scotoma (CS) resulting from natural history or laser photocoagulation of choroidal neovascularization in age-related macular degeneration (AMD). METHODS: Sixteen consecutive patients with unilateral CS and 14 patients with bilateral CS were assessed (44 eyes) with a scanning laser ophthalmoscope (SLO). Scotoma was delineated by scotometry with a point (1 degree x1 degree) moving radially from the periphery to the center of the lesion. In addition, patients underwent a line test, consisting of a horizontal line moving vertically and a vertical line moving horizontally, from the periphery to the center. The lines were longer than the macular lesion and were projected onto the retina. Patients were asked to indicate when the lines seemed interrupted. The perceptual filling-in phenomenon was considered to be present when limits of the perceived scotoma, determined by the line test, were smaller than those assessed by scotometry. In patients with bilateral CS, the results were analyzed to distinguish the less or more severely affected eye. RESULTS: In all eyes, the limits of the scotoma obtained with the scotometry test corresponded to the anatomic edges of the macular lesion. In patients with bilateral CS, the filling-in phenomenon was observed in 12 out of 14 (85%) less severely affected eyes, but only in one (7%) of their more severely affected eyes. In patients with unilateral CS, the phenomenon was observed in only one out of 16 (6%) eyes. CONCLUSION: These results suggest that the filling-in phenomenon mostly occurs in patients with bilateral central scotoma, and almost always in their less affected eye. Thus, it did usually not occur in an eye if the fellow eye was better.