Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 14 de 14
Filtrar
1.
PLoS One ; 9(9): e106562, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25191897

RESUMO

We aimed to investigate fractalkine (CX3CL1) protein expression in wild type (wt) retina and its alterations during retinal degeneration in mouse model (rd10) of retinitis pigmentosa. Forms of retinal protein CX3CL1, total protein and mRNA levels of CX3CL1 were analyzed at postnatal days (P) 5, 10, 14, 22, 30, 45, and 60 by Western blotting and real-time PCR. Cellular sources of CX3CL1 were investigated by in situ hybridization histochemistry (ISH) and using transgenic (CX3CL1cherry) mice. The immunoblots revealed that in both, wt and rd10 retinas, a membrane integrated ∼100 kDa CX3CL1 form and a cleaved ∼85 kDa CX3CL1 form were present at P5. At P10, accumulation of another presumably intra-neuronal ∼95 kDa form and a decrease in the ∼85-kDa form were observed. From P14, a ∼95 kDa form became principal in wt retina, while in rd10 retinas a soluble ∼85 kDa form increased at P45 and P60. In comparison, retinas of rd10 mice had significantly lower levels of total CX3CL1 protein (from P10 onwards) and lower CX3CL1 mRNA levels (from P14), even before the onset of primary rod degeneration. ISH and mCherry reporter fluorescence showed neurons in the inner retina layers as principal sites of CX3CL1 synthesis both in wt and rd10 retinas. In conclusion, our results demonstrate that CX3CL1 has a distinctive course of expression and functional regulation in rd10 retina starting at P10. The biological activity of CX3CL1 is regulated by conversion of a membrane integrated to a soluble form during neurogenesis and in response to pathologic changes in the adult retinal milieu. Viable mature neurons in the inner retina likely exhibit a dynamic intracellular storage depot of CX3CL1.


Assuntos
Quimiocina CX3CL1/metabolismo , Retina/metabolismo , Degeneração Retiniana/metabolismo , Animais , Quimiocina CX3CL1/genética , Modelos Animais de Doenças , Feminino , Expressão Gênica , Hibridização In Situ , Masculino , Camundongos , Camundongos Transgênicos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Degeneração Retiniana/genética , Retinose Pigmentar/genética , Retinose Pigmentar/metabolismo
2.
BMC Ophthalmol ; 13: 59, 2013 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-24138794

RESUMO

BACKGROUND: Quantitative evaluation of mosaics of photoreceptors and neurons is essential in studies on development, aging and degeneration of the retina. Manual counting of samples is a time consuming procedure while attempts to automatization are subject to various restrictions from biological and preparation variability leading to both over- and underestimation of cell numbers. Here we present an adaptive algorithm to overcome many of these problems.Digital micrographs were obtained from cone photoreceptor mosaics visualized by anti-opsin immuno-cytochemistry in retinal wholemounts from a variety of mammalian species including primates. Segmentation of photoreceptors (from background, debris, blood vessels, other cell types) was performed by a procedure based on Rudin-Osher-Fatemi total variation (TV) denoising. Once 3 parameters are manually adjusted based on a sample, similarly structured images can be batch processed. The module is implemented in MATLAB and fully documented online. RESULTS: The object recognition procedure was tested on samples with a typical range of signal and background variations. We obtained results with error ratios of less than 10% in 16 of 18 samples and a mean error of less than 6% compared to manual counts. CONCLUSIONS: The presented method provides a traceable module for automated acquisition of retinal cell density data. Remaining errors, including addition of background items, splitting or merging of objects might be further reduced by introduction of additional parameters. The module may be integrated into extended environments with features such as 3D-acquisition and recognition.


Assuntos
Algoritmos , Inteligência Artificial , Interpretação de Imagem Assistida por Computador/métodos , Reconhecimento Automatizado de Padrão/métodos , Células Fotorreceptoras de Vertebrados/citologia , Retinoscopia/métodos , Animais , Mamíferos
3.
Opt Express ; 17(22): 19382-400, 2009 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-19997159

RESUMO

This paper presents a successful combination of ultra-high speed (120,000 depth scans/s), ultra-high resolution optical coherence tomography with adaptive optics and an achromatizing lens for compensation of monochromatic and longitudinal chromatic ocular aberrations, respectively, allowing for non-invasive volumetric imaging in normal and pathologic human retinas at cellular resolution. The capability of this imaging system is demonstrated here through preliminary studies by probing cellular intraretinal structures that have not been accessible so far with in vivo, non-invasive, label-free imaging techniques, including pigment epithelial cells, micro-vasculature of the choriocapillaris, single nerve fibre bundles and collagenous plates of the lamina cribrosa in the optic nerve head. In addition, the volumetric extent of cone loss in two colour-blinds could be quantified for the first time. This novel technique provides opportunities to enhance the understanding of retinal pathogenesis and early diagnosis of retinal diseases.


Assuntos
Aumento da Imagem/instrumentação , Lentes , Retina/patologia , Doenças Retinianas/patologia , Retinoscópios , Tomografia de Coerência Óptica/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos
4.
Vis Neurosci ; 26(2): 167-75, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19250601

RESUMO

The presence, density distribution, and mosaic regularity of cone types were studied in the retina of the diurnal agouti, Dasyprocta aguti. Longwave-sensitive (L-) and shortwave-sensitive (S-) cones were detected by antibodies against the respective cone opsins. L- and S-cones were found to represent around 90 and 10% of the cone population, respectively. There was no evidence for L- and S-opsin coexpression in agouti cones. L-cone densities were highest, up to 14,000/mm2, along a horizontal visual streak located about 2-3 mm dorsal to the optic nerve, and the L-cone distribution showed a dorsoventral asymmetry with higher densities in ventral (about 10,000/mm2) than in dorsal (about 4000/mm2) retinal regions. This L-cone topography parallels the agouti's ganglion cell topography. S-cones had a peak density of 1500-2000/mm2 in the central retinal region but did not form a visual streak. Their distribution also showed a dorsoventral asymmetry with densities around 600/mm2 in dorsal and around 1000/mm2 in ventral retinal regions. The patterning of cone arrays was assessed by the density recovery profile analysis. At all eccentricities evaluated, the S-cone mosaic less efficiently packed than the L-cone mosaic. Rod densities ranged from 47,000/mm2 in peripheral to 64,000/mm2 in central retina, and rod:cone ratios were 4:1-9:1. The comparatively low rod density and high cone proportion appear well adapted to the diurnal lifestyle of the agouti.


Assuntos
Células Fotorreceptoras Retinianas Cones/citologia , Animais , Contagem de Células , Visão de Cores , Opsinas dos Cones/biossíntese , Opsinas dos Cones/ultraestrutura , Imuno-Histoquímica , Células Fotorreceptoras Retinianas Cones/metabolismo , Células Ganglionares da Retina/citologia , Células Fotorreceptoras Retinianas Bastonetes/citologia , Células Fotorreceptoras Retinianas Bastonetes/metabolismo , Opsinas de Bastonetes/biossíntese , Opsinas de Bastonetes/ultraestrutura , Roedores
5.
Opt Express ; 16(15): 11083-94, 2008 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-18648422

RESUMO

Cellular in vivo visualization of the three dimensional architecture of individual human foveal cone photoreceptors is demonstrated by combining ultrahigh resolution optical coherence tomography and a novel adaptive optics modality. Isotropic resolution in the order of 2-3 microm, estimated from comparison with histology, is accomplished by employing an ultrabroad bandwidth Titanium:sapphire laser with 140 nm bandwidth and previous correction of chromatic and monochromatic ocular aberrations. The latter, referred to as pancorrection, is enabled by the simultaneous use of a specially designed lens and an electromagnetically driven deformable mirror with unprecedented stroke for correcting chromatic and monochromatic aberrations, respectively. The increase in imaging resolution allows for resolving structural details of distal elements of individual foveal cones: inner segment zones--myoids and ellipsoids--are differentiated from outer segments protruding into pigment epithelial processes in the retina. The presented technique has the potential to unveil photoreceptor development and pathogenesis as well as improved therapy monitoring of numerous retinal diseases.


Assuntos
Aumento da Imagem/métodos , Interpretação de Imagem Assistida por Computador/métodos , Lasers , Retina/citologia , Retinoscopia/métodos , Tomografia de Coerência Óptica/métodos , Humanos , Sensibilidade e Especificidade
6.
J Vis ; 8(4): 23.1-12, 2008 Apr 23.
Artigo em Inglês | MEDLINE | ID: mdl-18484862

RESUMO

We have examined the presence, the distribution, and the opsin identity of photoreceptor types in the retina of the European mole, Talpa europaea, a subterranean insectivore with regressed morphology of the visual system. Cones and rods were identified using opsin antisera, and their topographies determined from flat-mounted retinas. The retina (total area 0.75 mm(2)) contains about 100,000 photoreceptors, 10-12% of which are cones. Rod density is low (theoretical maximum 127,000 mm(-2)). Cone density peaks in central retina (17,750 mm(-2)). Similar to most mammals, two cone opsins, shortwave-sensitive (S) and middle-to-long-wave-sensitive (M), are present. Cone distribution shows a dorsoventral gradient with higher S cone numbers in ventral retina. Coexpression of S and M opsin occurs in more than 30% of the cones. Partial sequencing of the S opsin gene strongly supports UV sensitivity of the mole S cone photopigment. Amino acids that spectrally tune the S opsin are identical in T. europaea and in mammals with known UV cone photosensitivity. The lens transmits light down to 300 nm. Together, our data suggest that photopic vision and UV sensitivity of a cone pigment play a functional role in the European mole.


Assuntos
Toupeiras/fisiologia , Células Fotorreceptoras de Invertebrados/fisiologia , Raios Ultravioleta , Percepção Visual/fisiologia , Animais , Fotomicrografia , Células Fotorreceptoras de Invertebrados/ultraestrutura , Opsinas de Bastonetes/biossíntese , Opsinas de Bastonetes/efeitos da radiação
7.
J Opt Soc Am A Opt Image Sci Vis ; 25(5): 1181-9, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18451927

RESUMO

An algorithm is presented for processing and analysis of differential interference contrast (DIC) microscopy images of the fovea to study the cone mosaic. The algorithm automatically locates the cones and their boundaries in such images and is assessed by comparison with results from manual analysis. Additional algorithms are presented that analyze the cone positions to extract information on cone neighbor relationships as well as the short-range order and domain structure of the mosaic. The methods are applied to DIC images of the human fovea.


Assuntos
Inteligência Artificial , Fóvea Central/citologia , Interpretação de Imagem Assistida por Computador/métodos , Microscopia de Interferência/métodos , Oftalmoscopia/métodos , Reconhecimento Automatizado de Padrão/métodos , Células Fotorreceptoras Retinianas Cones/citologia , Algoritmos , Humanos , Aumento da Imagem/métodos , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
8.
J Comp Neurol ; 507(3): 1336-50, 2008 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-18189305

RESUMO

This study addresses the correlation of retinal topography with factors such as the visual environment, life style, and behavior for a major mammalian group, the artiodactyls. To provide a broader basis for semiquantitative comparison, short-wavelength-sensitive (S)- and middle-to-long-wavelength-sensitive (M)-opsin cone receptor populations from 25 species from five artiodactyl families and of the African elephant were labeled and sampled. The resulting topographic maps were analyzed with respect to the position and extension of high-density regions. For better parameter differentiation, systematic relationships were statistically normalized. In all species examined, two classes of cones have been detected. In most species, the S-cone maxima were located in the temporodorsal retina, but there are exceptions such as the roe deer with accumulation in the ventral retina. For M-cones, as a consequence of their role in terrain/food assessment and predator detection, the standard topography is L-shaped: a horizontal visual streak including a temporal area centralis is extended by a temporal rim. Its extension is correlated with the animal's body height (P = 0.0017): small species (pudu, mouse deer) tend to have a visual streak only, whereas the giraffe shows a complete dorsal arch of elevated densities. Furthermore, a size-independent habitat correlation was revealed for a similar M-cone pattern (P < 0.0001): mountainous species show a striking specialization around the dorsal retina, pointing to the importance of the inferior visual field in precipitous terrain.


Assuntos
Artiodáctilos/anatomia & histologia , Percepção de Cores/fisiologia , Ecossistema , Células Fotorreceptoras Retinianas Cones/citologia , Animais , Biometria , Contagem de Células/métodos , Classificação , Ecologia , Feminino , Masculino , Acuidade Visual/fisiologia
9.
Vis Neurosci ; 23(3-4): 429-35, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-16961976

RESUMO

In mammals, cone photoreceptor subtypes are thought to establish topographies that reflect the species-relevant properties of the visual environment. Middle- to long-wavelength-sensitive (M) cones are the dominant population and in most species they form an area centralis at the visual axis. Short-wavelength-sensitive (S) cone topographies do not always match this pattern. We here correlate the interrelationship of S and M cone topographies in representatives of several mammalian orders with different visual ecology, including man, cheetah, cat, Eurasian lynx, African lion, wild hog, roe deer, and red deer. Retinas were labeled with opsin antisera and S and M cone distributions as well as S/M cone ratios were mapped. We find that species inhabiting open environments show M cone horizontal streaks (cheetah, pig, deer). Species living in structured habitats (tiger, lynx, red deer) have increased S cone densities along the retinal margin. In species with active vision (cheetah, bear, tiger, man), S cone distributions are more likely to follow the centripetal M cone gradients. Small species show a ventral bias of peak S cone density which either matches the peak of M cone density in a temporal area centralis (diurnal sciurid rodents, tree shrews) or not (cat, manul, roe deer). Thus, in addition to habitat structure, physical size and specific lifestyle patterns (e.g. food acquisition) appear to underlie the independent variations of M and S cone topographies.


Assuntos
Mamíferos/classificação , Células Fotorreceptoras Retinianas Cones/metabolismo , Pigmentos da Retina/metabolismo , Animais , Gatos , Humanos , Mamíferos/anatomia & histologia , Oligorribonucleotídeos Antissenso/metabolismo , Células Fotorreceptoras de Vertebrados , Especificidade da Espécie
10.
Vision Res ; 45(28): 3432-44, 2005 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-16249013

RESUMO

A liquid crystal programmable phase modulator (PPM) is used as correcting device in an adaptive optics system for three-dimensional ultrahigh-resolution optical coherence tomography (UHR OCT). The feasibility of the PPM to correct high order aberrations even when using polychromatic light is studied, showing potential for future clinical use. Volumetric UHR OCT of the living retina, obtained with up 25,000A-scans/s and high resolution enables visualization of retinal features that might correspond to groups of terminal bars of photoreceptors at the external limiting membrane.


Assuntos
Imageamento Tridimensional/métodos , Cristais Líquidos , Retina/anatomia & histologia , Tomografia de Coerência Óptica/métodos , Humanos , Imageamento Tridimensional/instrumentação , Óptica e Fotônica , Tomografia de Coerência Óptica/instrumentação
11.
Exp Eye Res ; 78(6): 1117-25, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15109918

RESUMO

Optical coherence tomography (OCT) has become an established diagnostic tool for the clinical assessment of retinal pathology but correlation of acquired signals with retinal substructures has often been ambiguous. In the monkey retina we have now obtained ultrahigh resolution (UHR) OCT images with 1.4 microm axial x 3 microm transverse resolution from perfusion-fixed eye cups of Macaca fascicularis and optimized the identification of retinal anatomy by correction of spatial artefacts in correlated histology. After resin embedding, serial semithin sections were obtained that corresponded to OCT transects. The direct overlay of features identified in histological sections with corresponding OCT locations was limited by non-linear tissue shrinkage due to dehydration and sectioning stress. In the present study, these misalignments were further corrected by using polygonal spline morphing based on corresponding unequivocal landmarks. The geometric normalization then allowed detailed comparison of both profiles including delicate sublayers of photoreceptor inner- and outer segments. Such correlation will facilitate the extraction of structural information from in vivo ultrahigh resolution OCT images in clinical and experimental applications.


Assuntos
Fóvea Central/ultraestrutura , Macaca fascicularis/anatomia & histologia , Animais , Artefatos , Disco Óptico/ultraestrutura , Células Fotorreceptoras de Vertebrados/ultraestrutura , Retina/ultraestrutura , Tomografia de Coerência Óptica/métodos
12.
Invest Ophthalmol Vis Sci ; 44(4): 1696-703, 2003 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-12657611

RESUMO

PURPOSE: To compare ultrahigh-resolution optical coherence tomography (OCT) cross-sectional images of the pig retina with histology, to evaluate the potential of ultrahigh-resolution OCT for enhanced visualization of intra- and subretinal structures. METHODS: Ultrahigh-resolution OCT images were acquired with 1.4- micro m axial x 3- micro m transverse resolution from in vitro posterior eyecup preparations of the domestic pig. Frozen sections were obtained in precise alignment with OCT tomograms, by using major blood vessels as orientation markers and were counterstained with cresyl violet or unstained and examined by differential interference contrast microscopy. Micrographs from histologic sections were linearly scaled to correct for tissue shrinkage and compared with OCT tomograms. RESULTS: In the proximal retina, ultrahigh-resolution OCT signal bands directly corresponded to the main retinal layers. For the wavelength region used ( approximately 800 nm), axodendritic layers (nerve fiber layer, inner and outer plexiform layers) were more reflective than cell body layers (ganglion cell layer, inner nuclear layer, outer nuclear layer). In the distal retina, substructures of the photoreceptor layer such as the interface between inner and outer segments were visualized, and the retinal pigment epithelium, the choriocapillaris, and superficial choroid layers were resolved. In addition, the time sequence of a retinal detachment event was monitored by ultrahigh-resolution OCT. CONCLUSIONS: In vitro ophthalmic ultrahigh-resolution OCT imaging reveals retinal morphology with unprecedented detail. The specific assignment of OCT signal patterns to retinal substructures provides a basis for improved interpretation of in vivo ophthalmic OCT tomograms of high clinical relevance.


Assuntos
Técnicas de Diagnóstico Oftalmológico , Retina/citologia , Animais , Técnicas de Diagnóstico Oftalmológico/instrumentação , Interferometria , Luz , Fibras Nervosas , Neurônios/citologia , Descolamento Retiniano/patologia , Células Ganglionares da Retina/citologia , Suínos , Tomografia/métodos
13.
Brain Res ; 929(1): 139-46, 2002 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-11852040

RESUMO

Previous immunocytochemical, physiological, and molecular studies have reported that the Syrian hamster lacks a shortwave-sensitive (S) cone photopigment but retains circadian responses to ultraviolet (UV) light. Using opsin antibodies and a sensitive detection protocol, we here show that S opsin immunoreactivity colocalizes with M opsin immunoreactivity in a common type of cone photoreceptor. S opsin signal within individual cone outer segments is low and continuously decreases from the ventral to dorsal retina. Only double-labeled cones were found. During development, S opsin expression precedes that of M opsin, but there is no indication of transdifferentiation. Our results imply that in the Syrian hamster low levels of S opsin colocalize with M opsin in a common cone phenotype. We suggest that, similar to other murid rodents, the S pigment absorbs maximally in the UV range, and thus may contribute to mediating the circadian response of the Syrian hamster to UV light.


Assuntos
Ritmo Circadiano/fisiologia , Transdução de Sinal Luminoso/fisiologia , Células Fotorreceptoras Retinianas Cones/metabolismo , Opsinas de Bastonetes/metabolismo , Raios Ultravioleta , Animais , Animais Recém-Nascidos , Contagem de Células , Cricetinae , Expressão Gênica/fisiologia , Imuno-Histoquímica , Mesocricetus , Camundongos , Fenótipo , Isoformas de Proteínas/metabolismo , Células Fotorreceptoras Retinianas Cones/citologia , Células Fotorreceptoras Retinianas Bastonetes/citologia , Células Fotorreceptoras Retinianas Bastonetes/metabolismo
14.
Rev. bras. biol ; 56(supl.1,pt.2): 199-207, dez. 1996. ilus
Artigo em Inglês | LILACS | ID: lil-196342

RESUMO

The topography and spectral characteristics of mammalian photoreceptors correlate with both, the present ecological demands and the evolutionary history. The South American Opossum is a marsupial mammal with unspecialized habitus and crepuscular lifestyle. A sparse population of cones (max. = 3000/mm2) can be differentiated into four subtypes by morphological, topographical and immunocytochemical criteria. In spite of this unusual diversity the cone types can be split into two functional groups: The population of single cones labeled by antibody OS-2 for short wavelenght sensitive pigments was ubiquitous but at very low densities (200/mm2). The single cones labeled by antibody (COS-1) against long wavelength sensitive pigments constitute the dominant population in the area centralis (2300/mm2). These two single cone types correlate with the pair typically present in placental mammals. Discrimination of spatial and color contrast may be provided by this "modern" set. The COS-1 labeled double and single cones bearing an oil droplet, display a different pattern by being restricted to the inferior (non-tapetal) half of the retina (max = 800/mm2). This additional set of cones with oil droplets and long wavelength pigments is a conservative feature of the opossum retina and other marsupials. As an accessory cone system it is possibly providing enhanced sensitivity at mesopic conditions. During the early evolution of nocturnal mammals with its prominent expansion of rod vision these cone types were conserved but then were lost in placental mammals. Thus the unique features of mammalian are the result of two evolutionary steps: first a reduction of cone based vision, followed by a secondary differentiation of photopic vision and behaviour relying on the remaining set of cones.


Assuntos
Animais , Evolução Biológica , Mamíferos/fisiologia , Gambás/fisiologia , Células Fotorreceptoras/fisiologia , Células Fotorreceptoras Retinianas Cones/fisiologia , Retina/fisiologia , Filogenia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...