RESUMO
Purpose: In the eye lens, cytosolic protein concentrations increase progressively from the periphery to the center, contributing to the gradient of refractive index (GRIN). Aquaporins are membrane proteins of lens fiber cells that regulate water transport and adhesion and interact with cytoskeletal proteins. This study investigates how these membrane proteins contribute to proper development of the lens GRIN. Methods: Loss-of-function deletions of aqp0a and/or aqp0b in zebrafish were generated using CRISPR/Cas9 gene editing. Lenses of single aqp0a-/- mutants, single aqp0b-/- mutants, and double aqp0a-/-/aqp0b-/- mutants from larval to elderly adult stages were measured using x-ray Talbot interferometry at SPring8 in Japan. The three-dimensional GRIN profiles in two orthogonal cross-sectional planes of each lens were analyzed and compared with in vivo images and previous results obtained from wild-type lenses. Results: Single aqp0a-/- mutants tended to show asymmetric GRIN profiles, with the central plateau regions shifted anteriorly. Single aqp0b-/- mutants had smooth, symmetric GRIN profiles throughout development until spoke opacities appeared in several extremely old samples. Double aqp0a-/-/aqp0b-/- mutants showed lower magnitude GRIN profiles, as well as dips in the central plateau region. Conclusions: These findings suggest that Aqp0a and Aqp0b have region-specific functions in the lens: Aqp0a is active peripherally, regulating centralization of the plateau region, and this function cannot be compensated for by Aqp0b. In the lens center, either Aqp0a or Aqp0b is required for formation of the plateau region, as well as for the GRIN to reach its maximum magnitude in mature lenses.
Assuntos
Aquaporinas/fisiologia , Catarata/fisiopatologia , Proteínas do Olho/fisiologia , Cristalino/fisiologia , Refração Ocular/fisiologia , Proteínas de Peixe-Zebra/fisiologia , Peixe-Zebra/fisiologia , Animais , Transporte Biológico/fisiologia , Proteína 9 Associada à CRISPR/genética , Catarata/diagnóstico por imagem , Técnicas de Inativação de Genes , Cristalino/diagnóstico por imagem , Síncrotrons , Microtomografia por Raio-XRESUMO
The eyeball is continually subjected to forces that cause alterations to its shape and dimensions, as well as to its optical components. Forces that induce accommodation result in an intentional change in focus; others, such as the effect of intraocular pressure fluctuations, are more subtle. Although the mechanical properties of the eyeball and its components permit mediation of such subtle forces, the concomitant optical changes are not detected by the visual system. Optical self-adjustment is postulated as the mechanism that maintains image quality. The purpose of this study was to investigate how self-adjustment occurs by using an optical model of the eyeball and to test the requisite optical and biometric conditions.
Assuntos
Acomodação Ocular/fisiologia , Olho , Biometria/métodos , Humanos , Pressão Intraocular , Modelos Anatômicos , Visão OcularRESUMO
The purpose of this study was to evaluate the clinical utility of the output parameters of the ocular response analyzer (ORA) and those calculated from the raw ORA in subjects with healthy eyes and those with suspected glaucoma, and in patients with two types of glaucoma. The raw ORA data were analyzed using a custom software that included the Gaussian filtering of applanation curves for three different window sizes. To the best of our knowledge, these findings present a novel means of optimizing the use of measurements from the ORA, which can refine the characteristics of corneal biomechanics, enabling a distinction between the types of glaucoma and leading to an improvement in diagnosing and early detection.