Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

Author Cabral, Thiago Autor UNIFESP Google Scholar
Toral, Marcus A. Google Scholar
Velez, Gabriel Google Scholar
DiCarlo, James E. Google Scholar
Gore, Anuradha M. Google Scholar
Mahajan, MaryAnn Google Scholar
Tsang, Stephen H. Google Scholar
Bassuk, Alexander G. Google Scholar
Mahajan, Vinit B. Google Scholar
Abstract The human retina is composed of the sensory neuroretina and the underlying retinal pigmented epithelium (RPE), which is firmly complexed to the vascular choroid layer. Different regions of the retina are anatomically and molecularly distinct, facilitating unique functions and demonstrating differential susceptibility to disease. Proteomic analysis of each of these regions and layers can provide vital insights into the molecular process of many diseases, including Age-Related Macular Degeneration (AMD), diabetes mellitus, and glaucoma. However, separation of retinal regions and layers is essential before quantitative proteomic analysis can be accomplished. Here, we describe a method for dissection and collection of the foveal, macular, and peripheral retinal regions and underlying RPE-choroid complex, involving regional punch biopsies and manual removal of tissue layers from a human eye. One-dimensional SDS-PAGE as well as downstream proteomic analysis, such as liquid chromatography-tandem mass spectrometry (LC-MS/MS), can be used to identify proteins in each dissected retinal layer, revealing molecular biomarkers for retinal disease.
Keywords Human
retina
dissection
retinal pigmented epithelium
choroid
proteomics
xmlui.dri2xhtml.METS-1.0.item-coverage Cambridge
Language English
Sponsor NIH
Doris Duke Charitable Foundation
Research to Prevent Blindness (RPB), New York, NY
Grant number NIH: K08EY020530
NIH: R01EY024665
NIH: R01EY025225
NIH: R01EY024698
NIH: R21AG050437
NIH: T32GM007337
Doris Duke Charitable Foundation: 2013103
Date 2017
Published in Jove-Journal Of Visualized Experiments. Cambridge, v. , n. 129, p. -, 2017.
ISSN 1940-087X (Sherpa/Romeo, impact factor)
Publisher Journal Of Visualized Experiments
Extent -
Origin http://dx.doi.org/10.3791/56203
Access rights Closed access
Type Article
Web of Science ID WOS:000417688700031
URI https://repositorio.unifesp.br/handle/11600/58226

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