Journal article
Dissection of Human Retina and RPE-Choroid for Proteomic Analysis
Journal of Visualized Experiments, Vol.129, e56203
2017
DOI: 10.3791/56203
PMCID: PMC5755368
PMID: 29155757
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.
Details
- Title: Subtitle
- Dissection of Human Retina and RPE-Choroid for Proteomic Analysis
- Creators
- Thiago Cabral - Department of Ophthalmology, Federal University of EspÍrito Santo (UFES)Marcus A Toral - Medical Scientist Training Program, University of IowaGabriel Velez - Medical Scientist Training Program, University of IowaJames E DiCarlo - Edward S. Harkness Eye Institute, New York-Presbyterian HospitalAnuradha M Gore - Omics Laboratory, Byers Eye Institute, Department of Ophthalmology, Stanford UniversityMaryAnn Mahajan - Omics Laboratory, Byers Eye Institute, Department of Ophthalmology, Stanford UniversityStephen H Tsang - Edward S. Harkness Eye Institute, New York-Presbyterian HospitalAlexander G Bassuk - Department of Neurology, University of IowaVinit B Mahajan - Palo Alto Veterans Administration, Palo Alto, CA
- Resource Type
- Journal article
- Publication Details
- Journal of Visualized Experiments, Vol.129, e56203
- Publisher
- MyJove Corporation
- DOI
- 10.3791/56203
- PMID
- 29155757
- PMCID
- PMC5755368
- ISSN
- 1940-087X
- eISSN
- 1940-087X
- Language
- English
- Date published
- 2017
- Academic Unit
- Neurology; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Neurology (Pediatrics)
- Record Identifier
- 9984070855202771
Metrics
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