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Dissection of Human Vitreous Body Elements for Proteomic Analysis
Journal article   Open access   Peer reviewed

Dissection of Human Vitreous Body Elements for Proteomic Analysis

Jessica M. Skeie and Vinit B. Mahajan
Journal of Visualized Experiments, Vol.47, e2455
01/23/2011
DOI: 10.3791/2455
PMCID: PMC3182653
PMID: 21304469
url
https://doi.org/10.3791/2455View
Published (Version of record) Open Access

Abstract

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.
 Medicine

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