Dissection of Human Vitreous Body Elements for Proteomic Analysis
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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.
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Ocular tissues, including the retina, shed intracellular proteins into adjacent intraocular fluid compartments such as vitreous fluid.
“proteomic studies of the eye reveal that ocular tissues, including the retina, shed intracellular proteins into adjacent intraocular fluid compartments (e.g., the vitreous fluid; Fig. 1a)20,26–28.”
From Investigation of Cas9 Antibodies in the Human Eye (2022) · cites this paper as [27]
Vitreous fluid sequesters both intracellular and extracellular proteins shed from the retina and adjacent tissues.
“This tissue makes up 80% of the human eye by volume and sequesters both intracellular and extracellular proteins shed from the retina and adjacent tissues20,26–28.”
From Investigation of Cas9 Antibodies in the Human Eye (2022) · cites this paper as [27]
Vitreous fluid makes up 80% of the human eye by volume.
“This tissue makes up 80% of the human eye by volume and sequesters both intracellular and extracellular proteins shed from the retina and adjacent tissues20,26–28.”
From Investigation of Cas9 Antibodies in the Human Eye (2022) · cites this paper as [27]
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References 6 · Cited by 28
- Authors
- Skeie, J. M., & Mahajan, V. B.
- Affiliations
- University of Iowa (US)
- ORCID
- Mahajan, Vinit B.
- Venue
- Journal of Visualized Experiments
- Publisher
- MyJove Corporation
- Published
- 2011-01-23
- Issue
- 47
- ISSN
- 1940-087X (ISSN-L)
- DOI
- 10.3791/2455
- PMC
- PMC3182653
- OpenAlex
- W2011265611
- Cited by
- 28
- References
- 6
- Citations by year
- 2022: 2 · 2021: 1 · 2020: 2 · 2019: 1 · 2018: 2 · 2017: 3 · 2016: 5 · 2015: 2 · 2014: 4 · 2013: 2 · 2012: 3
- FWCI
- 2.44
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- cc-by-nc-nd
- Locations
- Journal of Visualized Experiments (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- Funders
- National Eye Institute (F32EY022280)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Retinal and Macular Surgery
- Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Keywords
- vitreous base, vitreous cortex, vitreoretinal disease, vitreous proteome, protein compartmentalization, vitreomacular traction, macular hole, retinal detachment
- MeSH terms
- Dissection, Electrophoresis, Polyacrylamide Gel, Humans, Vitreous Body, Proteomics
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