The Biochemical Analysis of Crystallin Proteins in the Adult Zebrafish Lens

Abstract

Zebrafish (Danio rerio) have become important model organisms to increase human understanding of ophthalmological diseases, specifically cataracts. It has been found that there are conserved visual pathways between zebrafish and humans, which allow the study of cataracts in zebrafish to be exceptionally insightful for the mechanisms behind cataract development in humans. The purpose of this study was to provide a biochemical comparison of the soluble and insoluble proteins in cataract and healthy lenses from adult zebrafish. In order to induce cataracts, 3% hydrogen peroxide was injected into lenses of adult zebrafish and the lens proteins were separated into their soluble and insoluble protein fractions through centrifugation. The concentration of the soluble and insoluble proteins was quantified and the lens proteins were further characterized using SDS-PAGE and Western blot analysis to detect crystallins, the predominant lens protein. Lenses that had induced cataracts had an average soluble protein concentration of 0.933ug/uL, while the average insoluble protein concentration was found to be 0.453ug/uL. Normal, healthy lenses were found to have a higher concentration of soluble proteins and a lower concentration of insoluble proteins. Although the results from the SDS-PAGE and Western blot analysis contained some inconsistencies, this research has provided valuable insight into how the Western blot technique can be improved to better detect for crystallin proteins in the future.

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