The vertebrate ??crystallins play an essential role in lens development and the maintenance of lens transparency. Abnormal expression of ??crystallins is also involved in numerous human diseases of the eye, muscular and nervous systems, and has recently been implicated in various cancers. Our goals in this proposal are to leverage the benefits of the zebrafish as a model system to 1) detail the roles of ??crystallins in lens development and 2) identify novel regulatory regions in the mouse and zebrafish 1?? -crystallin promoter. These studies will contribute to our understanding of normal and pathological lens function. We propose testing the hypothesis that the role of ??crystallins in lens development is conserved between zebrafish and mammals by using synthetic anti-sense oligonucleotide molecules called morpholinos to inhibit translation of each zebrafish ??crystallin individually and in combination. This work will provide an efficient model system for analyzing the specifics of ?? crystallin developmental function, and will allow future in vivo studies of genetically modified ??crystallins. The unusual presence of two 1?? -crystallins in zebrafish (one lens specific and one ubiquitously expressed in the body) will allow us to examine how the functions of mammalian 1?-crystallin have become divided between the two zebrafish paralogs. The zebrafish will also be used as a rapid screening system to identify novel regulatory regions controlling 1?? -crystallin's temporal and spatial expression. Regulatory elements are so far only known from a relatively small proximal region of the mammalian 1?? -crystallin promoter. Future studies can utilize this zebrafish model to test the in vivo effects of mutated promoters on ??crystallin expression. The proposed work will be done in an undergraduate biology department, allowing students to gain experience in experimental design, data analysis and cutting edge research techniques used in visual science.

Public Health Relevance

??crystallin proteins are necessary for eye lens development and the maintenance of lens transparency. Mutations in these proteins and abnormal amounts are associated with multiple diseases of the eye and nervous system and found in various cancers. This project will use the zebrafish as a model species for examining the role of ??crystallins in lens development and the mechanisms that control their production.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
2R15EY013535-03
Application #
7780221
Study Section
Anterior Eye Disease Study Section (AED)
Program Officer
Araj, Houmam H
Project Start
2001-08-01
Project End
2014-01-31
Budget Start
2010-02-01
Budget End
2014-01-31
Support Year
3
Fiscal Year
2010
Total Cost
$184,544
Indirect Cost
Name
Ashland University
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
076751825
City
Ashland
State
OH
Country
United States
Zip Code
44805
Posner, Mason; Skiba, Jackie; Brown, Mary et al. (2013) Loss of the small heat shock protein ?A-crystallin does not lead to detectable defects in early zebrafish lens development. Exp Eye Res 116:227-33
Wages, Phillip; Horwitz, Joseph; Ding, Linlin et al. (2013) Changes in zebrafish (Danio rerio) lens crystallin content during development. Mol Vis 19:408-17
Posner, Mason; Kiss, Andor J; Skiba, Jackie et al. (2012) Functional validation of hydrophobic adaptation to physiological temperature in the small heat shock protein ?A-crystallin. PLoS One 7:e34438
Posner, Mason; Hawke, Molly; Lacava, Carrie et al. (2008) A proteome map of the zebrafish (Danio rerio) lens reveals similarities between zebrafish and mammalian crystallin expression. Mol Vis 14:806-14
Smith, Amber A; Wyatt, Keith; Vacha, Jennifer et al. (2006) Gene duplication and separation of functions in alphaB-crystallin from zebrafish (Danio rerio). FEBS J 273:481-90
Dahlman, Jason M; Margot, Kelli L; Ding, Linlin et al. (2005) Zebrafish alpha-crystallins: protein structure and chaperone-like activity compared to their mammalian orthologs. Mol Vis 11:88-96
Runkle, Stephanie; Hill, Julie; Kantorow, Marc et al. (2002) Sequence and spatial expression of zebrafish (Danio rerio) alphaA-crystallin. Mol Vis 8:45-50