Abstract
Congenital cataracts are associated with gene mutations, yet the underlying mechanism remains largely unknown. Here we reported an embryonic chick lens model that closely recapitulates the process of cataract formation. We adopted dominant-negative site mutations that cause congenital cataracts, connexin, Cx50E48K, aquaporin 0, AQP0R33C, αA-crystallin, CRYAA R12C and R54C. The recombinant retroviruses containing these mutants were microinjected into the occlusive lumen of chick lenses at early embryonic development. Cx50E48K expression developed cataracts associated with disorganized nuclei and enlarged extracellular spaces. Expression of AQP0R33C resulted in cortical cataracts, enlarged extracellular spaces and distorted fiber cell organization. αA crystallin mutations distorted lens light transmission and increased crystalline protein aggregation. Together, retroviral expression of congenital mutant genes in embryonic chick lenses closely mimics characteristics of human congenital cataracts. This model will provide an effective, reliable in vivo system to investigate the development and underlying mechanism of cataracts and other genetic diseases.
Highlights
Congenital cataracts are associated with gene mutations, yet the underlying mechanism remains largely unknown
We investigated whether the expression of cataract-causing mutants in this embryonic chick system could recapitulate the characteristics of human congenital cataracts
Since αA-crystallin (CRYAA) mutations are reported to cause protein aggregation in congenital cataracts[3,23], we investigated the extent of protein aggregation in the E20 chick lens using sucrose gradient sedimentation analysis. αA-crystallin in E20 embryonic chick lenses concentrated in a major protein peak with sedimentation coefficient at 12.5S (Fig. 8A and Supplementary Fig. 4, upper panel)
Summary
Congenital cataracts are associated with gene mutations, yet the underlying mechanism remains largely unknown. Retroviral expression of congenital mutant genes in embryonic chick lenses closely mimics characteristics of human congenital cataracts. This model will provide an effective, reliable in vivo system to investigate the development and underlying mechanism of cataracts and other genetic diseases. There are large numbers of mutated genes to be investigated, far, genetic knockout or knockin mouse models are the only in vivo models available to study congenital cataracts The generation of these animal models is known to be cost-ineffective and time-consuming. Through recombinant retroviruses, we express exogenous dominant-negative site mutants of several lens proteins, Cx50, aquaporin 0, and αA-crystallin that cause congenital cataracts in the embryonic chick lens. We show that analogous to human congenital cataracts, these mutants develop lens opacity, altered lens morphology and structure, and increased protein aggregation
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