Document Detail

An ENU mutagenesis screen in zebrafish for visual system mutants identifies a novel splice-acceptor site mutation in patched2 that results in Colobomas.
MedLine Citation:
PMID:  23150614     Owner:  NLM     Status:  MEDLINE    
PURPOSE: To identify recessive mutations affecting development and/or maintenance of the zebrafish visual system.
METHODS: A three-generation ENU (N-Nitroso-N-ethylurea)-based forward genetic screen was performed. F3 embryos were screened visually from 1 to 5 days postfertilization (dpf) for ocular abnormalities, and 5 dpf embryos were fixed and processed for cryosectioning, after which eye sections were screened for defects in cellular organization within the retina, lens, and cornea. A combination of PCR and DNA sequencing, in situ hybridization, and pharmacological treatments were used to clone and characterize a coloboma mutant.
RESULTS: A total of 126 F2 families were screened, and, from these, 18 recessive mutations were identified that affected eye development. Phenotypes included lens malformations and cataracts, photoreceptor defects, oculocutaneous albinism, microphthalmia, and colobomas. Analysis of one such coloboma mutant, uta(1), identified a splice-acceptor mutation in the patched2 gene that resulted in an in-frame deletion of 19 amino acids that are predicted to contribute to the first extracellular loop of Patched2. ptch2(uta1) mutants possessed elevated Hedgehog (Hh) pathway activity, and blocking the Hh pathway with cyclopamine prevented colobomas in ptch2(uta1) mutant embryos.
CONCLUSIONS: We have identified 18 recessive mutations affecting development of the zebrafish visual system and we have characterized a novel splice-acceptor site mutation in patched2 that results in enhanced Hh pathway activity and colobomas.
Jiwoon Lee; Ben D Cox; Christina M S Daly; Chanjae Lee; Richard J Nuckels; Rachel K Tittle; Rosa A Uribe; Jeffrey M Gross
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-12-13
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  53     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-14     Completed Date:  2013-02-14     Revised Date:  2013-07-11    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8214-21     Citation Subset:  IM    
Section of Molecular Cell and Developmental Biology, Institute of Cell and Molecular Biology, The University of Texas at Austin, Texas 78712, USA.
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MeSH Terms
Alkylating Agents / toxicity
Amino Acid Sequence
Base Sequence
Coloboma / genetics*
Embryo, Nonmammalian
Ethylnitrosourea / toxicity*
Eye / embryology
Genes, Recessive
Hedgehog Proteins / genetics
In Situ Hybridization
Lens, Crystalline / abnormalities*,  embryology
Membrane Proteins / genetics*
Molecular Sequence Data
Mutagenesis / drug effects
Mutation / genetics*
Polymerase Chain Reaction
RNA Splice Sites / genetics*
Retina / abnormalities*,  embryology
Sequence Analysis, DNA
Veratrum Alkaloids / pharmacology
Zebrafish Proteins / genetics*
Grant Support
Reg. No./Substance:
0/Alkylating Agents; 0/Hedgehog Proteins; 0/Membrane Proteins; 0/RNA Splice Sites; 0/Veratrum Alkaloids; 0/Zebrafish Proteins; 0/ptc2 protein, zebrafish; 759-73-9/Ethylnitrosourea; ZH658AJ192/cyclopamine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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