Document Detail

Gene-specific differential response to anti-apoptotic therapies in zebrafish models of ocular coloboma.
MedLine Citation:
PMID:  21677791     Owner:  NLM     Status:  MEDLINE    
PURPOSE: We recently demonstrated that molecular therapy using aminoglycosides can overcome the underlying genetic defect in two zebrafish models of ocular coloboma and showed abnormal cell death to be a key feature associated with the optic fissure closure defects. In further studies to identify molecular therapies for this common congenital malformation, we now examine the effects of anti-apoptotic compounds in zebrafish models of ocular coloboma in vivo.
METHODS: Two ocular coloboma zebrafish lines (pax2.1/noi(tu29a) and lamb1/gup(m189)) were exposed to diferuloylmethane (curcumin) or benzyloxycarbonyl-Val-Ala-Asp(Ome)-fluoromethylketone (zVAD-fmk; a pan-caspase inhibitor) for up to 8 days post-fertilization. The effects of these compounds were assessed by morphology, histology, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and western blot analysis.
RESULTS: The size of the coloboma in gup zebrafish mutants treated with diferuloylmethane was greatly reduced. In treated mutants a reduction in TUNEL staining and a 67% decrease in activated caspase-3 protein were observed. The release of cytochrome c from the mitochondria into the cytosol was reduced fourfold by in vivo diferuloylmethane treatment, suggesting that the drug was acting to inhibit the intrinsic apoptotic pathway. Inhibition of caspases directly with zVAD-fmk also resulted in a similar reduction in coloboma phenotype. Treatment with either diferuloylmethane or zVAD-fmk resulted in a statistically significant 1.4 fold increase in length of survival of these mutant zebrafish (p<0.001), which normally succumb to the lethal genetic mutation. In contrast, the coloboma phenotype in noi zebrafish mutants did not respond to either diferuloylmethane or zVAD-fmk exposure, even though inhibition of apoptotic cell death was observed by a reduction in TUNEL staining.
CONCLUSIONS: The differential sensitivity to anti-apoptotic agents in lamb1-deficient and pax2.1-deficient zebrafish models, suggests that apoptotic cell death is not a final common pathway in all ocular coloboma genotypes. When considering anti-cell death therapies for ocular colobomatous defects attention should be paid to the genotype under investigation.
Cheryl Y Gregory-Evans; Mariya Moosajee; Xianghong Shan; Kevin Gregory-Evans
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2011-06-04
Journal Detail:
Title:  Molecular vision     Volume:  17     ISSN:  1090-0535     ISO Abbreviation:  Mol. Vis.     Publication Date:  2011  
Date Detail:
Created Date:  2011-06-16     Completed Date:  2011-10-26     Revised Date:  2013-06-28    
Medline Journal Info:
Nlm Unique ID:  9605351     Medline TA:  Mol Vis     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1473-84     Citation Subset:  IM    
Department of Ophthalmology and Visual Sciences, University of British Columbia, Vancouver BC, Canada.
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms
Amino Acid Chloromethyl Ketones / pharmacology
Blotting, Western
Caspase 3 / genetics,  metabolism
Cell Death / drug effects,  genetics*
Coloboma* / embryology,  genetics,  metabolism,  pathology
Curcumin / pharmacology*
Cytochromes c / analysis
Disease Models, Animal
Dose-Response Relationship, Drug
Embryo, Nonmammalian
Eye / embryology,  metabolism*,  pathology
Genetic Variation
In Situ Nick-End Labeling
Longevity / drug effects
Mitochondria / drug effects,  metabolism
Zebrafish / embryology,  genetics*,  metabolism
Zebrafish Proteins / genetics,  metabolism
Grant Support
VIH-105440//ASPE HHS; //Canadian Institutes of Health Research
Reg. No./Substance:
0/Amino Acid Chloromethyl Ketones; 0/Zebrafish Proteins; 0/benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; 458-37-7/Curcumin; 9007-43-6/Cytochromes c; EC 3.4.22.-/Caspase 3

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

Previous Document:  Age-related changes in the water-soluble lens protein composition of Wistar and accelerated-senescen...
Next Document:  A novel MERTK deletion is a common founder mutation in the Faroe Islands and is responsible for a hi...