Function of MYO7A in the Human RPE and the Validity of Shaker1 Mice as a Model for Usher Syndrome 1B

To investigate the function of MYO7A in human RPE cells and to test the validity of using shaker1 RPE in preclinical studies on therapies for Usher syndrome 1B by comparing human and mouse cells. MYO7A was localized by immunofluorescence. Primary cultures of human and mouse RPE cells were used to me...

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Bibliographic Details
Published in:Investigative ophthalmology & visual science 2010-02, Vol.51 (2), p.1130-1135
Main Authors: Gibbs, Daniel, Diemer, Tanja, Khanobdee, Kornnika, Hu, Jane, Bok, Dean, Williams, David S
Format: Article
Language:English
Subjects:
Aged, 80 and over
Animals
Blotting, Western
Cells, Cultured
Disease Models, Animal
Fluorescent Antibody Technique, Indirect
Gene Silencing
Humans
Male
Melanosomes - physiology
Mice
Mice, Inbred C57BL
Mice, Mutant Strains
Microscopy, Confocal
Myosins - physiology
Phagocytosis - physiology
Retinal Pigment Epithelium - metabolism
RNA Interference
Rod Cell Outer Segment - physiology
Transfection
Usher Syndromes - metabolism
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Summary:To investigate the function of MYO7A in human RPE cells and to test the validity of using shaker1 RPE in preclinical studies on therapies for Usher syndrome 1B by comparing human and mouse cells. MYO7A was localized by immunofluorescence. Primary cultures of human and mouse RPE cells were used to measure melanosome motility and rod outer segment (ROS) phagocytosis and digestion. MYO7A was knocked down in the human RPE cells by RNAi to test for a mutant phenotype in melanosome motility. The distribution of MYO7A in the RPE of human and mouse was found to be comparable, both in vivo and in primary cultures. Primary cultures of human RPE cells phagocytosed and digested ROSs with kinetics comparable to that of primary cultures of mouse RPE cells. Melanosome motility was also comparable, and, after RNAi knockdown, consisted of longer-range fast movements characteristic of melanosomes in shaker1 RPE. The localization and function of MYO7A in human RPE cells is comparable to that in mouse RPE cells. Although shaker1 retinas do not undergo degeneration, correction of mutant phenotypes in the shaker1 RPE represents a valid preclinical test for potential therapeutic treatments.
ISSN:0146-0404
1552-5783
1552-5783
DOI:10.1167/iovs.09-4032