Document Detail


Analysis of photoreceptor rod outer segment phagocytosis by RPE cells in situ.
MedLine Citation:
PMID:  23150373     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Counting rhodopsin-positive phagosomes residing in the retinal pigment epithelium (RPE) in the eye at different times of day allows a quantitative assessment of engulfment and digestion phases of diurnal RPE phagocytosis, which efficiently clears shed photoreceptor outer segment fragments (POS) from the neural retina. Comparing such activities among age- and background-matched experimental wild-type and mutant mice or rats serves to identify roles for specific proteins in the phagocytic process. Here, we describe experimental procedures for mouse eye harvest, embedding, sectioning, immunofluorescence labeling of rod POS phagosomes in RPE cells in sagittal eye sections, imaging of POS phagosomes in the RPE by laser scanning confocal microscopy, and POS quantification.
Authors:
Saumil Sethna; Silvia C Finnemann
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  935     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-15     Completed Date:  2013-04-17     Revised Date:  2014-01-10    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  245-54     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Fluorescent Antibody Technique / methods
Mice
Microscopy, Confocal / methods
Microtomy / methods
Phagocytosis*
Rats
Retinal Pigment Epithelium / cytology*,  metabolism,  ultrastructure
Rod Cell Outer Segment / metabolism,  ultrastructure*
Tissue Embedding / methods
Grant Support
ID/Acronym/Agency:
EY013295/EY/NEI NIH HHS; R01 EY013295/EY/NEI NIH HHS
Comments/Corrections

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