Document Detail


Recoverin in cultured human retinoblastoma cells: enhanced expression during morphological differentiation.
MedLine Citation:
PMID:  8666979     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Recoverin is a calcium-binding protein expressed in retinal photoreceptors. It appears to delay the termination of the phototransduction cascade by blocking the phosphorylation of photoexcited rhodopsin. The goal of this study was to determine if recoverin mRNA and protein are expressed in cultured human Y79 retinoblastoma cells, so that this cell line could be used as a model to study the mechanism of recoverin gene expression in the retina. A cDNA encoding human recoverin was PCR cloned and used for prokaryotic expression of recoverin protein. Polyclonal antibodies raised against pure recombinant recoverin were used for western blotting and immunocytochemistry of Y79 cells grown as attachment cultures in the presence of the differentiating agents dibutyryl cyclic AMP (dbcAMP) or butyrate. Northern blot analysis was performed on mRNA extracted from Y79 cells that were also treated with the differentiating agents. In Y79 cell monolayer cultures, recoverin was immunolocalized to the cell cytoplasm, and immunoreactivity was increased dramatically by the addition of 2 mM butyrate to the culture medium. Butyrate treatment also caused an increase in the development of neurite-like cellular processes. Addition of 4 mM dbcAMP resulted in a moderate increase in both recoverin immunoreactivity and number of cellular processes. Western and northern blots of butyrate and dbcAMP-treated Y79 cell cultures demonstrated an increase in recoverin protein and RNA expression, respectively, comparable with that observed with immunocytochemistry. These data suggest that, under the influence of the differentiating agent butyrate, Y79 cells exhibit an increase in expression of the photoreceptor protein recoverin and a concomitant morphological differentiation toward a neuronal phenotype.
Authors:
A F Wiechmann
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  67     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  1996 Jul 
Date Detail:
Created Date:  1996-08-06     Completed Date:  1996-08-06     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  105-10     Citation Subset:  IM    
Affiliation:
Department of Anatomy and Neurobiology, Boston University School of Medicine, Boston, 02118, USA.
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MeSH Terms
Descriptor/Qualifier:
Antibody Specificity
Antigens, Neoplasm / analysis,  genetics,  immunology
Blotting, Northern
Blotting, Western
Bucladesine / pharmacology
Butyric Acid
Butyric Acids / pharmacology
Calcium-Binding Proteins / analysis,  genetics*,  immunology
Cell Differentiation / physiology
Eye Proteins*
Gene Expression / drug effects
Hippocalcin
Humans
Immunohistochemistry
Lipoproteins*
Nerve Tissue Proteins*
RNA, Messenger / metabolism
Recoverin
Retinoblastoma
Tumor Cells, Cultured / chemistry,  cytology,  physiology
Tumor Markers, Biological / analysis*
Grant Support
ID/Acronym/Agency:
EY81001/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, Neoplasm; 0/Butyric Acids; 0/Calcium-Binding Proteins; 0/Eye Proteins; 0/Lipoproteins; 0/Nerve Tissue Proteins; 0/RCVRN protein, human; 0/RNA, Messenger; 0/Tumor Markers, Biological; 107-92-6/Butyric Acid; 135844-11-0/Recoverin; 149223-81-4/Hippocalcin; 362-74-3/Bucladesine

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


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