Document Detail


Thy1 associates with the cation channel subunit HCN4 in adult rat retina.
MedLine Citation:
PMID:  22281825     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: The membrane expression and gene promoter of the glycosylphosphatidylinositol (GPI)-anchored protein Thy1 have been widely used to examine the morphology and distribution of retinal ganglion cells in normal eyes and disease models. However, it is not known how adult mammalian retinal neurons use Thy1. Because Thy1 is not a membrane-spanning protein and, instead, complexes with structural and signaling proteins in other tissues, the aim of this study was to find protein partners of retinal Thy1.
METHODS: Coimmunoprecipitation, immunohistochemistry, confocal imaging, and patch-clamp recording were used to test for association of Thy1 and HCN4, a cation channel subunit, in adult rat retina.
RESULTS: Hyperpolarization of cells immunopanned by an anti-Thy1 antibody activated HCN channels. Confocal imaging showed that individual somata in the ganglion cell layer bound antibodies against Thy1 and HCN4, that the majority of these bindings colocalized, and that some of the immunopositive cells also bound antibody against a ganglion cell marker (Brn3a). Consistent with these results, Thy1 and HCN4 were coimmunoprecipitated by magnetic beads coated with either anti-Thy1 antibody or anti-HCN4 antibody. In control experiments, beads coated with these antibodies did not immunoprecipitate a photoreceptor rim protein (ABCR) and uncoated beads did not immunoprecipitate either Thy1 or HCN4.
CONCLUSIONS: This is the first report that Thy1 colocalizes and coimmunoprecipitates with a membrane-spanning protein in retina, that Thy1 complexes with an ion channel protein in any tissue, and that a GPI-anchored protein associates with an HCN channel subunit protein.
Authors:
Gloria J Partida; Tyler W Stradleigh; Genki Ogata; Iv Godzdanker; Andrew T Ishida
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Publication Detail:
Type:  Journal Article; Research Support, American Recovery and Reinvestment Act; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-03-26
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  53     ISSN:  1552-5783     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2012 Mar 
Date Detail:
Created Date:  2012-03-27     Completed Date:  2012-05-09     Revised Date:  2013-06-26    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1696-703     Citation Subset:  IM    
Affiliation:
Section of Neurobiology, Physiology, and Behavior, University ofCalifornia, Davis, California 95616-8519, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antigens, Thy-1 / metabolism*
Female
Immunoprecipitation
Patch-Clamp Techniques
Potassium Channels / metabolism*
Rats
Rats, Long-Evans
Retinal Ganglion Cells / cytology,  metabolism*
Grant Support
ID/Acronym/Agency:
EY08120/EY/NEI NIH HHS; EY08120-17S1/EY/NEI NIH HHS; EY08120-20S1/EY/NEI NIH HHS; P30 EY12576/EY/NEI NIH HHS; R25 56765//PHS HHS; T32 EY015387/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/Antigens, Thy-1; 0/HCN4 protein, rat; 0/Potassium Channels
Comments/Corrections

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