| Molecular and biochemical characterization of folate transport proteins in retinal Müller cells. | |
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MedLine Citation:
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PMID: 20053979 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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PURPOSE: To analyze the mechanisms of folate uptake in retinal Müller cells. METHODS: RT-PCR and Western blot analysis were performed in freshly isolated neural retina and RPE/eyecup, primary mouse Müller cells, and rMC-1 cells for the three known folate transport proteins folate receptor alpha (FRalpha), proton-coupled folate transporter (PCFT), and reduced folate carrier (RFC). Laser scanning confocal microscopy (LSCM) and immunoelectron microscopy were used to determine the subcellular location of FRalpha and PCFT in primary Müller cells. The pH dependence of the uptake of [(3)H]-methyltetrahydrofolate ([(3)H]-MTF) was assayed in Müller cells in the presence/absence of thiamine pyrophosphate, an inhibitor of RFC. RESULTS: FRalpha and PCFT are expressed abundantly in the retina in several cell layers, including the inner nuclear layer; they are present in primary mouse Müller cells and rMC-1 cells. LSCM localized these proteins to the plasma membrane, nuclear membrane, and perinuclear region. Immunoelectron microscopic studies revealed the colocalization of FRalpha and PCFT on the plasma membrane and nuclear membrane and within endosomal structures. Müller cell uptake of [(3)H]-MTF was robust at pH 5.0 to 6.0, consistent with PCFT activity, but also at neutral pH, reflecting RFC function. RFC was expressed in mouse Müller cells that had been allowed to proliferate in culture, but not in freshly isolated primary cells. CONCLUSIONS: FRalpha and PCFT are expressed in retinal Müller cells and colocalize in the endosomal compartment, suggesting that the two proteins may work coordinately to mediate folate uptake. The unexpected finding of RFC expression and activity in cultured Müller cells may reflect the upregulation of this protein under proliferative conditions. |
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Authors:
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B Renee Bozard; Preethi S Ganapathy; Jennifer Duplantier; Barbara Mysona; Yonju Ha; Penny Roon; Robert Smith; I David Goldman; Puttur Prasad; Pamela M Martin; Vadivel Ganapathy; Sylvia B Smith |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural Date: 2010-01-06 |
Journal Detail:
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Title: Investigative ophthalmology & visual science Volume: 51 ISSN: 1552-5783 ISO Abbreviation: Invest. Ophthalmol. Vis. Sci. Publication Date: 2010 Jun |
Date Detail:
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Created Date: 2010-05-20 Completed Date: 2010-06-11 Revised Date: 2011-05-11 |
Medline Journal Info:
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Nlm Unique ID: 7703701 Medline TA: Invest Ophthalmol Vis Sci Country: United States |
Other Details:
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Languages: eng Pagination: 3226-35 Citation Subset: IM |
Affiliation:
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Department of Cellular Biology and Anatomy, Medical College of Georgia, Augusta, Georgia 30912-2000, USA. |
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Blotting, Western Cells, Cultured Endosomes / metabolism Folate Receptor 1 Gene Expression Regulation / physiology Hydrogen-Ion Concentration Membrane Transport Proteins / genetics, metabolism*, ultrastructure Mice Mice, Inbred BALB C Microscopy, Confocal Microscopy, Immunoelectron Neuroglia / drug effects, metabolism*, ultrastructure Proton-Coupled Folate Transporter Receptors, Cell Surface / genetics, metabolism*, ultrastructure Reduced Folate Carrier Protein Retina / metabolism Reverse Transcriptase Polymerase Chain Reaction Tetrahydrofolates / metabolism* Thiamine Pyrophosphate / pharmacology |
| Grant Support | |
ID/Acronym/Agency:
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R01 EY 012830/EY/NEI NIH HHS; R01 EY012830-09/EY/NEI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Folate Receptor 1; 0/Folr1 protein, mouse; 0/Membrane Transport Proteins; 0/Proton-Coupled Folate Transporter; 0/Receptors, Cell Surface; 0/Reduced Folate Carrier Protein; 0/Slc19a1 protein, mouse; 0/Slc46a1 protein, mouse; 0/Tetrahydrofolates; 134-35-0/5-methyltetrahydrofolate; 154-87-0/Thiamine Pyrophosphate |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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