Document Detail


Expression of aquaporin-1 in human trabecular meshwork cells: role in resting cell volume.
MedLine Citation:
PMID:  11431445     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: Drainage of aqueous humor from the human eye appears dependent on intracellular volume of trabecular meshwork (TM) cells, the predominant cell type of the human outflow pathway. Thus, the modulation of water and solute flux across the plasma membrane of TM cells is predicted to be an important factor in regulating outflow facility. Aquaporin (AQP)-1 is a hexahelical integral membrane protein that functions as a regulated channel for water and cations in fluid-secreting and -absorbing tissues. AQP1 is present in many tissues of the human eye, including the TM; however, its role in outflow facility is unknown. The purpose of the present study was twofold: to evaluate the prospect of manipulating AQP1 protein levels in TM cells using sense and antisense mRNA and to investigate the functional role of AQP1 in TM cells. METHODS: An adenovirus (AV) expression system was used to alter AQP1 protein levels. AQP1 protein expression was monitored using immunoblot analysis, and resting cell volume was measured by forward light scatter, electronic cell sizing, and [(14)C]-sucrose/urea equilibration. Permeability of TM monolayers to [(14)C]-sucrose was also assessed as an indirect evaluation of cell volume. RESULTS: AV-mediated gene transfer of AQP1 cDNA to TM cells resulted in a titer-dependent increase in recombinant AQP1, whereas transfer of antisense cDNA decreased native AQP1 protein by 71.7% +/- 5.5% (P < 0.01) after 5 days. A novel finding of this study is that mean resting volumes of AQP1(s) AV-infected TM cells in suspension were 8.7% +/- 3.0% greater (P < 0.05) than control cells. Conversely, AQP1 antisense (as) AV-infected cells had resting volumes 7.8% +/- 2.9% less than control cells (P < 0.05). Similar effects of AQP1 expression on resting cell volume were observed in TM monolayers. Consistent with this finding, paracellular permeability of AQP1(s) AV-infected TM monolayers to [(14)C]-sucrose decreased by 8.0% +/- 1.4% (P < 0.001). CONCLUSIONS: In addition to influencing the osmotic permeability of TM plasma membranes, the level of AQP1 protein expression influences resting intracellular volume and thus paracellular permeability of TM cell monolayers in vitro. These data suggest that AQP1 expression may affect outflow facility in vivo.
Authors:
W D Stamer; K Peppel; M E O'Donnell; B C Roberts; F Wu; D L Epstein
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Investigative ophthalmology & visual science     Volume:  42     ISSN:  0146-0404     ISO Abbreviation:  Invest. Ophthalmol. Vis. Sci.     Publication Date:  2001 Jul 
Date Detail:
Created Date:  2001-06-29     Completed Date:  2001-08-02     Revised Date:  2010-03-24    
Medline Journal Info:
Nlm Unique ID:  7703701     Medline TA:  Invest Ophthalmol Vis Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1803-11     Citation Subset:  IM    
Affiliation:
Department of Ophthalmology, The University of Arizona, 655 North Alvernon Way, Tucson, AZ 85711-1824, USA. dstamer@eyes.arizona.edu
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MeSH Terms
Descriptor/Qualifier:
Adenoviruses, Human / genetics
Adolescent
Aged
Aquaporin 1
Aquaporins / physiology*
Blood Group Antigens
Cell Membrane Permeability
Cell Size / physiology
Gene Expression
Gene Transfer Techniques
Humans
Immunoblotting
Infant, Newborn
Middle Aged
RNA / isolation & purification
RNA, Messenger / biosynthesis
Recombinant Proteins
Reverse Transcriptase Polymerase Chain Reaction
Trabecular Meshwork / cytology*,  metabolism
Grant Support
ID/Acronym/Agency:
F32 EY0685/EY/NEI NIH HHS; P30 EY05722/EY/NEI NIH HHS; R01 EY01894/EY/NEI NIH HHS
Chemical
Reg. No./Substance:
0/AQP1 protein, human; 0/Aquaporins; 0/Blood Group Antigens; 0/RNA, Messenger; 0/Recombinant Proteins; 146410-94-8/Aquaporin 1; 63231-63-0/RNA

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


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