Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
5
pubmed:dateCreated
2006-9-25
pubmed:abstractText
Corneal endothelium secretes HCO(3)(-) from basolateral (stroma) to apical (anterior chamber) compartments. Apical HCO(3)(-) permeability can be enhanced by increasing [Ca(2+)](i). We hypothesized that the bovine calcium-activated chloride channel 1 (bCLCA1), shown previously by PCR screening to be expressed in corneal endothelium, is involved in Ca(2+) activated apical HCO(3)(-) permeability. bCLCA1 expression in cultured bovine corneal endothelial cells (CBCEC) was examined by in situ hybridization analysis, immunoblotting, immunofluorescence and confocal microscopy. Rabbit polyclonal antibodies were generated using a 14 aa polypeptide (417-430) from the predicted sequence of bCLCA1. The small interference RNA (siRNA) knock down technique was used to evaluate the functional involvement of bCLCA1 in apical HCO(3)(-) permeability. In situ hybridization confirmed prominent bCLCA1-specific mRNA expression in CBCEC. bCLCA1 antiserum detected the heterologously expressed bCLCA1 in HEK293 cells and a 90kDa band in CBCEC, which was absent when using the pre-immune serum or antigen absorption of serum. Immunofluoresence staining with anti-bCLCA1 antibody and confocal microscopy indicates an apical membrane location in CBCEC. In CBCEC transfected with bCLCA1 specific siRNA, bCLCA1 expression was reduced by 80%, while transfection with siControl scrambled sequence had no effect. Increasing [Ca(i)(2+)] by application of ATPgammaS or cyclopiazonic acid (CPA) increased apical HCO(3)(-) permeability in siControl transfected CBCEC, while having no effect on apical HCO(3)(-) permeability in bCLCA1 specific siRNA transfected cells. Baseline HCO(3)(-) permeability, however, was not different between controls and siRNA treated cells. We conclude that the calcium-activated chloride channel (bCLCA1) is expressed in bovine corneal endothelial cells and can contribute to Ca(2+) dependent apical HCO(3)(-) permeability, but not resting permeability, across the corneal endothelium.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-10070252, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-10484341, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11029313, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11071307, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11071308, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11402373, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11880256, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11917096, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-11923259, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-12504118, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-12570711, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-12597924, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-12867361, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-12907161, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-1371273, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-1383206, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-1399410, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-14895848, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-14960417, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15381038, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15548570, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15635293, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15709976, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15862176, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15919655, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15978249, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-15987802, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-1705933, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-4215880, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-5016991, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-7084312, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-7822137, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-7929397, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-828203, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-8537359, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-9678416, http://linkedlifedata.com/resource/pubmed/commentcorrection/16899243-9856782
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0014-4835
pubmed:author
pubmed:issnType
Print
pubmed:volume
83
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1215-24
pubmed:dateRevised
2011-7-28
pubmed:meshHeading
pubmed:year
2006
pubmed:articleTitle
Molecular expression and functional involvement of the bovine calcium-activated chloride channel 1 (bCLCA1) in apical HCO3- permeability of bovine corneal endothelium.
pubmed:affiliation
School of Optometry, Indiana University, 800 East Atwater Avenue, Bloomington, IN 47405, USA.
pubmed:publicationType
Journal Article, Research Support, N.I.H., Extramural