Comment[ArrayExpressAccession]	E-GEOD-48443
MAGE-TAB Version	1.1																	
Public Release Date	2014-04-06
Investigation Title	Knockout mice reveal key roles for claudin-18 in alveolar fluid homeostasis.																	
Comment[Submitted Name]	Knockout mice reveal key roles for claudin-18 in alveolar fluid homeostasis.
Experiment Description	Claudin proteins are major constituents of epithelial and endothelial tight junctions (TJ), where they serve as regulators of paracellular permeability to ions and solutes. Claudin-18, a member of the large claudin family, is highly expressed in lung epithelium. To elucidate the role of claudin-18 in alveolar epithelial barrier function and fluid homeostasis, we generated claudin-18 knockout (C18 KO) mice. Increased alveolar fluid clearance (AFC) observed in C18 KO mice may have accounted for absence of lung edema despite increased alveolar solute permeability compared to wild type (WT) controls. Higher AFC in C18 KO mice was associated with higher Na-K-ATPase activity and increased expression of the Na-K-ATPase β1 subunit compared to WT controls. Consistent with in vivo findings, alveolar epithelial cell (AEC) monolayers derived from C18 KO mice exhibited lower transepithelial electrical resistance (RT) accompanied by increased solute and ion permeability without changes in ion selectivity. Expression of claudin-3 and claudin-4 was markedly increased in whole lung and in freshly isolated AEC from C18 KO mice, while claudin-5 was unchanged. In contrast, occludin, another major component of the TJ complex, was significantly decreased in C18 KO lung. Further analysis revealed rearrangements in the F-actin cytoskeleton in C18 KO MAECM. These findings demonstrate a crucial non-redundant role for claudin-18 in regulation of alveolar epithelial tight junction composition and permeability to ions and solutes.  Importantly, increased AFC in C18 KO mice identifies additional roles for claudin-18 in alveolar fluid homeostasis beyond its direct contributions to barrier properties of the alveolar epithelium. Animals with a ubiquitous knockout (C18 KO) were obtained by crossing mice harboring a conditional (floxed) allele of claudin-18 (Cldn18F/F) with CMV-cre deleter mice to delete exons 2 and 3 by Cre/loxP recombination.																	
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl																
Person Last Name	Marconett	Li	Flodby	Luo	Kage	Sipos	Gao	Beard	Marconett	DeMaio	Kim	Kim	Laird-Offringa	Minoo	Liebler	Zhou	Crandall	Borok
Person First Name	Crystal	Guanglei	Per	Jiao	Hidenori	Arnold	Danping	LaMonta	Crystal	Lucas	Yong	Kwang-Jin	Ite	Parviz	Janice	Beiyun	Edward	Zea
Person Mid Initials	N							L	N		H		A		M		D	
Person Email	cmarcone@usc.edu																	
Person Affiliation	University of Southern California																	
Person Phone	323-865-0654																	
Person Address	Biochemistry & Molecular Biology, University of Southern California, 1441 Eastlake Ave NOR 6420, Los Angeles, CA, USA																	
Person Roles	submitter																	
Protocol Name	P-GSE48443-1	P-GSE48443-5	P-GSE48443-6	P-GSE48443-2	P-GSE48443-3	P-GSE48443-4	P-GSE48443-7											
Protocol Description	Background correction using variance stabilizing transformation (VST) and quantile normalization were performed using the LUMI package (v.1.14.0) in R v.2.11.1 (2010-05-31). ID_REF =  VALUE = Normalized signal intensity Detection Pval = 	IlluminaTotalPrep RNA kit	Standard Illumina hybridization protocol	No treatment	Whole lungs were homogenized in Trizol with a Polytron.	1μg of RNA from four replicates each from wildtype and claudin-18 knockout whole lung was converted into cRNA and was used for Illumina mouseRef-8 v2 whole genome expression analysis. Expression arrays were carried out at the Southern California Genotyping Consortium (SCGC) at the University of California, Los Angeles.	BeadStudio was used to convert images to raw signal data.											
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	sample treatment protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol											
Experimental Factor Name	GENOTYPE																	
Experimental Factor Type	genotype																	
Publication Title	Knockout mice reveal key roles for claudin 18 in alveolar barrier properties and fluid homeostasis.																	
Publication Author List	Li G, Flodby P, Luo J, Kage H, Sipos A, Gao D, Ji Y, Beard LL, Marconett CN, Demaio L, Kim YH, Kim KJ, Laird-Offringa IA, Minoo P, Liebler JM, Zhou B, Crandall ED, Borok Z																	
PubMed ID	24588076																	
Publication DOI	10.1165/rcmb.2013-0353OC																	
Comment[SecondaryAccession]	GSE48443																	
Comment[GEOReleaseDate]	2014-04-06																	
Comment[ArrayExpressSubmissionDate]	2013-07-01																	
Comment[GEOLastUpdateDate]	2014-04-07																	
Comment[AEExperimentType]	transcription profiling by array																	
Comment[AdditionalFile:Data1]	GSE48443_non_normalized.txt																	
SDRF File	E-GEOD-48443.sdrf.txt