title: FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes.
abstract: Proper development and tissue maintenance requires cell-cell adhesion structures, which serve diverse and crucial roles in tissue morphogenesis. Epithelial tissues have three main types of cell-cell junctions: tight junctions, which play a major role in barrier formation, and adherens junctions and desmosomes, which provide mechanical stability and organize the underlying cytoskeleton. Our current understanding of adhesion function is hindered by a lack of tools and methods to image junctions in mammals. To better understand the dynamics of adhesion in tissues we have created a knock-in ZO-1-GFP mouse and a BAC-transgenic mouse expressing desmoplakin I-GFP. We performed fluorescence recovery after photobleaching (FRAP) experiments to quantify the turnover rates of the tight junction protein ZO-1, the adherens junction protein E-cadherin, and the desmosomal protein desmoplakin in the epidermis. Proteins at each type of junction are remarkably stable in the epidermis, in contrast to the high observed mobility of E-cadherin and ZO-1 at adherens junctions and tight junctions, respectively, in cultured cells. Our data demonstrate that there are additional mechanisms for stabilizing junctions in tissues that are not modeled by cell culture.
authors: Department of Cell Biology, Duke University Medical Center, Durham, North Carolina, United State of America.
authors: Henry P
authors: HP
authors: Foote
authors: Kaelyn D
authors: KD
authors: Sumigray
authors: Terry
authors: T
authors: Lechler
mesh_terms: D022005:Adherens Junctions / Q000378:metabolism; D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion; D002478:Cells, Cultured; D051180:Desmoplakins / Q000378:metabolism; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism*; D036681:Fluorescence Recovery After Photobleaching; D055879:Gene Knock-In Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology* / Q000378:metabolism*; D051379:Mice; D020033:Protein Isoforms / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D015820:Cadherins; D051180:Desmoplakins; D020033:Protein Isoforms; D062826:Zonula Occludens-1 Protein; D049452:Green Fluorescent Proteins
references: J Cell Biol. 2013 Apr 1;201(1):131-43
references: 23547031
references: J Cell Biol. 2007 Jan 15;176(2):147-54
references: 17227889
references: Development. 1988 Apr;102(4):639-55
references: 3048970
references: Nat Cell Biol. 2001 Dec;3(12):1076-85
references: 11781569
references: Proc Natl Acad Sci U S A. 2010 May 4;107(18):8237-41
references: 20404178
references: Nat Cell Biol. 2012 Sep;14(9):935-43
references: 22885771
references: J Cell Biol. 2002 Mar 18;156(6):1099-111
references: 11889141
references: Cell. 1996 Feb 9;84(3):345-57
references: 8608588
references: J Cell Biol. 2012 Oct 29;199(3):513-25
references: 23091070
references: Nat Cell Biol. 2010 Jun;12(6):533-42
references: 20453849
references: Nature. 2008 Jun 5;453(7196):751-6
references: 18480755
references: J Cell Biol. 2008 May 19;181(4):683-95
references: 18474622
references: J Invest Dermatol. 1987 Sep;89(3):253-63
references: 2442269
references: Biochim Biophys Acta. 2013 Mar;1830(3):2690-700
references: 23671930
references: J Invest Dermatol. 2007 Nov;127(11):2499-515
references: 17934502
references: Proc Natl Acad Sci U S A. 2008 Oct 7;105(40):15405-10
references: 18809908
references: Cell. 2010 Oct 1;143(1):134-44
references: 20887898
references: Nat Rev Mol Cell Biol. 2005 Aug;6(8):622-34
references: 16025097
references: J Invest Dermatol. 2011 Mar;131(3):706-18
references: 21160493
references: Curr Opin Cell Biol. 2002 Oct;14(5):531-6
references: 12231346
references: J Cell Biol. 2011 Aug 22;194(4):631-42
references: 21844209
references: J Biol Chem. 2009 Jul 31;284(31):21036-46
references: 19478092
references: J Cell Sci. 2011 Dec 1;124(Pt 23):4001-13
references: 22159415
references: Mol Biol Cell. 2008 Jun;19(6):2465-75
references: 18353970
references: Am J Physiol Cell Physiol. 2004 Jun;286(6):C1213-28
references: 15151915