FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes.
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...
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doaj-54bbb25d818a426087c789a368abb3882020-11-25T01:00:27ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0188e7149110.1371/journal.pone.0071491FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes.Henry P FooteKaelyn D SumigrayTerry LechlerProper 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.http://europepmc.org/articles/PMC3747223?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Henry P Foote Kaelyn D Sumigray Terry Lechler |
spellingShingle |
Henry P Foote Kaelyn D Sumigray Terry Lechler FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. PLoS ONE |
author_facet |
Henry P Foote Kaelyn D Sumigray Terry Lechler |
author_sort |
Henry P Foote |
title |
FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. |
title_short |
FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. |
title_full |
FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. |
title_fullStr |
FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. |
title_full_unstemmed |
FRAP analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. |
title_sort |
frap analysis reveals stabilization of adhesion structures in the epidermis compared to cultured keratinocytes. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2013-01-01 |
description |
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. |
url |
http://europepmc.org/articles/PMC3747223?pdf=render |
work_keys_str_mv |
AT henrypfoote frapanalysisrevealsstabilizationofadhesionstructuresintheepidermiscomparedtoculturedkeratinocytes AT kaelyndsumigray frapanalysisrevealsstabilizationofadhesionstructuresintheepidermiscomparedtoculturedkeratinocytes AT terrylechler frapanalysisrevealsstabilizationofadhesionstructuresintheepidermiscomparedtoculturedkeratinocytes |
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