Expression and localisation of apical junctional complex proteins in lens epithelial cells

Yuki Sugiyama, Alan R. Prescott, Frederique M. D. Tholozan, Shigeo Ohno, Roy A. Quinlan

    Research output: Contribution to journalArticle

    14 Citations (Scopus)

    Abstract

    The lens epithelium possesses an apical junctional complex (AJC) comprising adherens and tight junctions (AJs and TJs) and yet several key structural components and associated regulatory proteins have not been identified or localised in these cells. Here we determine the subcellular distribution of the archetypal TJ markers (ZO-1, claudin-1, and occludin) and TJ-associated cell polarity proteins (aPKC, Par3 and Par6 beta) with AJ markers, E- and N-cadherin. As seen in other polarised epithelia, all these markers were located by confocal immunofluorescence microscopy to the apical ends of the lateral plasma membranes of bovine lens epithelial cells at sites of cell-cell interaction. Using immunoelectron microscopy, we show that ZO-1 concentrated at "kissing points" between neighbouring cells and these data, when taken in the context of our confocal immunofluorescence microscopy and blotting data, suggest the presence of TJs within the AJC. Likewise, immunogold labelling for E-cadherin identified AJs within these AJCs. We also report aPKC immunogold labelling localised to the AJC. These data show that the AJC of lens epithelial cells are a composite of TJs and AJs. (C) 2008 Elsevier Ltd. All rights reserved.

    Original languageEnglish
    Pages (from-to)64-70
    Number of pages7
    JournalExperimental Eye Research
    Volume87
    Issue number1
    DOIs
    Publication statusPublished - Jul 2008

    Keywords

    • lens
    • tight junction
    • adherens junction
    • cell polarity
    • aPKC
    • Par
    • electron microscopy
    • TIGHT JUNCTIONS
    • CHICKEN LENS
    • FIBER CELLS
    • POLARITY
    • PERMEABILITY
    • INVOLVEMENT
    • MICROSCOPY
    • PAR-3
    • ZO-1
    • FROG

    Cite this

    Sugiyama, Yuki ; Prescott, Alan R. ; Tholozan, Frederique M. D. ; Ohno, Shigeo ; Quinlan, Roy A. / Expression and localisation of apical junctional complex proteins in lens epithelial cells. In: Experimental Eye Research. 2008 ; Vol. 87, No. 1. pp. 64-70.
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    Expression and localisation of apical junctional complex proteins in lens epithelial cells. / Sugiyama, Yuki; Prescott, Alan R.; Tholozan, Frederique M. D.; Ohno, Shigeo; Quinlan, Roy A.

    In: Experimental Eye Research, Vol. 87, No. 1, 07.2008, p. 64-70.

    Research output: Contribution to journalArticle

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    AU - Sugiyama, Yuki

    AU - Prescott, Alan R.

    AU - Tholozan, Frederique M. D.

    AU - Ohno, Shigeo

    AU - Quinlan, Roy A.

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    N2 - The lens epithelium possesses an apical junctional complex (AJC) comprising adherens and tight junctions (AJs and TJs) and yet several key structural components and associated regulatory proteins have not been identified or localised in these cells. Here we determine the subcellular distribution of the archetypal TJ markers (ZO-1, claudin-1, and occludin) and TJ-associated cell polarity proteins (aPKC, Par3 and Par6 beta) with AJ markers, E- and N-cadherin. As seen in other polarised epithelia, all these markers were located by confocal immunofluorescence microscopy to the apical ends of the lateral plasma membranes of bovine lens epithelial cells at sites of cell-cell interaction. Using immunoelectron microscopy, we show that ZO-1 concentrated at "kissing points" between neighbouring cells and these data, when taken in the context of our confocal immunofluorescence microscopy and blotting data, suggest the presence of TJs within the AJC. Likewise, immunogold labelling for E-cadherin identified AJs within these AJCs. We also report aPKC immunogold labelling localised to the AJC. These data show that the AJC of lens epithelial cells are a composite of TJs and AJs. (C) 2008 Elsevier Ltd. All rights reserved.

    AB - The lens epithelium possesses an apical junctional complex (AJC) comprising adherens and tight junctions (AJs and TJs) and yet several key structural components and associated regulatory proteins have not been identified or localised in these cells. Here we determine the subcellular distribution of the archetypal TJ markers (ZO-1, claudin-1, and occludin) and TJ-associated cell polarity proteins (aPKC, Par3 and Par6 beta) with AJ markers, E- and N-cadherin. As seen in other polarised epithelia, all these markers were located by confocal immunofluorescence microscopy to the apical ends of the lateral plasma membranes of bovine lens epithelial cells at sites of cell-cell interaction. Using immunoelectron microscopy, we show that ZO-1 concentrated at "kissing points" between neighbouring cells and these data, when taken in the context of our confocal immunofluorescence microscopy and blotting data, suggest the presence of TJs within the AJC. Likewise, immunogold labelling for E-cadherin identified AJs within these AJCs. We also report aPKC immunogold labelling localised to the AJC. These data show that the AJC of lens epithelial cells are a composite of TJs and AJs. (C) 2008 Elsevier Ltd. All rights reserved.

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