Interactions of Recombinant Mouse Erythrocyte Transglutaminase with Membrane Skeletal Proteins |
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Authors: | Edgar Gutierrez L Amy Sung |
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Institution: | (1) Department of Anatomy and Cell Biology, University of Western Ontario, London, N6A-5C1, Ontario, Canada |
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Abstract: | In this study, we chose a differentiation-competent rat epidermal keratinocyte (REK) cell line to examine the role of Cx26
and disease-linked Cx26 mutants in organotypic epidermal differentiation. First, we generated stable REK cell lines expressing
three skin disease-linked mutants (G59A, D66H and R75W). Second, we used an RNAi approach to knock down the expression of
Cx26 in REKs. Interestingly, the three-dimensional (3D) architecture of the organotypic epidermis altered the intracellular
spatial distribution of the mutants in comparison to 2D cultured REKs, highlighting the importance of using organotypic cultures.
Unexpectedly, the presence of disease-linked mutants or the overexpression of wild-type Cx26 had little effect on the differentiation
of the organotypic epidermis as determined by the architecture of the epidermis, expression of molecular markers indicative
of epidermis differentiation (keratin 10, keratin 14, involucrin, loricrin) and stratification/cornification of the epidermis.
Likewise, organotypic epidermis continued to differentiate normally upon Cx26 knockdown. While Cx26 has been reported to be
upregulated during wound healing, no reduction in wound closure was observed in 2D REK cultures that expressed loss-of-function,
dominant Cx26 mutants. In conclusion, we demonstrate that gain or loss of Cx26 function does not disrupt organotypic epidermal
differentiation and offer insights into why patients harboring Cx26 mutations do not frequently present with more severe disease
that encompasses thin skin.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Cx26 Epidermis Mutant Differentiation Disease |
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