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AMPK Activation Promotes Tight Junction Assembly in Intestinal Epithelial Caco-2 Cells

Abstract : The AMP-activated protein kinase (AMPK) is principally known as a major regulator of cellular energy status, but it has been recently shown to play a key structural role in cell-cell junctions. The aim of this study was to evaluate the impact of AMPK activation on the reassembly of tight junctions in intestinal epithelial Caco-2 cells. We generated Caco-2 cells invalidated for AMPK α1/α2 (AMPK dKO) by CRISPR/Cas9 technology and evaluated the effect of the direct AMPK activator 991 on the reassembly of tight junctions following a calcium switch assay. We analyzed the integrity of the epithelial barrier by measuring the trans-epithelial electrical resistance (TEER), the paracellular permeability, and quantification of zonula occludens 1 (ZO-1) deposit at plasma membrane by immunofluorescence. Here, we demonstrated that AMPK deletion induced a delay in tight junction reassembly and relocalization at the plasma membrane during calcium switch, leading to impairments in the establishment of TEER and paracellular permeability. We also showed that 991-induced AMPK activation accelerated the reassembly and reorganization of tight junctions, improved the development of TEER and paracellular permeability after calcium switch. Thus, our results show that AMPK activation ensures a better recovery of epithelial barrier function following injury.
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Submitted on : Wednesday, October 23, 2019 - 7:12:11 PM
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Séverine Olivier, Jocelyne Leclerc, Adrien Grenier, Marc Foretz, Jérôme Tamburini, et al.. AMPK Activation Promotes Tight Junction Assembly in Intestinal Epithelial Caco-2 Cells. International Journal of Molecular Sciences, MDPI, 2019, 20 (20), pp.5171. ⟨10.3390/ijms20205171⟩. ⟨inserm-02330186⟩



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