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2016 ; 291
(47
): 24787-24799
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Hypotonic Stress-induced Down-regulation of Claudin-1 and -2 Mediated by
Dephosphorylation and Clathrin-dependent Endocytosis in Renal Tubular Epithelial
Cells
#MMPMID27733684
Fujii N
; Matsuo Y
; Matsunaga T
; Endo S
; Sakai H
; Yamaguchi M
; Yamazaki Y
; Sugatani J
; Ikari A
J Biol Chem
2016[Nov]; 291
(47
): 24787-24799
PMID27733684
show ga
Hypotonic stress decreased claudin-1 and -2 expression levels in renal tubular
epithelial HK-2 and Madin-Darby canine kidney cells. Here, we examined the
regulatory mechanism involved in this decrease. The hypotonicity-induced decrease
in claudin expression was inhibited by the following: SB202190, a p38 MAPK
inhibitor, but not by U0126, a MEK inhibitor; Go6983, a protein kinase C
inhibitor; or SP600125, a Jun N-terminal protein kinase inhibitor. Hypotonic
stress increased transepithelial electrical resistance, which was inhibited by
SB202190. The mRNA expression level of claudin-1 was decreased by hypotonic
stress but that of claudin-2 was not. Hypotonic stress decreased the protein
stability of claudin-1 and -2. The hypotonicity-induced decrease in claudin
expression was inhibited by the following: chloroquine, a lysosome inhibitor;
dynasore and monodansylcadaverine, clathrin-dependent endocytosis inhibitors; and
siRNA against clathrin heavy chain. Claudin-1 and -2 were mainly distributed in
the cytosol and tight junctions (TJs) in the chloroquine- and
monodansylcadaverine-treated cells, respectively. Hypotonic stress decreased the
phosphorylation levels of claudin-1 and -2, which were inhibited by the protein
phosphatase inhibitors okadaic acid and cantharidin. Dephosphorylated mutants of
claudin-1 and -2 were mainly distributed in the cytosol, which disappeared in
response to hypotonic stress. In contrast, mimicking phosphorylation mutants were
distributed in the TJs, which were not decreased by hypotonic stress. We suggest
that hypotonic stress induces dephosphorylation, clathrin-dependent endocytosis,
and degradation of claudin-1 and -2 in lysosomes, resulting in disruption of the
TJ barrier in renal tubular epithelial cells.