Changes in the expression of claudins in active ulcerative colitis.
OBJECTIVE
Epithelial barrier function is impaired in ulcerative colitis (UC), but the pathophysiological mechanisms leading to this barrier defect are still far from clear. Because epithelial barrier function is primarily regulated by the most apical intercellular junction, referred to as the tight junction (TJ), we investigated the expression of TJ proteins on rectal epithelial mucosa in UC.


METHODS
Biopsies from the rectum of patients with active UC and of controls were studied. Tight-junction proteins were dual stained using specific antibodies (claudin-1, -2, -3, -4, and -7) as primary antibodies, and Cy-3 conjugated anti-rabbit IgG and Alexa488-conjugated anti-mouse IgG as secondary antibodies. Samples were analyzed by immunofluorescence microscopy. Immunoblotting and real-time polymerase chain reaction were performed to quantify TJ proteins and mRNA, respectively.


RESULTS
At TJ, claudin-4 and -7 staining was down-regulated in active UC, whereas claudin-2 staining was up-regulated. Claudin-4 and -7 proteins were down-regulated on immunoblotting, whereas claudin-2 protein was up-regulated in active UC. Claudin-1 and -3 expression levels were unchanged in controls and active UC. In active UC claudin-2 mRNA was increased, whereas claudin-4 and -7 mRNA were decreased.


CONCLUSIONS
Down-regulation of claudin-4 and claudin-7, and up-regulation of claudin-2, might lead to altered TJ structure and be related to the impaired epithelial function in active UC.
Oshima
Tadayuki
T
Department of Internal Medicine and Bioregulation, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan. t-oshima@hyo-med.ac.jp
Miwa
Hiroto
H
Joh
Takashi
T
D001706:Biopsy; D015153:Blotting, Western; D016022:Case-Control Studies; D062506:Claudin-4; D057167:Claudins; D003093:Colitis, Ulcerative / Q000378:metabolism*; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D008856:Microscopy, Fluorescence; D012333:RNA, Messenger / Q000032:analysis; D012007:Rectum / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019108:Tight Junctions / Q000737:chemistry*
C491875:CLDN2 protein, human; C568157:CLDN4 protein, human; C473255:CLDN7 protein, human; D062506:Claudin-4; D057167:Claudins; C568158:Cldn4 protein, mouse; D008565:Membrane Proteins; D012333:RNA, Messenger