title: Preserved Na(+)/H(+) exchanger isoform 3 expression and localization, but decreased NHE3 function indicate regulatory sodium transport defect in ulcerative colitis.
abstract: BACKGROUND
  A major causative factor of diarrhea in ulcerative colitis (UC) patients is the loss of Na(+) absorptive capacity of the inflamed colonic mucosa. Potential contributing mechanisms include reduced driving force for active transport, and impaired expression, mislocalization, or defective transport function of Na(+) absorptive proteins. We therefore studied the expression, brush border membrane (BBM) localization, and transport capacity of the major intestinal Na(+) absorptive protein, the Na(+)/H(+) exchanger isoform 3 (NHE3) in biopsies from UC patients.
  
  
  METHODS
  In UC and control biopsies, inflammation was graded histologically, NHE3, tumor necrosis factor alpha (TNF-alpha), villin, as well as other housekeeping genes were analyzed by quantitative real-time polymerase chain reaction (PCR), BBM localization of NHE3 determined by immunohistochemistry, and confocal microscopy. Na(+) absorptive capacity was assessed by (22)Na(+) isotope fluxes and NHE3 transport activity measured microfluorometrically in BCECF-loaded surface colonocytes within isolated crypts.
  
  
  RESULTS
  In mildly, moderately, and severely inflamed sigmoid colon of UC patients, neither NHE3 mRNA expression nor the abundance of NHE3 in the BBM was significantly altered compared to other structural components of the BBM. However, Na(+) absorption was strongly reduced by approximately 80% and acid-activated NHE3 transport activity was significantly decreased in the surface cells of sigmoid colonic crypts even in moderately inflamed mucosa.
  
  
  CONCLUSIONS
  In the colonic mucosa of patients with active UC, NHE3 transport capacity was found significantly decreased despite correct NHE3 location and abundance in the brush border, independent of current treatment. These findings suggest functional NHE3 transport as a novel factor for inflammatory diarrhea in UC patients.
authors.affiliation: Department of Gastroenterology, Hannover Medical School, Hannover, Germany.
authors.forename: Sunil
authors.initials: S
authors.lastname: Yeruva
authors.forename: Klaudia
authors.initials: K
authors.lastname: Farkas
authors.forename: Jessica
authors.initials: J
authors.lastname: Hubricht
authors.forename: Katja
authors.initials: K
authors.lastname: Rode
authors.forename: Brigitte
authors.initials: B
authors.lastname: Riederer
authors.forename: Oliver
authors.initials: O
authors.lastname: Bachmann
authors.forename: Ayhan
authors.initials: A
authors.lastname: Cinar
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Rakonczay
authors.forename: Tamás
authors.initials: T
authors.lastname: Molnár
authors.forename: Ferenc
authors.initials: F
authors.lastname: Nagy
authors.forename: Jochen
authors.initials: J
authors.lastname: Wedemeyer
authors.forename: Michael
authors.initials: M
authors.lastname: Manns
authors.forename: Dirk
authors.initials: D
authors.lastname: Raddatz
authors.forename: Mark W
authors.initials: MW
authors.lastname: Musch
authors.forename: Eugene B
authors.initials: EB
authors.lastname: Chang
authors.forename: Péter
authors.initials: P
authors.lastname: Hegyi
authors.forename: Ursula
authors.initials: U
authors.lastname: Seidler
mesh_terms: D015153:Blotting, Western; D003093:Colitis, Ulcerative / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003106:Colon / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007124:Immunoenzyme Techniques; D007408:Intestinal Absorption; D017136:Ion Transport; D008871:Microvilli / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012964:Sodium / Q000378:metabolism*; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; C000613385:SLC9A3 protein, human; D000074083:Sodium-Hydrogen Exchanger 3; D017923:Sodium-Hydrogen Exchangers; D014409:Tumor Necrosis Factor-alpha; D012964:Sodium