title: Radiation induced cytochrome c release causes loss of rat colonic fluid absorption by damage to crypts and pericryptal myofibroblasts.
abstract: BACKGROUND
  Therapeutic or accidental exposure to radiation commonly causes gastrointestinal disturbances, including diarrhoea. Rats subjected to whole body ionising radiation at a dose of 8 Gy lose their capacity to absorb fluid via the descending colon after four days. After seven days, fluid absorption recovers to control levels.
  
  
  OBJECTIVE
  To investigate the effect of ionising radiation on colonic permeability together with its effect on mitochondria dependent apoptotic signals and intercellular adhesion molecules.
  
  
  METHODS
  Rats were irradiated with doses of 0-12 Gy. Colonic permeability was measured by accumulation of fluorescein isothiocyanate (FITC) dextran in crypt lumens. Changes in levels of cytochrome c, caspase 3, E and OB cadherin, beta-catenin smooth muscle actin, and collagen IV were assessed using immunocytochemistry with confocal microscopy.
  
  
  RESULTS
  Cytosolic cytochrome c increased after 8 Gy (t(1/2) 1.4 (0.6) hours) and peaked at approximately six hours. Caspase 3 increased more slowly, particularly in crypt epithelial cells (t(1/2) 57 (14.5) hours). Pericryptal myofibroblasts disintegrated within 24 hours as was evident from loss of OB cadherin and smooth muscle actin. This coincided with increased crypt permeability to dextran. Intercellular adhesion between crypt luminal cells was not lost until day 4 when both beta-catenin and E-cadherin were minimal. The half maximal dose-response for these effects was in the range 2-4 Gy. Recovery of colonic transport was concurrent with recovery of pericryptal smooth muscle actin and OB cadherin. The pan caspase inhibitor Z-Val-Ala-Asp.fluoromethylketone (1 mg/kg per day) had a small effect in conserving the pericryptal sheath myofibroblasts and sheath permeability but had no systemic therapeutic effects.
  
  
  CONCLUSIONS
  These data suggest that radiation damage to the colon may be initiated by mitochondrial events. Loss of crypt fluid absorption and increased permeability coincided with decreased intercellular adhesion between crypt epithelial cells and loss of pericryptal sheath barrier function.
authors.affiliation: Institut de Protection et de Sûreté Nucléaire, Fontenay aux Roses, France.
authors.forename: J R
authors.initials: JR
authors.lastname: Thiagarajah
authors.forename: P
authors.initials: P
authors.lastname: Gourmelon
authors.forename: N M
authors.initials: NM
authors.lastname: Griffiths
authors.forename: F
authors.initials: F
authors.lastname: Lebrun
authors.forename: R J
authors.initials: RJ
authors.lastname: Naftalin
authors.forename: K C
authors.initials: KC
authors.lastname: Pedley
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism / Q000502:physiology; D002448:Cell Adhesion / Q000528:radiation effects; D003094:Collagen / Q000378:metabolism; D003106:Colon / Q000502:physiology / Q000528:radiation effects*; D003574:Cytochrome c Group / Q000378:metabolism / Q000528:radiation effects*; D003911:Dextrans / Q000493:pharmacokinetics; D004307:Dose-Response Relationship, Radiation; D005347:Fibroblasts / Q000378:metabolism / Q000528:radiation effects*; D016650:Fluorescein-5-isothiocyanate / Q000493:pharmacokinetics; D007408:Intestinal Absorption / Q000502:physiology / Q000528:radiation effects*; D008297:Male; D018613:Microscopy, Confocal; D051381:Rats; D017208:Rats, Wistar
chemical_list: D015820:Cadherins; D061945:Caspase Inhibitors; D003574:Cytochrome c Group; D003911:Dextrans; D003094:Collagen; D020169:Caspases; D016650:Fluorescein-5-isothiocyanate