Epithelial transport and barrier function in occludin-deficient mice.
OBJECTIVE
This study aimed at functional characterization of the tight junction protein occludin using the occludin-deficient mouse model.


METHODS
Epithelial transport and barrier functions were characterized in Ussing chambers. Impedance analysis revealed the ionic permeability of the epithelium (Re, epithelial resistance). Conductance scanning differentiated transcellular (Gc) and tight junctional conductance (Gtj). The pH-stat technique quantified gastric acid secretion.


RESULTS
In occludin+/+ mice, Re was 23+/-5 Omega cm2 in jejunum, 66+/-5 Omega cm2 in distal colon and 33+/-6 Omega cm2 in gastric corpus and was not altered in heterozygotic occludin+/- or homozygotic occludin-/- mice. Additionally, [3H]mannitol fluxes were unaltered. In the control colon, Gc and Gtj were 7.6+/-1.0 and 0.3+/-0.1 mS/cm2 and not different in occludin deficiency. Epithelial resistance after mechanical perturbation or EGTA exposition (low calcium switch) was not more affected in occludin-/- mice than in control. Barrier function was measured in the urinary bladder, a tight epithelium, and in the stomach. Control Rt was 5.8+/-0.8 kOmega cm2 in urinary bladder and 33+/-6 Omega cm2 in stomach and not altered in occludin-/- mice. In gastric corpus mucosa, the glandular structure exhibited a complete loss of parietal cells and mucus cell hyperplasia, as a result of which acid secretion was virtually abolished in occludin-/- mice.


CONCLUSIONS
Epithelial barrier characterization in occludin-deficiency points against an essential barrier function of occludin within the tight junction strands or to a substitutional redundancy of single tight junction molecules like occludin. A dramatic change in gastric morphology and secretory function indicates that occludin is involved in gastric epithelial differentiation.
Schulzke
J D
JD
Department of Gastroenterology, Campus Benjamin Franklin, Charité-Universitary Medicine Berlin, Germany. joerg.schulzke@charite.de
Gitter
A H
AH
Mankertz
J
J
Spiegel
S
S
Seidler
U
U
Amasheh
S
S
Saitou
M
M
Tsukita
S
S
Fromm
M
M
D000818:Animals; D003106:Colon / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism*; D006579:Heterozygote; D006720:Homozygote; D015151:Immunoblotting; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D062793:Occludin; D016133:Polymerase Chain Reaction; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D019108:Tight Junctions / Q000378:metabolism*; D001743:Urinary Bladder / Q000378:metabolism
D008565:Membrane Proteins; D062793:Occludin; C568986:Ocln protein, mouse