Regulation of tight junction permeability by calcium mediators and cell cytoskeleton in rabbit tracheal epithelium.
The present study investigates the mechanisms controlling tight junction permeability of the tracheal epithelium, with an emphasis on the regulatory role of intra- and extracellular calcium as well as the cell cytoskeleton. The tracheas were isolated from rabbits and their junctional permeability barrier was investigated in vitro by means of transepithelial electrical resistance measurements and flux measurements of the radiolabeled paracellular tracer, 14C-mannitol. The effects of intra- and extracellular calcium were studied using the calcium ionophore A 23187 and EGTA, and that of the cytoskeleton was investigated using cytochalasin B. Intracellular calcium of the tracheal epithelium was monitored microfluorometrically using the specific calcium indicator, Fura-2 AM (acetoxymethyl ester). The results indicate that the tight junction permeability of the trachea was significantly increased upon treatment with all three of the test compounds, as evidenced by a substantial decrease in transepithelial electrical resistance and an increase in transepithelial flux of 14C-mannitol. The effects of EGTA and cytochalasin B on the tight junction permeability are fully reversible upon removal of the compounds from the bathing media. On the other hand, tissues treated with the calcium ionophore demonstrate a partial or no recovery in membrane permeability, depending on the intracellular calcium levels. Moderate and transient increases in intracellular calcium caused a partial reversibility of the membrane resistance, while high and sustained intracellular calcium levels induce a complete irreversibility of the membrane resistance.(ABSTRACT TRUNCATED AT 250 WORDS)
School of Pharmacy, West Virginia University, Morgantown 26506.
M
M
Bhat
D
D
Toledo-Velasquez
L
L
Wang
C J
CJ
Malanga
J K
JK
Ma
Y
Y
Rojanasakul
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology*; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects*; D004533:Egtazic Acid / Q000494:pharmacology; D004553:Electric Conductivity; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D016257:Fura-2; D066298:In Vitro Techniques; D008297:Male; D008353:Mannitol; D008856:Microscopy, Fluorescence; D010477:Perfusion; D010539:Permeability / Q000187:drug effects; D011817:Rabbits; D014132:Trachea / Q000187:drug effects / Q000378:metabolism*
D000001:Calcimycin; D003571:Cytochalasin B; D008353:Mannitol; D004533:Egtazic Acid; D002118:Calcium; D016257:Fura-2