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Am J Physiol Cell Physiol 275: C1284-C1290, 1998;
0363-6143/98 $5.00
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Vol. 275, Issue 5, C1284-C1290, November 1998

Pseudomonas aeruginosa induces changes in fluid transport across airway surface epithelia

D. J. Evans1, P. S. Matsumoto2, J. H. Widdicombe2,3, C. Li-Yun1, A. A. Maminishkis1, and S. S. Miller1,4

1 School of Optometry and 4 Department of Molecular and Cell Biology, University of California, Berkeley 94720; 2 Children's Hospital Oakland Research Institute, Oakland 94609; and 3 Department of Physiology and Cardiovascular Research Institute, University of California, San Francisco, California 94143

Fluid transport across cultures of bovine tracheal epithelium was measured with a capacitance probe technique. Baseline fluid absorption (Jv) across bovine cells of 3.2 µl · cm-2 · h-1 was inhibited by ~78% after 1 h of exposure to suspensions of Pseudomonas aeruginosa, with a concomitant decrease in transepithelial potential (TEP) and increase in transepithelial resistance (Rt). Effects of P. aeruginosa were blocked by amiloride, which decreased Jv by 112% from baseline of 2.35 ± 1.25 µl · cm-2 · h-1, increased Rt by 101% from baseline of 610 ± 257 Omega  · cm2, and decreased TEP by 91% from baseline of -55 ± 18.5 mV. Microelectrode studies suggested that effects of P. aeruginosa on amiloride-sensitive Na absorption were due in part to a block of basolateral membrane K channels. In the presence of Cl transport inhibitors [5-nitro-2-(3-phenylpropylamino)-benzoic acid, H2-DIDS, and bumetanide], P. aeruginosa induced a fluid secretion of ~2.5 ± 0.4 µl · cm-2 · h-1 and decreased Rt without changing TEP. However, these changes were abolished when the transport inhibitors were used in a medium in which Cl was replaced by an impermeant organic anion. Filtrates of P. aeruginosa suspensions had no effect on Jv, TEP, or Rt. Mutants lacking exotoxin A or rhamnolipids or with defective lipopolysaccharide still inhibited fluid absorption and altered bioelectrical properties. By contrast, mutations in the rpoN gene encoding a sigma  factor of RNA polymerase abolished actions of P. aeruginosa. In vivo, changes in transepithelial salt and water transport induced by P. aeruginosa may alter viscosity and ionic composition of airway secretions so as to foster further bacterial colonization.

bacteria; RpoN


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