Airway surface liquid pH in well-differentiated airway epithelial cell cultures and mouse trachea

Author:

Jayaraman Sujatha1,Song Yuanlin1,Verkman A. S.1

Affiliation:

1. Departments of Medicine and Physiology, Cardiovascular Research Institute, University of California, San Francisco, California 94143-0521

Abstract

Airway surface liquid (ASL) pH has been proposed to be important in the pathophysiology of cystic fibrosis, asthma, and cough. Ratio image analysis was used to measure pH in the ASL after staining with the fluorescent pH indicator 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF)-dextran. ASL pH in bovine airway cell cultures grown at an air-liquid interface was 6.98 ± 0.06 in the absence and 6.81 ± 0.04 in the presence of HCO[Formula: see text]/CO2. Steady-state ASL pH changed in parallel to changes in bath pH and was acidified by Na+ or Cl replacement but was not affected by the inhibitors amiloride, glibenclamide, or 4,4′-dinitrostilbene-2,2′-disulfonic acid. In response to sudden acidification or alkalization of the ASL by ∼0.4 pH units by HCl/NaOH, ASL pH recovered to its initial value at a rate of 0.035 pH units/min (−HCO[Formula: see text]) and 0.060 pH units/min (+HCO[Formula: see text]); the pH recovery rate was reduced by amiloride and H2DIDS. In anesthetized mice in which the trachea was surgically exposed for measurement of BCECF-dextran fluorescence through the translucent tracheal wall, ASL pH was 7.14 ± 0.01. ASL pH was sensitive to changes in blood pH created by metabolic (HCl or NaHCO3 infusion) or respiratory (hyperventilation, hypoventilation) mechanisms. ASL pH is thus primarily determined by basolateral fluid pH, and H+/OH transport between the ASL and basolateral fluid involves amiloride-sensitive Na+/H+ exchange and stilbene-sensitive Cl/HCO[Formula: see text] exchange. The rapid response of ASL pH to changes in systemic acid-base status may contribute to airway hypersensitivity in asthma and other airway diseases.

Publisher

American Physiological Society

Subject

Cell Biology,Physiology

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