Published ahead of print on October 27, 2005, doi:10.1164/rccm.200501-155OC
Am. J. Respir. Crit. Care Med., Volume 173, Number 3, February 2006, 334-344
A more recent version of this article appeared on February 1, 2006
Submitted on January 31, 2005
Accepted on October 21, 2005
Reactive Species Mediate Inhibition of Alveolar Type II Sodium Transport during Mycoplasma Infection
Judy M Hickman-Davis1, Carmel McNicholas-Bevensee2, Ian C Davis1, He-Ping Ma3, Glenda C Davis1, Charles A Bosworth1, and Sadis Matalon4*
1 Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, AL, USA,
2 Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA,
3 Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA,
4 Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Physiology and Biophysics, University of Alabama at Birmingham, Birmingham, AL, USA
* To whom correspondence should be addressed. E-mail: sadis{at}uab.edu.
Rationale: Mycoplasma pneumoniae is a significant cause of pneumonia in humans.
Objectives: Determine the impact of mycoplasma infection and the host inflammatory response on alveolar type II (ATII) cell ion transport.
Methods: Mice were infected with M. pulmonis for measurements of alveolar fluid clearance (AFC) in vivo and isolation of ATII cells. ATII cells were infected in vivo for determination of epithelial Na+ channel (ENaC) total and cell surface protein levels by biotinylation and Western blot, and in vitro for whole cell patch clamp recording and measurement of NO production by chemiluminescence.
Results: Mycoplasma infection significantly inhibited AFC at 24 hours and total and amiloride-sensitive AFC by 48 hours post infection (p.i.). In contrast, infected myeloperoxidase deficient mice had similar basal and amiloride-sensitive AFC values to uninfected controls at 48 hours p.i. Addition of forskolin restored total and amiloride-sensitive AFC to control values at 48 hours p.i. ATII cells isolated from infected mice demonstrated normal  , ,and  ENaC total protein levels; however, infected whole lung cell surface levels of ENaC were significantly decreased. Patch clamp recordings demonstrated a significant decrease in both total and amiloride sensitive Na+ currents at 24 hours p.i. ATII cells demonstrated a significant increase in the production of NO at 24 hours p.i., and inhibition of NO by ATII cells prior to infection reversed the decrease in total Na+ currents.
Conclusions: These data indicate that mycoplasma infection results in decreased AFC and functional ENaC via the production of reactive oxygen nitrogen intermediates.
Key words: epithelial sodium channels, amiloride, alveolar fluid clearance, patch clamp, chemiluminesce
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