Increased acetylcholine release in tracheas from allergen-exposed IgE-immune mice

G. L. Larsen, T. M. Fame, H. Renz, J. E. Loader, J. Graves, M. Hill and E. W. Gelfand Department of Pediatrics, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado. Increased release of acetylcholine (ACh) from airway parasympathetic nerve endings is one mechanism that may contribute to increases in airway responsiveness in immunoglobulin E (IgE)-immune allergen-exposed animals. We measured ACh released from murine tracheas following electrical field stimulation in vitro. BALB/c mice were immunized by exposure to an aerosol of 1% ovalbumin in sterile phosphate-buffered saline for 20 min/day for 10 days. At this time, levels of ovalbumin-specific IgE were proportionately higher than ovalbumin-specific IgG. As a control, nonimmune mice were similarly exposed to phosphate-buffered saline alone. Forty-eight hours after the last aerosol, tracheas were removed for assessment of either the contractile responses to electrical field stimulation and a cholinergic agonist (methacholine or ACh) or release of ACh produced by electrical field stimulation. ACh in the bath was measured using high-performance liquid chromatography with electrochemical detection. The stimulation frequencies causing one-half the maximal contractile response to electrical field stimulation were 4.1 +/- 0.2 and 2.8 +/- 0.2 Hz (P = 0.0001) for nonimmune and immune mice, respectively, whereas the molar concentrations of methacholine causing one-half of the maximal contractile response did not significantly differ. In addition, the dose-response curves of immune and nonimmune tracheas to ACh were superimposable. A significant increase in ACh release was demonstrated at both 10 and 20 Hz in tracheas from immune mice. ACh release (pmol.g tissue-1.min-1) from nonimmune and immune murine tracheas, respectively, were 140 +/- 8 and 205 +/- 22 (P = 0.013) at 10 Hz and 147 +/- 13 and 227 +/- 14 (P = 0.008) at 20 Hz.