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J. Biol. Chem., Vol. 277, Issue 31, 28167-28175, August 2, 2002

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Mucus Secretion from Single Submucosal Glands of Pig
STIMULATION BY CARBACHOL AND VASOACTIVE INTESTINAL PEPTIDE^*

Nam Soo Joo, Yamil Saenz, Mauri E. Krouse, and Jeffrey J. Wine^§

From the Cystic Fibrosis Research Laboratory, Stanford University, Stanford, California 94305-2130 and  Ethicon Endo-Surgery, Inc., Stanford, California 94305

Secretion rates of >700 individual glands in isolated tracheal mucosa from 56 adult pigs were monitored optically. "Basal" secretion of 0.7 ± 0.1 nl·min¹ gland¹ was observed 1-9 h post-harvest but was near zero on day 2. Secretion to carbachol (10 µM) peaked at 2-3 min and then declined to a sustained phase. Peak secretion was 12.4 ± 1.1 nl·min¹ gland¹; sustained secretion was approximately one-third of peak secretion. Thapsigargin (1 µM) increased secretion from 0.1 ± 0.05 to 0.7 ± 0.2 nl·min¹ gland¹; thapsigargin did not cause contraction of the trachealis muscles. Isoproterenol and phenylephrine (10 µM each) were ineffective, but vasoactive intestinal peptide (1 µM) and forskolin (10 µM) each produced sustained secretion of 1.0 ± 0.5 and 1.7 ± 0.2 nl·min¹ gland¹, respectively. The density of actively secreting glands was 1.3/mm². Secretion to either carbachol or forskolin was inhibited (~50%) by either bumetanide or HCO removal and inhibited ~90% by the combined treatments. Mucus secreted in response to carbachol or forskolin was acidic by ~0.2 pH units relative to the bath and remained acidic by ~0.1 pH units after bumetanide. The strong secretory response to vasoactive intestinal peptide, the acidity of [cAMP]_i-stimulated mucus, and its inhibition by bumetanide were unexpected.

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