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1 Cystic Fibrosis Research Laboratory, Stanford University, Stanford 94305-2130; and 2 Ethicon Endo-Surgery, Incorporated, Stanford, California 94305
We describe an optical method to quantify
single- gland secretion. Isolated tracheal mucosa were mounted at the
air-Krebs interface and coated with oil. Gland secretions formed
spherical bubbles that were digitally imaged at intervals, allowing
rates of secretion to be calculated. We monitored 340 glands in 54 experiments with 12 sheep. Glands secreted basally at low rates
(0.57 ± 0.04 nl · min
1 · gland1, 123 glands) in tissues up to 9 h postharvest and at lower rates for up
to 3 days. Carbachol (10 µM) stimulated secretion with an early
transient and a sustained or oscillating phase. Peak secretion was
15.7 ± 1.2 nl · min1 · gland1 (60 glands); sustained secretion was 4.5 ± 0.5 nl · min1 · gland1 (10 glands). Isoproterenol and phenylephrine (10 µM each) stimulated only
small, transient responses. We confirmed that cats have a large
secretory response to phenylephrine (11.6 ± 3.7 nl · min1 · gland1, 12 glands), but pigs, sheep, and humans all have small responses (<2
nl · min1 · gland1).
Carbachol-stimulated peak secretion was inhibited 56% by bumetanide, 67% by HCO
carbachol; phenylephrine; cystic fibrosis transmembrane conductance regulator; lung disease; mucociliary clearance
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