Normal mucus formation requires cAMP-dependent HCO3− secretion and Ca2+-mediated mucin exocytosis

摘要

Evidence from the pathology in cystic fibrosis (CF) and recent results in vitro indicate that HCO3⁻ is required for gel-forming mucins to form the mucus that protects epithelial surfaces. Mucus formation and release is a complex process that begins with an initial intracellular phase of synthesis, packaging and apical granule exocytosis that is followed by an extracellular phase of mucin swelling, transport and discharge into a lumen. Exactly where HCO3⁻ becomes crucial in these processes is unknown, but we observed that in the presence of HCO3⁻, stimulating dissected segments of native mouse intestine with 5-hydroxytryptamine (5–HT) and prostaglandin E2 (PGE2) induced goblet cell exocytosis followed by normal mucin discharge in wild-type (WT) intestines. CF intestines that inherently lack cystic fibrosis transmembrane conductance regulator (CFTR)-dependent HCO3⁻ secretion also demonstrated apparently normal goblet cell exocytosis, but in contrast, this was not followed by similar mucin discharge. Moreover, we found that even in the presence of HCO3⁻, when WT intestines were stimulated only with a Ca²⁺-mediated agonist (carbachol), exocytosis was followed by poor discharge as with CF intestines. However, when the Ca²⁺-mediated agonist was combined with a cAMP-mediated agonist (isoproterenol (isoprenaline) or vasoactive intestinal peptide) in the presence of HCO3⁻ both normal exocytosis and normal discharge was observed. These results indicate that normal mucus formation requires concurrent activation of a Ca²⁺-mediated exocytosis of mucin granules and an independent cAMP-mediated, CFTR-dependent, HCO3⁻ secretion that appears to mainly enhance the extracellular phases of mucus excretion.