Inhaled ATP causes mucin release from goblet cells of intact rats.
Secretion of mucins from airway epithelial cells has been studied almost exclusively using in vitro cell culture systems. Our understanding of in vivo secretion is greatly limited due to the unavailability of both suitable model systems and adequate assays. It has been reported that ATP induces mucin release from the cultured primary tracheal surface epithelial cell, but there is no clear demonstration of the effect of ATP on mucin release in vivo, which is important to understand the mechanism of mucin release in vivo and also to devise means for regulation of mucin release. The objective of this experiment was to see if inhaled ATP could stimulate airway mucin release in intact rats using both enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. The results were: (1) a new monoclonal antibody (mAbRT03) developed against purified rat mucins specifically recognized high-molecular-mass mucins; (2) ELISA results with conventional gel-filtration assay results are virtually superimposable; (3) inhalation of ATP in intact rats resulted in a dose-independent increase in the amount of mucins in the tracheal lavage fluid with a concomitant decrease in the number of mucin-positive cells in the trachea. We conclude that extracellular ATP can stimulate mucin release from the airway in vivo, and the present rat inhalation system combined with ELISA of the airway secretions should serve a useful model for studying the pharmacology of airway mucin secretion in vivo.
Shin
C Y
CY
Laboratory of Pharmacology, College of Pharmacy, Seoul National University, Korea.
Kim
K C
KC
Lee
W J
WJ
Jo
M J
MJ
Park
K H
KH
Dalby
R
R
Ko
K H
KH
D000255:Adenosine Triphosphate / Q000008:administration & dosage / Q000494:pharmacology*; D000280:Administration, Inhalation; D000818:Animals; D000911:Antibodies, Monoclonal; D018929:Cell Culture Techniques; D002850:Chromatography, Gel / Q000379:methods; D017077:Culture Media, Conditioned / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D020397:Goblet Cells / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D009077:Mucins / Q000032:analysis / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014132:Trachea / Q000737:chemistry / Q000166:cytology
D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; D009077:Mucins; D000255:Adenosine Triphosphate