Calcitonin gene-related peptide in secretory granules of serous cells in the rat tracheal epithelium.
The tracheal epithelium of pathogen-free rats consists mainly of serous-type secretory cells, ciliated cells, basal cells, and a few neuroendocrine cells. Mucus-containing goblet cells are rare. Calcitonin gene-related peptide (CGRP) is known to exist in the neuroendocrine cells and in sensory nerves of the tracheal mucosa and is released into the airway lumen by sensory nerve stimulation. In this study, we determined whether epithelial serous cells are another source of CGRP. Tracheas of adult male specific pathogen-free F344 rats were immunostained by an avidin-biotin technique either as whole mounts or as cryostat sections using two different polyclonal primary antibodies to rat CGRP. Some specimens were stained for CGRP-like immunofluorescence and examined with a confocal microscope. CGRP immunoreactivity was present in granules of serous cells throughout the trachea. In whole mounts, the stained cells were most abundant between the cartilaginous rings, especially in the rostral trachea, where they constituted 56% of the epithelial cells in contact with the tracheal lumen. Serous cells were easily distinguished from neuroendocrine cells and nerve fibers with CGRP immunoreactivity. In evidence that the CGRP immunoreactivity was specific, the staining of serous cells was abolished by omitting the primary antibody and by absorption with 10 micrograms/ml CGRP. Antibodies to substance P, vasoactive intestinal polypeptide, and tyrosine hydroxylase did not stain epithelial serous cells. An antibody to protein gene product 9.5 labeled neuroendocrine cells, but not serous cells. Injection of capsaicin (150 micrograms/kg intravenously), a substance known to degranulate epithelial serous cells, reduced the staining of the serous cells for CGRP.(ABSTRACT TRUNCATED AT 250 WORDS)
Baluk
P
P
Cardiovascular Research Institute, University of California, San Francisco 94143-0130.
Nadel
J A
JA
McDonald
D M
DM
D000704:Analysis of Variance; D000818:Animals; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis*; D015550:Cell Degranulation; D003594:Cytoplasmic Granules / Q000737:chemistry*; D004847:Epithelial Cells; D004848:Epithelium / Q000737:chemistry; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D013373:Substance P / Q000032:analysis; D013869:Thiolester Hydrolases / Q000032:analysis; D014132:Trachea / Q000737:chemistry* / Q000166:cytology; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis; D043222:Ubiquitin Thiolesterase; D014660:Vasoactive Intestinal Peptide / Q000032:analysis
D013373:Substance P; D014660:Vasoactive Intestinal Peptide; D014446:Tyrosine 3-Monooxygenase; D013869:Thiolester Hydrolases; D043222:Ubiquitin Thiolesterase; D015740:Calcitonin Gene-Related Peptide