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 zartilaginous 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 mug/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 mug/kg intravenously), a substance known to degranulate epithelial serous cells, reduced the staining of the serous cells for CGRP. CGRP-like immunoreactivity thus appears to be a useful marker of epithelial serous cells. We conclude that CGRP is present in the granules of epithelial serous cells in the rat trachea and is secreted into the tracheal lumen by stimuli that degranulate these cells.