Carbohydrate cytochemistry of tracheobronchial airway epithelium of the rabbit

Three types of nonciliated secretory epithelial cells contribute to the mucous lining of pulmonary airways: mucous cells, serous cells, Clara cells. Contrary to observations in other species, airways to the rabbit have very few mucous cells. In the rabbit, the predominant secretory cell through the entire airway tree, including the trachea, appears to be one cell type, Clara cell. While these cells share the same ultrastructural features throughout the tree, the nature of their contribution to the mucous blanket is not clear. This study was designed to characterize the carbohydrate components of secretory granules in tracheal Clara cells, and to compare that carbohydrate with that of tracheal mucous (goblet) cells and with Clara cells of more distal airway generations. Trachea and lungs of six adult male rabbits were fixed by airway infusion, the conducting airways of the right cranial lobe dissected and tissue selected from the trachea and five dorsal airway generations. For light microscopy (LM), sections of paraffin-embedded tissues were stained with Alcian blue-periodic-acid-Schiff (AB/PAS), dialyzed iron (DI), and high iron diamine-Alcian blue (HID-AB). For electron microscopy (EM), fixed tissues were incubated with DI, HID, MgCl₂ or buffer, postosmicated, embedded in epoxy resin, and thin sections stained with periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP). By LM, most Clara cells did not react with PAS, AB HID, or DI. A few in trachea and bronchi had PAS-positive apical margins. Mucous goblet cells were positive with PAS, AB, and HID, indicating sulfated glycoproteins. By EM, a small number of Clara cells had PA-TCH-SP-positive luminal granules, a few luminal granules had DI-positive rims. Almost all Clara cell granules were negative with PA-TCH-SP, HID, and DI. The granules of mucous goblet cells had a finely granular core surrounded by a meshwork of variable density. The meshwork was positive wih PA-TCH-SP, DI, and HID. The cores were not. We concluded that: 1) the Clara cell does not contribute carbohydrates to the airway mucous lining: 2) mucous goblet cells secrete predominantly sulfated glycoproteins; and 3) the contribution to mucous carbohydrates by Clara cells does not vary with the airway level in which they are located.