Reestablishment of a mucociliary epithelium in tracheal organ cultures exposed to retinyl acetate: a biochemical and morphometric study.
Tracheal explants derived from vitamin A-deficient rats underwent keratinizine squamous metaplasia in organ culture when grown in serum-free medium. Within 1 d after the addition of 0.1, 2, or 10 microgram retinyl acetate per ml of medium, there was a concentration-dependent increase in the uptake of [3H]glucosamine and [14C]serine into both the total mucous glycoprotein and the principal purified mucin fraction eluted from a DEAE-Sephacel column with 0.2 M NaCl. The stimulation of mucin synthesis continued throughout the 21-d exposure period in a concentration-dependent fashion. It was also found that vitamin A had a greater effect on the incorporation of [3H]glucosamine than on [14C]serine into the secreted mucins, particularly at the higher retinyl acetate concentrations. This result indicated a greater effect of the vitamin on the synthesis of the carbohydrate moiety of the mucins. Morphological analysis by light and electron microscopy demonstrated that the keratinizing squamous epithelium began to revert to a mucus-secreting tissue as early as 24 h after addition of 10 microgram retinyl acetate to the medium. The response was slower with the lower vitamin concentrations. Stereological analysis revealed that the increase in the volume fraction of the Golgi apparatus reached a stable level which could not be altered with continued exposure to retinyl acetate, but that the volume fraction of mucin droplets continually increased and apparently did not reach a maximum in the 21-d exposure period. Conversely, the volume fraction of filament bundles and the number of desmosomes decreased during the vitamin A treatment.
D000818:Animals; D002478:Cells, Cultured; D002923:Cilia / Q000648:ultrastructure*; D003470:Culture Media; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D008854:Microscopy, Electron; D009092:Mucous Membrane / Q000648:ultrastructure*; D051381:Rats; D011916:Rats, Inbred F344; D013047:Specific Pathogen-Free Organisms; D014132:Trachea / Q000378:metabolism / Q000648:ultrastructure*; D014801:Vitamin A / Q000494:pharmacology*
D003470:Culture Media; D006023:Glycoproteins; D014801:Vitamin A