Expression of “Cell-type-specific” Markers during Rat Tracheal Epithelial Regeneration

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Abstract
Previous studies showed that Griffonia simplicifolia I-isolectin B4 (GS I-B4) and monoclonal antibodies (mAbs) against keratin 14 labeled basal cells in the adult rat trachea while other mAbs specifically stained secretory and/or ciliated cells. We used these "cell-type-specific" markers to study cellular differentiation during tracheal epithelial regeneration. Denuded tracheal grafts were inoculated with rat tracheal epithelial cells and were implanted in syngeneic hosts. Marker expression was correlated with the appearance of morphologically defined cell types. At 4 days, the epithelium was squamoid, one to three cell layers thick, and was apparently composed of a single morphologic cell type. Because this cell did not exhibit distinguishing features of any mature tracheal cell, we provisionally termed it the "poorly differentiated cell" (PD cell). PD cells expressed keratin 14 and GS I-B4 binding sites; they contained glycogen and had lipid droplets but did not react with secretory or ciliated cell-specific mAbs. At 7 days, areas of the epithelium were pseudostratified and secretory cell-specific markers were present at the apex of differentiating columnar cells; ultrastructurally, these cells resembled secretory cells in adult tracheas. Simultaneously, a few preciliated and ciliated cells appeared that expressed a ciliated cell-specific epitope. No cells were observed coexpressing secretory and ciliated cell markers. Basal cells also became recognizable on day 7. These expressed keratin 14 and GS I-B4 binding sites throughout the study. Newly appearing secretory and ciliated cells also expressed these two markers initially but lost them gradually as the mucociliary epithelium matured. In the tracheal graft model of epithelial regeneration, the PD cells were pivotal intermediates from which all differentiated cells developed. Basal cells continued to express the same markers as PD cells, which were gradually lost in secretory and ciliated cells as they acquired new sets of specific epitopes.
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