Changes in Lectin Binding by Differentiating Cutaneous Keratinocytes From the Newborn Rat

Surface glycoconjugates of cells from the basal layer of the skin of the newborn rat bind the isolectin I-B4 from Griffonia simplicifolia (GS I-B4) (alpha-D-galactosyl specificity). Surface glycoconjugates of the differentiated cells from the spinous and lower granular layers bind Ulex europeus agglutinin I (UEA) (alpha-L-fucosyl specificity). The change from GS I-B4 binding to UEA binding was studied in rat keratinocytes that were cultured as a monolayer in low-calcium medium until confluence, and then induced to stratify and terminally differentiate by raising the calcium concentration of the medium. The cells in the monolayer had basal cell morphology and exhibited surface binding of GS I-B4. However, at confluence, 30-40% of these cells also showed surface binding of UEA. There was an increase with time in the number of cells which bound both GS I-B4 and UEA. Raising the calcium concentration of the medium resulted in an increase in UEA binding. Cells of the upper layers of the stratifying cultures showed intense UEA binding but did not show any GS I-B4 binding. Double staining of frozen sections of newborn rat skin with fluorescein-conjugated GS I-B4 and rhodamine-conjugated UEA revealed that the surfaces of cells from the lower spinous layer bound both lectins. Thirty percent of the major glycoprotein fraction, that was isolated from the membranes of the epidermal cells of the newborn rat and was bound to an affinity column of UEA-Sepharose 4B, was also bound to an affinity column of GS I-B4-Sepharose 4B. These results indicate that surface glycoconjugates of rat keratinocytes differentiating in culture exhibit a change from GS I-B4 binding to UEA binding; the change in the cell surface glycoconjugates that results in the appearance of UEA binding, a feature of differentiated cells, occurs independently of stratification; and the change from GS I-B4 binding to UEA binding probably involves an "intermediate" glycoconjugate that binds both GS I-B4 and UEA and is found on the surface of cells from the lower spinous layer of the epidermis of the newborn rat.