Enhancement of Keratin Synthesis Induced by Lipokeratinogenoside, N-(O-Linoleoyl)-ω-Hydroxy Fatty Acyl Sphingosyl Glucose, in Association with Alteration of the Intracellular Ca2+-Content and Protein Kinase in Cultured Keratinocytes (FRSK)1

Abstract

Lipokeratinogenoside [N-(O-linoleoyl)-omega-hydroxy fatty acyl sphingosyl beta-glucose] is one of the epidermosides which were found to be glycosphingolipids characteristic of the epidermis of mammalian skin. On the addition of lipokeratinogenoside to cultured rat keratinocytes (FRSK), the amount of keratin in the cells increased, 48 and 144 h after cultivation, to 1.4 to 1.8 times higher than that without the addition of lipokeratinogenoside, and the number of cornified envelopes also significantly increased on cultivation of the cells with lipokeratinogenoside. Immunohistochemical staining with anti-keratin antibody revealed that the cells cultivated with lipokeratinogenoside were densely covered with keratin in distinct contrast to the control cells. The same enhanced syntheses of keratin and cornified envelopes were observed on cultivation in the presence of TPA, which has been shown to elevate the intracellular Ca(2+)-content and to translocate cytoplasmic protein kinase C to the plasma membrane in the initial stage of transmembrane signalling. Similarly, lipokeratinogenoside showed the ability to increase the intracellular Ca(2+)-content to the same extent as TPA did and to translocate protein kinase C to the membrane fraction. However, the above activities of lipokeratinogenoside decreased with removal of the linoleic acid moiety from lipokeratinogenoside with mild alkali, but linoleic acid alone did not show any activities, indicating that the lipokeratinogenoside molecule itself is required for expression of the activities.