Effect of retinoic acid on cornified envelope formation: difference between spontaneous envelope formation in vivo or in vitro and expression of envelope competence.
A large number of cross-linked envelopes form spontaneously when cell lines derived from chemically induced mouse skin papillomas are cultured in medium containing 1.2 mM calcium. This phenomenon is associated with high activity of the cross-linking enzyme, epidermal transglutaminase (TGase). The influence of retinoic acid (RA) on envelope formation was studied in detail in a papilloma cell line, PE. Retinoic acid (3 microM) completely blocked cornified envelope (CE) production but reduced TGase activity only 50%. A rabbit antiserum was produced against sonicated CEs isolated from newborn mouse skin. On Western blots of epidermal extracts, diffuse staining was observed for particulate proteins of suprabasal, but not basal, cells and similar immunoreactive material was absent from the cytosolic fraction of both cell layers. The antibody also recognized particulate proteins from PE cells induced to differentiate by calcium, but not from cells grown in the presence of high calcium and RA. The antiserum appears to recognize partially cross-linked CE precursor proteins judging by the diffuse staining, the molecular weight range of the proteins stained, and their origin in the particulate cellular fraction. Cross-linked envelopes could be induced in RA-treated PE cells by permeabilization with 0.75 M NaCl or 50 micrograms/ml A23187. However, this treatment failed to cause the appearance of proteins recognized by the antiserum. Preincubation of the antiserum with purified fragments of CEs from newborn mouse epidermis, but not with cross-linked envelopes from permeabilized, RA-treated PE cells, removed immunoreactivity. These results indicate that the cross-linked envelopes formed in RA-treated cells after permeabilization lack a set of proteins contained in CEs from stratum corneum and may even be composed of different proteins. Retinoic acid appears to prevent CE formation in part by inhibiting activation of epidermal TGase but in addition by influencing the synthesis of precursor proteins.
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D000078404:Epidermal Cells; D004817:Epidermis / Q000187:drug effects* / Q000201:enzymology / Q000648:ultrastructure; D007158:Immunologic Techniques; D010212:Papilloma / Q000201:enzymology / Q000648:ultrastructure; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors; D011503:Transglutaminases / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology*
D000001:Calcimycin; D012965:Sodium Chloride; D014212:Tretinoin; D011503:Transglutaminases