Reconstitution of the epidermal basement membrane after enzymatic dermal-epidermal separation of embryonic mouse skin.
Pieces of trypsin-isolated 14-day embryonic mouse epidermis were recombined with various living or non-living dermal or non-dermal substrates, in order to analyse the reconstruction of the dermal-epidermal junction. The constitution and ultrastructure of the epidermal basement membrane were characterized by immunolabelling of laminin, type IV collagen and bullous pemphigoid antigen, and by transmission electron microscopy. Trypsin treatment of dorsal skin followed by dermal-epidermal separation does not visibly damage the epidermal basement membrane, which remains attached to the lower face of epidermis. When freshly isolated epidermis is reassociated with dermis, the basement membrane is first degraded during the first 4 h of culture, then reconstituted within 24 h. When epidermis is cultured in isolation the basement membrane disappears within 4 h and is not reconstructed. Epidermis, precultured for 4 h and thus deprived of its basement membrane prior to reassociation, is able to reconstruct an antigenically and ultrastructurally normal basement membrane, when recombined with living or frozen-killed (-20 degrees C) dermis, with muscle tissue, or with a film of fibrous type I collagen. No basement membrane is reconstituted when the epidermis is recombined with heat (100 degrees C) killed dermis. It is concluded that, in the reconstituted epidermal basement membrane, laminin, type IV collagen, bullous pemphigoid antigen, and lamina densa are of exclusive epidermal origin.
D000818:Animals; D001485:Basement Membrane / Q000166:cytology* / Q000648:ultrastructure; D002469:Cell Separation; D000078404:Epidermal Cells; D004817:Epidermis / Q000648:ultrastructure; D006358:Hot Temperature; D051379:Mice; D008854:Microscopy, Electron; D009924:Organ Culture Techniques; D012867:Skin / Q000166:cytology* / Q000196:embryology / Q000648:ultrastructure; D014357:Trypsin
D014357:Trypsin