There is temporal and spatial expression of alpha1 (IV), alpha2 (IV), alpha5 (IV), alpha6 (IV) collagen chains and beta1 integrins during the development of the basal lamina in an "in vitro" skin model.
Temporal and spatial expression of alpha1 (IV), alpha2 (IV), alpha3 (IV), alpha4 (IV), alpha5 (IV), and alpha6 (IV) collagen chains was studied during the formation of the basal lamina in an "in vitro" skin model. A sequential study was performed at 7-d and 14-d cultures (lamina densa absent) and at 28-, 36-, and 56-d cultures (lamina densa present). Expression of beta1, beta4, alpha1, alpha2, alpha3, alpha5, alpha6 integrin subunits and co-localization with collagen IV was studied by regular and laser confocal indirect immunofluorescence microscopy. mRNA expression of alpha2 (IV) and alpha6 (IV) chains was estimated by northern blots. The earliest expression of alpha1 (IV) and alpha2 (IV) collagen chains was noted in 7-d cultures restricted to basal keratinocytes. At 14-d cultures, alpha1 (IV) and alpha2 (IV) chains were noted in basal keratinocytes and as a broad band (10 microm) in the adjacent dermis. At this stage 80% of the alpha2 (IV) mRNA was expressed in the dermis and 20% in the epidermis. At 28-, 36-, and 56-d cultures the alpha1 (IV) and alpha2 (IV) chains were present in a linear distribution at the epidermo-dermal junction and in the upper dermis. The alpha6 (IV) collagen chains were expressed much later at 36-d cultures and the alpha5 (IV) at 56 d, both mostly in a linear distribution but also in the adjacent dermis. Alpha6 (IV) mRNA was demonstrated in the dermis of 36-d cultures. There was co-localization of collagen IV and beta1 integrin subunits in 14-d cultures at the matrix site of keratinocytes. Functional perturbation studies with AIIB2 monoclonal antibody (anti-beta1 subunits) and competitive inhibition with a collagen cyanogen bromide digestion derived fragment (CB3[IV]) that contains the collagen IV ligand for alpha1beta1, alpha2beta1 integrins, altered the pattern of collagen IV deposition.
Fleischmajer
R
R
Department of Dermatology, Mount Sinai School of Medicine, New York, NY 10029, U.S.A.
Kühn
K
K
Sato
Y
Y
MacDonald
E D
ED
Perlish
J S
JS
Pan
T C
TC
Chu
M L
ML
Kishiro
Y
Y
Oohashi
T
T
Bernier
S M
SM
Yamada
Y
Y
Ninomiya
Y
Y
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001485:Basement Membrane / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D018920:Coculture Techniques; D003094:Collagen / Q000737:chemistry / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism*; D015603:Keratinocytes / Q000166:cytology; D008854:Microscopy, Electron; D010446:Peptide Fragments / Q000494:pharmacology; D012867:Skin / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism*; D013997:Time Factors; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D019012:Integrin beta1; D010446:Peptide Fragments; D003094:Collagen