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
| Autor: | Raul Fleischmajer, Te Cheng Pan, Yoshifumi Ninomiya, E. Douglas MacDonald, Yoshikazu Sato, Jerome S. Perlish, Y Yamada, Mon-Li Chu, Klaus Kühn, Yumiko Kishiro, Toshitaka Oohashi, Suzanne M. Bernier |
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| Rok vydání: | 1997 |
| Předmět: |
Keratinocytes
Time Factors Integrin Dermatology Biology Biochemistry Basement Membrane chemistry.chemical_compound Dermis collagen IV medicine Humans Tissue Distribution Northern blot Molecular Biology Skin Basement membrane urogenital system Integrin beta1 Antibodies Monoclonal Cell Biology Fibroblasts Molecular biology Coculture Techniques Peptide Fragments Microscopy Electron medicine.anatomical_structure chemistry Cell culture biology.protein Basal lamina Lamina densa Cyanogen bromide Collagen |
| Zdroj: | The Journal of investigative dermatology. 109(4) |
| ISSN: | 0022-202X |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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