title: Initiation of skin basement membrane formation at the epidermo-dermal interface involves assembly of laminins through binding to cell membrane receptors.
abstract: To study the mechanism of basement membrane formation, we determined by immunochemistry temporal and spatial expression of laminin-5 (Ln-5), laminin-1 (Ln-1) and their integrin receptors during early skin morphogenesis. A 3-dimensional skin culture was used that allows the study of the sequential molecular events of basement membrane formation at the epidermodermal interface. During early anchorage of keratinocytes to the extracellular matrix there is expression of Ln-5, BP-230 antigen and alpha3, beta1 integrin subunits. During epidermal stratification and prior to the formation of the lamina densa there is assembly of Ln-5, Ln-1, collagen IV and nidogen accompanied by keratinocyte basal clustering of alpha2, alpha3, alpha6, beta1, and beta4+ integrin subunits. The assembly pattern of Ln-1 and Ln-5 can be disturbed with functional antibodies against the beta1 (AIIB2) and alpha6 (GoH3) integrin subunits. Ln-1 assembly can also be disturbed with antibodies against its E8 domain and by competitive inhibition with a synthetic peptide (AG-73) derived from its G-4 domain. Quantitative RT-PCR showed that the dermis contributes about 80% of the laminin gamma)1 chain mRNA while 20% is produced by the epidermis which emphasizes its dual tissue origin and the major contribution of the mesenchyma in laminin production. The laminin gamma2 chain mRNA, present in Ln-5, was mostly of epidermal origin. This study presents evidence that during the initiation of basement membrane formation, laminins bind to keratinocyte plasma membrane receptors and thus may serve as nucleation sites for further polymerization of these compounds by a self-assembly process.
authors.affiliation: Department of Dermatology, Mount Sinai School of Medicine, New York, NY 10029, USA.
authors.forename: R
authors.initials: R
authors.lastname: Fleischmajer
authors.forename: A
authors.initials: A
authors.lastname: Utani
authors.forename: E D
authors.initials: ED
authors.lastname: MacDonald
authors.forename: J S
authors.initials: JS
authors.lastname: Perlish
authors.forename: T C
authors.initials: TC
authors.lastname: Pan
authors.forename: M L
authors.initials: ML
authors.lastname: Chu
authors.forename: M
authors.initials: M
authors.lastname: Nomizu
authors.forename: Y
authors.initials: Y
authors.lastname: Ninomiya
authors.forename: Y
authors.initials: Y
authors.lastname: Yamada
mesh_terms: D008841:Actin Cytoskeleton / Q000502:physiology / Q000648:ultrastructure; D000595:Amino Acid Sequence; D001485:Basement Membrane / Q000166:cytology / Q000502:physiology*; D002450:Cell Communication; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003896:Desmosomes / Q000502:physiology / Q000648:ultrastructure; D000078404:Epidermal Cells; D004817:Epidermis / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D006801:Humans; D007150:Immunohistochemistry; D019012:Integrin beta1 / Q000096:biosynthesis; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology*; D007797:Laminin / Q000096:biosynthesis* / Q000138:chemical synthesis / Q000378:metabolism; D008297:Male; D009024:Morphogenesis; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000378:metabolism; D012867:Skin / Q000166:cytology*; D012879:Skin Physiological Phenomena
chemical_list: C105786:AG 73; D019012:Integrin beta1; D007797:Laminin; D010446:Peptide Fragments