title: Targeted inactivation of murine laminin gamma2-chain gene recapitulates human junctional epidermolysis bullosa.
abstract: Junctional forms of epidermolysis bullosa (JEB) are associated with mutations in six distinct genes expressed in the cutaneous basement membrane zone; these include LAMA3, LAMB3, and LAMC2, which encode laminin 5 subunit polypeptides, the alpha3-, beta3-, and gamma2-chains, respectively. Here we generated a mouse model for JEB by inactivating the laminin gamma2-chain gene by targeted frameshift deletion of exon 8 in Lamc2. Heterozygous mice were phenotypically normal, whereas the majority of Lamc2-/- mice showed blistering phenotype on days 1 to 2 and died within 5 days of birth. The Lamc2-/- mice demonstrated absent expression of laminin gamma2-chain on the basement membrane zone as well as attenuated expression of alpha3- and beta3-chains of laminin. Transmission electron microscopy revealed rudimentary, poorly developed hemidesmosomes. The epidermis of the Lamc2-/- mice revealed induced apoptosis in the basal cells of the blistered skin, suggesting that cell-matrix adhesion provided by laminin 5 plays a role in cell survival in vivo. Cultured Lamc2-/- keratinocytes demonstrated slightly positive staining with gamma2-chain-specific antibodies, which could be explained by the presence of a transcript with partial restoration of the reading frame owing to alternative splicing in vitro. These cells proliferated in different matrices and attached to type IV collagen and Matrigel as efficiently as the wild-type keratinocytes, whereas their attachment on plastic and laminin was significantly weaker. In summary, Lamc2-/- mouse recapitulates human JEB and provides novel insight into the role of laminin 5 in keratinocyte biology.
authors: Department of Dermatology and Cutaneous Biology, Jefferson Medical College, and Jefferson Institute of Molecular Medicine, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
authors: Xianmin
authors: X
authors: Meng
authors: John F
authors: JF
authors: Klement
authors: Dominic A
authors: DA
authors: Leperi
authors: David E
authors: DE
authors: Birk
authors: Takako
authors: T
authors: Sasaki
authors: Rupert
authors: R
authors: Timpl
authors: Jouni
authors: J
authors: Uitto
authors: Leena
authors: L
authors: Pulkkinen
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology; D001485:Basement Membrane / Q000502:physiology / Q000648:ultrastructure; D001768:Blister / Q000235:genetics / Q000473:pathology / Q000503:physiopathology; D002448:Cell Adhesion / Q000502:physiology; D015815:Cell Adhesion Molecules / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D004820:Epidermolysis Bullosa / Q000235:genetics* / Q000473:pathology / Q000503:physiopathology*; D005260:Female; D015870:Gene Expression; D006801:Humans; D007365:Intercellular Junctions / Q000502:physiology* / Q000648:ultrastructure; D007797:Laminin / Q000235:genetics*; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008854:Microscopy, Electron; D011247:Pregnancy; D012326:RNA Splicing; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000097794:Kalinin
chemical_list: D015815:Cell Adhesion Molecules; D007797:Laminin; D000097794:Kalinin; C400567:LAMC2 protein, human; C498522:Lamc2 protein, mouse