2001 Fiscal Year Final Research Report Summary
Immunoelectron microscopic analysis of basement membrane components in type VII collagen knockout skin
| Project/Area Number |
12670836
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Dermatology
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| Research Institution | Keio University |
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Principal Investigator |
ISHIKO Akira Keio University School of Medicine, Dermatology, Assistant professor, 医学部, 専任講師 (10202988)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Hiroshi Hokkaido University School of Medicine, Dermatology, Professor, 医学部, 教授 (00146672)
SUZUKI Yosuke Keio University School of Medicine, Dermatology, Researcher, 医学部, 助手 (90306845)
NISHIKAWA Takeji Keio University School of Medicine, Dermatology,, Professor, 医学部, 教授 (50051579)
MASUNAGA Takuji KOSE corporation Fundamental Research Laboratory, Chief researcher, 研究本部・基礎研究所, 研究員
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| Project Period (FY) |
2000 – 2001
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| Keywords | type VII collagen / epidermolysis bullosa / immunoelectron microscopy / BPAG2 / laminin 5 / basement membrane / mutation analysis |
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| Research Abstract |
Receive dystrophic epidermolysis bullosa (RDEB) is a congenital bullous disease caused by lack of type VII collagen that anchors epidermal basement membrane (lamina densa) to the dermis. Molecular relationship between the type VII collagen and the other epidermal basement membrane molecules is not fully elucidated. The purpose of this study is to elucidate the in vivo molecular ultrastructural relationship between the type VII collagen and the other basement membrane molecules using the skin of RDEB patients, which were the type VII knockout human skin. For this, the precise ultrastructural localization of basement membrane molecules including type VII collagen, BPAG2, laminin 5 were compared between the type VII collagen knockout human skin and the normal human skin. The diagnosis of RDEB confirmed by electron microscopy and the lack of expression of LH7.2 antigen, N-terminus domain of type VII collagen. Blood samples from the five patients with RDEB were processed for genomic DNA elution. COL7A1 gene that encodes type VII collagen was amplified and DNA sequences were analyzed. Among ten alleles, three mutations on four alleles were identified: 6573+1G>C, 5504delA and 5818delC. Skin samples were obtained from three RDEB patients by consent. Localizations of N and C terminus of BPAG2, and laminin 5 were observed with post-embedding immunogold electron microscopy and compared statistically with those of normal human skin. As results, the N terminus of BPAG2 was localized at the hemidesmosome and laminin 5 was localized at lamina densa. There was no significant difference in localization of these molecules between normal and RDEB skin. However, C-terminus of BPAG2 were significantly shifted from lamina densa to the epidermal side. These results indicated that type VII collagen may have some molecular interaction between BPAG2 via C-terminal domain of BPAG2, both of which were localized at lamina densa in the normal skin.
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