Immunoelectron microscopic analysis of basement membrane components in type VII collagen knockout skin

VII型胶原基因敲除皮肤基底膜成分的免疫电镜分析

基本信息

批准号：
12670836
负责人：
ISHIKO Akira
金额：
$ 1.98万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670836/
关键词：
type VII collagen epidermolysis bullosa immunoelectron microscopy BPAG2 laminin 5 basement membrane mutation analysis

项目摘要

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.

营养不良性大疱性表皮松解症 (RDEB) 是一种先天性大疱性疾病，由缺乏将表皮基底膜（致密层）固定到真皮的 VII 型胶原蛋白引起。 VII 型胶原蛋白与其他表皮基底膜分子之间的分子关系尚未完全阐明。本研究的目的是利用 RDEB 患者皮肤（即 VII 型基因敲除人类皮肤）阐明 VII 型胶原蛋白与其他基底膜分子之间的体内分子超微结构关系。为此，我们比较了 VII 型胶原蛋白敲除人体皮肤和正常人体皮肤之间基底膜分子（包括 VII 型胶原蛋白、BPAG2、层粘连蛋白 5）的精确超微结构定位。 RDEB 的诊断通过电子显微镜证实，并且缺乏 LH7.2 抗原、VII 型胶原蛋白 N 末端结构域的表达。对五名 RDEB 患者的血液样本进行基因组 DNA 洗脱处理。扩增编码 VII 型胶原蛋白的 COL7A1 基因并分析 DNA 序列。在 10 个等位基因中，鉴定出 4 个等位基因的 3 个突变：6573+1G>C、5504delA 和 5818delC。经同意，从三名 RDEB 患者身上获取皮肤样本。用包埋后免疫金电子显微镜观察BPAG2和层粘连蛋白5的N和C末端定位，并与正常人皮肤进行统计比较。结果，BPAG2 的 N 末端位于半桥粒，层粘连蛋白 5 位于致密层。正常皮肤和 RDEB 皮肤之间这些分子的定位没有显着差异。然而，BPAG2 的 C 末端显着从致密层转移到表皮侧。这些结果表明，VII 型胶原蛋白可能通过 BPAG2 的 C 端结构域与 BPAG2 之间存在一些分子相互作用，这两者都位于正常皮肤的致密层。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Akira Ishiko, Hiroshi Shimizu: "Electron microscopy in diagnosis of autoimmune bullous disorders"Clinics in Dermatology. 19. 631-637 (2001)

Akira Ishiko、Hiroshi Shimizu：“电子显微镜诊断自身免疫性大疱性疾病”皮肤科诊所。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Akira Ishiko and Hiroshi Shimizu: "Electron microscopy in diagnosis of autoimmune bullous disorders"Clinics in Dermatology. 19. 631-637 (2001)

Akira Ishiko 和 Hiroshi Shimizu：“电子显微镜诊断自身免疫性大疱性疾病”皮肤科诊所。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Yasuko Takizawa, Yoshiki Hiraoka, Hayato Takahashi, Akira Ishiko, Isamu Yasuraoka, Isao Hashimoto, Sadakazu Also Takeji Nishikawa, Hiroshi Shimizu: "Compound heterozygosity for a point mutation and a deletion located at slice acceptor sited I the LAMB3 ge

Yasuko Takizawa、Yoshiki Hiraoka、Hayato Takahashi、Akira Ishiko、Isamu Yasuraoka、Isao Hashimoto、Sadakazu Also Takeji Nishikawa、Hiroshi Shimizu：“位于 LAMB3 ge 切片受体位点的点突变和缺失的复合杂合性