Localization of laminin-5 in the epidermal basement membrane

被引:66
作者
Masunaga, T
Shimizu, H
Ishiko, A
Tomita, Y
Aberdam, D
Ortonne, JP
Nishikawa, T
机构
[1] KOSE CORP,BASIC RES LAB,TOKYO,JAPAN
[2] NIPPON KOKAN LTD CO HOSP,DEPT DERMATOL,KAWASAKI,KANAGAWA,JAPAN
[3] AKITA UNIV,SCH MED,DEPT DERMATOL,AKITA 010,JAPAN
[4] FAC MED NICE,INSERM U385,NICE,FRANCE
关键词
kalinin; nicein; GB3; epidermolysis bullosa; basement membrane; melanocyte; cryoultramicrotomy; immunoelectron microscopy; cryofixation; cryosubstitution;
D O I
10.1177/44.11.8918896
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Laminin-5 (kalinin/nicein/BM600) is a component of the epidermal basement membrane zone. Previous en bloc preembedding immunogold electron microscopy (EM) has shown that the epitope of GB3, a monoclonal antibody against laminin-5, is present in the lamina lucida (LL). However, precise localization of the entire laminin-5 molecule was unclear because of uneven and limited penetration of gold-labeled antibody in pre-embedding immunolabeling. In addition, the location of the GB3 epitope may not directly represent the location of the laminin-5 molecule itself. To elucidate the precise ultrastructural distribution of the entire laminin-5 molecule, we used polyclonal antibodies against different sites of laminin-5. Dual staining immunofluorescence with anti-laminin-5 and anti-melanocyte antibodies and immunoperoxidase EM showed that laminin-5 was present only beneath keratinocytes and not beneath melanocytes. Both cryoultramicrotomy and postembedding immunogold EM demonstrated that laminin-5 was localized to the lamina densa (LD) and the lower LL, with major labeling beneath the hemidesmosome. Quantitative analysis showed that 67-69% of gold particles were distributed to the LD and 88-90% were distributed beneath the hemidesmosome. Our results indicate that laminin-5 is localized mainly to the LD and partially to the lower LL, and is associated predominantly with hemidesmosomes.
引用
收藏
页码:1223 / 1230
页数:8
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