Document Detail


Phorbol ester- and calcium-induced reorganization of 180-kDa bullous pemphigoid antigen on the ventral surface of cultured human keratinocytes as studied by immunofluorescence and immunoelectron microscopy.
MedLine Citation:
PMID:  1426040     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The hemidesmosome is an adhesion structure of the epidermal-dermal junction in keratinocytes. When keratinocytes migrate laterally or upward to differentiate, they must control the formation and disintegration of the hemidesmosomes. When keratinocytes are cultured in low-calcium (below 0.1 mM) medium, all cells behave like basal cells, adhere to the culture dish, and proliferate without differentiation. The calcium addition induces the differentiation. A bullous pemphigoid antigen, 180-kDa BPA, has been shown to be a component of the hemidesmosome. Using a monoclonal antibody to the 180-kDa BPA and a human squamous cell carcinoma cell line (DJM-1 cells), the fate of hemidesmosomes was studied after the addition of calcium to low-calcium-grown cells and 12-tetradecanoylphorbol-13-acetate (TPA) to high-calcium (1.87 mM) grown cells by immunofluorescence and immunoelectron microscopy. The antigen was distributed evenly as fine dots on the entire ventral surface of low-calcium cells, whereas they formed a peculiar, concentric ring or arch arrangement on the ventral surface of high-calcium cells. Immunoelectron microscopy revealed the deposits of gold particles at sites on the membrane surface, where some filamentous or electron-dense materials were associated, although the complete structure of hemidesmosomes was not formed. They deposited directly onto the membrane surface in low-calcium cells and with a distance of 20-50 nm from the membrane surface in high-calcium cells. The calcium addition caused a profound reduction of the 180-kDa BPA-positive area for 30 to 120 min and then formed the high-calcium-ring pattern after 4 to 6 h. A similar calcium response was seen in normal human keratinocytes. TPA (16 nM) treatment caused disintegration of the ring pattern in high-calcium DJM-1 cells. This was inhibited with a protein kinase C (PKC) inhibitor. H7 (20 microM). These results suggest that the hemidesmosome is a dynamic structure and PKC can be one of the major factors in controlling the hemidesmosome, since it is known that the low-high calcium shift induces a calcium influx and a PKC activation, and TPA activates PKC in keratinocytes.
Authors:
Y Kitajima; K Owaribe; Y Nishizawa; Y Jokura; H Yaoita
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Experimental cell research     Volume:  203     ISSN:  0014-4827     ISO Abbreviation:  Exp. Cell Res.     Publication Date:  1992 Nov 
Date Detail:
Created Date:  1992-12-04     Completed Date:  1992-12-04     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0373226     Medline TA:  Exp Cell Res     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  17-24     Citation Subset:  IM    
Affiliation:
Department of Dermatology, Jichi Medical School, Tochigiken, Japan.
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MeSH Terms
Descriptor/Qualifier:
1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
Autoantigens / analysis,  metabolism*
Calcium / pharmacology*
Carcinoma, Squamous Cell
Carrier Proteins*
Cell Membrane / drug effects,  metabolism,  ultrastructure
Cells, Cultured
Collagen*
Cytoskeletal Proteins*
Desmosomes / drug effects,  metabolism*,  ultrastructure
Fluorescent Antibody Technique
Humans
Isoquinolines / pharmacology
Keratinocytes / drug effects,  metabolism*,  pathology
Microscopy, Immunoelectron
Molecular Weight
Nerve Tissue Proteins*
Non-Fibrillar Collagens*
Pemphigoid, Bullous / pathology*
Piperazines / pharmacology
Protein Kinase C / antagonists & inhibitors
Skin Neoplasms
Tetradecanoylphorbol Acetate / pharmacology*
Tumor Cells, Cultured
Chemical
Reg. No./Substance:
0/Autoantigens; 0/Carrier Proteins; 0/Cytoskeletal Proteins; 0/DST protein, human; 0/Isoquinolines; 0/Nerve Tissue Proteins; 0/Non-Fibrillar Collagens; 0/Piperazines; 0/collagen type XVII; 16561-29-8/Tetradecanoylphorbol Acetate; 7440-70-2/Calcium; 84477-87-2/1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; 9007-34-5/Collagen; EC 2.7.11.13/Protein Kinase C

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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