Document Detail

Immunohistochemistry and in situ hybridization in the study of human skin melanocytes.
MedLine Citation:
PMID:  17286807     Owner:  NLM     Status:  MEDLINE    
Although keratinocytes are the most numerous type of cell in the skin, melanocytes are also key players as they produce and distribute melanin that protects the skin from ultraviolet (UV) radiation. In vitro experiments on melanocytic cell lines are useful to study melanogenesis and their progression towards melanoma. However, interactions of melanocytes with keratinocytes and with other types of cells in the skin, such as fibroblasts and Langerhans cells, are also crucial. We describe two techniques, immunohistochemistry (IHC) and tissue in situ hybridization (TISH), that can be used to identify and study melanocytes in the skin and their responses to UV or other stimuli in situ. We describe a practical method to localize melanocytic antigens on formalin-fixed, paraffin-embedded tissue sections and in frozen sections using indirect immunofluorescence with conjugated secondary antibodies. In addition, we detail the use of TISH and its combination with IHC to study mRNA levels of genes expressed in the skin at cellular resolution. This methodology, along with relevant tips and troubleshooting items, are important tools to identify and study melanocytes in the skin.
Thierry Passeron; Sergio G Coelho; Yoshinori Miyamura; Kaoruko Takahashi; Vincent J Hearing
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Intramural; Review    
Journal Detail:
Title:  Experimental dermatology     Volume:  16     ISSN:  0906-6705     ISO Abbreviation:  Exp. Dermatol.     Publication Date:  2007 Mar 
Date Detail:
Created Date:  2007-02-08     Completed Date:  2007-03-15     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9301549     Medline TA:  Exp Dermatol     Country:  Denmark    
Other Details:
Languages:  eng     Pagination:  162-70     Citation Subset:  IM    
Pigment Cell Biology Section, Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
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MeSH Terms
In Situ Hybridization*
Melanocytes / chemistry*,  cytology*
Skin / cytology*

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