Quantitative analysis of UVB-induced apoptosis in human epidermis

S. A. Hurwitz, Dan F. Spandau

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

The quantitative measurement of the induction of apoptosis in cells grown in vitro can be accomplished using a variety of proven methods. However, the quantitative assay of apoptosis within an intact tissue is very laborious and the results can be misleading. We have established a method to quantitatively analyze the induction of apoptosis in human epidermis following UVB irradiation. The assay is based on the activation of the apoptotically induced enzyme caspase 3, using a synthetic caspase 3 substrate. The activation of caspase 3 was shown to correlate with the induction of apoptosis in human keratinocytes cultures as a monolayer. We then demonstrated that the activation of caspase 3 could be measured from UVB-irradiated whole skin. The induction of apoptosis was confirmed by cellular morphology and terminal deoxynucleotidyl transferase-mediated deoxyuridine triophosphate nick end-labeling. Therefore, we concluded that the measurement of caspase 3 specific activity in UVB-irradiated human epidermis was an efficient, inexpensive, and accurate method of quantitate UVB-induced apoptosis in vivo.

Original languageEnglish (US)
Pages (from-to)185-191
Number of pages7
JournalExperimental Dermatology
Volume9
Issue number3
DOIs
StatePublished - Jun 1 2000

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Keywords

  • Assay
  • Caspase
  • Flourescence
  • Keratinocyte
  • UVB

ASJC Scopus subject areas

  • Dermatology

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