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Related Experiment Videos

Multiple proteases are involved in thymocyte apoptosis

B Zhivotovsky1, A Gahm, M Ankarcrona

  • 1Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden.

Experimental Cell Research
|December 1, 1995
PubMed
Summary

Protease inhibitors targeting interleukin-1 beta-converting enzyme (ICE) prevent apoptosis in rat thymocytes induced by methylprednisolone or etoposide. A separate protease inhibitor blocked apoptosis only with methylprednisolone treatment.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Apoptosis, or programmed cell death, is a crucial biological process.
  • Proteases, enzymes that break down proteins, are implicated in regulating apoptosis.
  • Specific proteases, like ICE and Ca(2+)-regulated serine proteases, are investigated for their roles.

Purpose of the Study:

  • To elucidate the specific roles of interleukin-1 beta-converting enzyme (ICE)-like proteases and Ca(2+)-regulated serine proteases in methylprednisolone- and etoposide-induced thymocyte apoptosis.
  • To determine the temporal involvement of these proteases during the apoptotic cascade.

Main Methods:

  • Rat thymocytes were treated with methylprednisolone or etoposide.
  • Selective substrate inhibitors, VADcmk (for ICE-like proteases) and AAPFcmk (for Ca(2+)-regulated serine proteases), were used.

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  • Apoptotic markers including lamin proteolysis, chromatin fragmentation, and morphological changes were assessed.

    • Main Results:

    • VADcmk significantly inhibited apoptosis markers in thymocytes treated with both methylprednisolone and etoposide.
    • AAPFcmk inhibited apoptosis markers in methylprednisolone-treated thymocytes but not in etoposide-treated ones.
    • ICE-like protease activity was elevated early in apoptosis induced by both agents and was inhibited by VADcmk.

    Conclusions:

    • ICE-like protease activity is essential for the early stages of apoptosis induced by both methylprednisolone and etoposide in thymocytes.
    • Ca(2+)-regulated serine protease is a necessary component of the apoptotic pathway specifically for methylprednisolone-induced apoptosis.