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Related Experiment Videos

Nuclear apoptosis induced by isolated mitochondria.

Z F Jiang1, Y Zhao, X Hong

  • 1College of Life Sciences, Peking University, Beijing, China.

Cell Research
|October 14, 2000
PubMed
Summary

Mitochondria can trigger apoptosis in isolated nuclei. Mitochondrial supernatants, even without cytosolic factors, induce nuclear DNA fragmentation and chromatin condensation, mediated by a novel endonuclease.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Mitochondria play a crucial role in cellular processes, including apoptosis.
  • The precise mechanisms by which mitochondria initiate apoptotic events in isolated nuclei are not fully understood.

Purpose of the Study:

  • To investigate the ability of isolated mitochondria and their components to induce apoptosis in isolated nuclei.
  • To characterize the factors responsible for mitochondrial-induced nuclear apoptosis.

Main Methods:

  • Isolation and purification of mitochondria from mouse liver and spinach leaves.
  • Incubation of isolated mitochondria with Xenopus laevis egg extracts and isolated mouse liver nuclei.
  • Preparation of mitochondrial supernatants through hypotonic treatment and high-speed centrifugation.

Related Experiment Videos

  • Assessment of apoptotic changes in nuclei, including chromatin condensation, margination, and DNA laddering.
  • Inhibition studies using caspase inhibitors (AC-DEVD-CHO, AC-YVAD-CHO).
  • Characterization of the nuclear endonuclease activity.

    • Main Results:

    • Isolated mitochondria induced apoptotic changes in mouse liver nuclei within Xenopus egg extracts.
    • Mitochondrial supernatants alone, without cytosolic factors, triggered nuclear apoptosis.
    • Apoptotic changes included chromatin condensation, margination, and DNA fragmentation.
    • Caspase inhibitors partially blocked DNA fragmentation and fully blocked chromatin condensation.
    • A novel nuclear endonuclease, distinct from known enzymes like endonuclease G, was identified in the mitochondrial supernatants.

    Conclusions:

    • Mitochondria contain factors capable of directly inducing apoptosis in isolated nuclei.
    • A novel endonuclease, activated or released by mitochondria, plays a key role in DNA fragmentation during this process.
    • These findings shed light on the intrinsic pathway of apoptosis initiation by mitochondria.