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Mitochondria frozen with trehalose retain a number of biological functions and preserve outer membrane integrity

R Yamaguchi1, A Andreyev, A N Murphy

  • 1Department of Molecular Immunology, La Jolla Institute for Allergy and Immunology, La Jolla, CA 92037, USA.

Cell Death and Differentiation
|September 16, 2006
PubMed

Insights

Trehalose preserves mitochondrial outer membrane integrity after freezing, enabling apoptosis research. Frozen mitochondria in trehalose buffer retain crucial functions, unlike those in standard buffers.

Area of Science:

  • Mitochondrial biology
  • Cell death research
  • Biochemistry

Background:

  • Bcl-2-family proteins control apoptosis by regulating mitochondrial outer membrane (MOM) integrity.
  • Studying MOM integrity in vitro requires functional mitochondria, typically freshly isolated.
  • Standard freezing methods compromise mitochondrial integrity, hindering apoptosis research.

Purpose of the Study:

  • To evaluate trehalose as a cryoprotectant for preserving mitochondrial function, particularly MOM integrity.
  • To determine if trehalose-frozen mitochondria remain responsive to Bcl-2-family proteins for apoptosis studies.

Main Methods:

  • Isolation of mouse liver mitochondria.
  • Freezing mitochondria in buffers with and without trehalose.
  • Assessment of MOM integrity, ultrastructure, and various mitochondrial functions (ATP synthesis, calcium swelling, membrane potential, protein import).

Main Results:

  • Mitochondria frozen in trehalose buffer maintained MOM integrity and responsiveness to Bcl-2-family proteins.
  • Trehalose preserved mitochondrial ultrastructure and functions like ATP synthesis, calcium-induced swelling, and protein import.
  • Minor reduction in bioenergetic function was observed in trehalose-frozen mitochondria.

Conclusions:

  • Trehalose effectively preserves mitochondrial MOM integrity and key biological functions after freezing.
  • Trehalose-frozen mitochondria are suitable for apoptosis research and other studies requiring intact MOM.
  • This method offers a valuable alternative for storing mitochondria for specific research applications.

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