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Journal of Histochemistry and Cytochemistry, Vol. 49, 1277-1284, October 2001, Copyright © 2001, The Histochemical Society, Inc.

ARTICLE

Mitochondrial Transmembrane Potential Changes Support the Concept of Mitochondrial Heterogeneity During Apoptosis

Dmitri V. Kryskoa, Frank Roelsa, Luc Leybaertb, and Katharina D'Herdea
a Department of Human Anatomy, Embryology, Histology and Medical Physics, Ghent University, Ghent, Belgium
b Department of Physiology and Pathophysiology, Ghent University, Ghent, Belgium

Correspondence to: Dmitri V. Krysko, Dept. of Human Anatomy, Embryology, Histology and Medical Physics, Faculty of Medicine, Ghent University, Godshuizenlaan 4, B-9000 Ghent, Belgium. E-mail: Dmitri_Krysko@yahoo.com

Dissipation of mitochondrial membrane potential ({Delta}{Psi}m) and release of cytochrome c from mitochondria appear to be key events during apoptosis. The precise relationship (cause or consequence) between both is currently unclear. We previously showed in a model of serum-free cultured granulosa explants that cytochrome c is retained in a subset of respiring mitochondria until late in the apoptotic process. In this study we further investigated the issue of heterogeneity by using the {Delta}{Psi}m-sensitive probe CM-H2TMRos in combination with a DNA fluorochrome. Changes of {Delta}{Psi}m were assessed qualitatively by epifluorescence microscopy and were quantified using digital imaging microscopy. This approach yielded the following results: (a) CM-H2TMRos staining is a reliable and specific procedure to detect {Delta}{Psi}m changes in granulosa cells explants; (b) dissipation of transmembrane potential is an early event during apoptosis preceding nuclear changes but is confined to a subpopulation of mitochondria within an individual cell; (c) in frankly apoptotic cells a few polarized mitochondria can be detected. These findings support the hypothesis that ATP needed for completion of the apoptotic cascade can be generated during apoptosis in a subset of respiring mitochondria and is not necessarily derived from anaerobic glycolysis. (J Histochem Cytochem 49:1277–1284, 2001)

Key Words: mitochondrial membrane, potential, apoptosis, granulosa cells, CM-H2TMRos, cytochrome c


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