From the Second Department of Internal Medicine, Gifu University School
of Medicine (M.O., G.T., A.O., J.M., Y.H., S.M., H.F.), Gifu, Japan, and the
Department of Food Science, Kyoto Women's University (T.F.), Kyoto,
Japan.
Correspondence to Hisayoshi Fujiwara, MD, PhD, Second Department of Medicine, Gifu University School of Medicine, 40 Tsukasa-machi, Gifu 500, Japan.
Background—Modes of cell death have
been defined morphologically as apoptosis and oncosis.
Infarcted myocytes have been reported to show apoptosis, as
revealed by DNA fragmentation by DNA ladder and by in situ terminal
deoxynucleotidyl transferase–mediated dUTP nick
end-labeling (TUNEL) at the light microscopic level. We investigated
whether TUNEL-positive infarcted myocytes have apoptotic or
oncotic ultrastructures by using electron microscopic TUNEL, which can
simultaneously observe the ultrastructure and DNA
fragmentation of the same myocytes.
Methods and Results—Thirty rabbits were divided into 5 groups
(n=6 each) that were subjected to a sham operation or to 30-minute
ischemia followed by 0-minute, 30-minute, 2-hour, or 4-hour
reperfusion of a coronary artery. In the 2- and 4-hour
reperfusion groups only, DNA electrophoresis showed a ladder pattern,
and the light microscopic TUNEL finding was positive in the nuclei of
myocytes localized in the infarcted area (6±2% and 11±3%,
respectively). Electron microscopic TUNEL showed that nuclei with a
significant accumulation of immunogold particles (indicating an
electronic microscopic TUNEL–positive result) were observed only in
the infarcted myocytes with irreversibly oncotic ultrastructures that
were found in the hearts of the 2- and 4-hour reperfusion groups
(41±3% and 83±4%, respectively). Irreversibly oncotic myocytes
(indicated by swelling, inhomogeneously clumped chromatin
in nuclei, dense bodies in mitochondria, and/or ruptured plasma
membranes) were also seen in the 0- and 30-minute reperfusion groups,
which did not exhibit TUNEL-positive myocytes. There was no evidence of
apoptotic ultrastructures in the myocytes.
Conclusions—DNA fragmentation occurs in the myocytes that had
already shown irreversibly oncotic, but not apoptotic,
ultrastructures with ischemia and/or reperfusion. Therefore,
DNA fragmentation itself does not always mean apoptosis, and
so-called apoptotic infarcted myocytes may belong to a category
of cell death other than apoptosis.
© 1998 American Heart Association, Inc.
Basic Science Reports
"Apoptotic" Myocytes in Infarct Area in Rabbit Hearts May Be Oncotic Myocytes With DNA Fragmentation
Analysis by Immunogold Electron Microscopy Combined With In Situ Nick End-Labeling
Key Words: myocardial infarction • apoptosis • myocytes • ischemia • reperfusion
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