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Volume 17, Number 11,
Issue of June 1, 1997
pp. 3981-3989
Copyright ©1997 Society for Neuroscience
The In Vivo Time Course for Elimination of
Adrenalectomy-Induced Apoptotic Profiles from the Granule Cell Layer of
the Rat Hippocampus
Received Dec. 19, 1996; revised Feb. 18, 1997; accepted March 10, 1997.
Zhongting Hu,
Kazunari Yuri,
Hitoshi Ozawa,
Haiping Lu, and
Mitsuhiro Kawata
Department of Anatomy and Neurobiology, Kyoto Prefectural
University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602, Japan
Although apoptotic cellular degeneration has been reported to be
extremely rapid with the use of in vitro models, the
time needed to clear apoptotic neurons in the in vivo
brain is unknown. In this study we used a simple morphological approach
to solve this problem. Four days after adrenalectomy (ADX), all of the operated rats morphologically displayed hippocampal granule cell apoptosis that was prevented completely by corticosterone replacement immediately after ADX. Therefore, we intravenously injected the rats
with corticosterone 4 d after ADX and subsequently maintained them
on corticosterone replacement in saline drinking water. This corticosterone replacement could protect healthy granule cells promptly
and continuously against hormone-deficient apoptosis, because the
normal glucocorticoid receptor immunoreactivity within the granule cell
nuclei, which disappeared after ADX, was identified 1 hr after
corticosterone replacement was started, and this effect persisted for
several days. However, this corticosterone treatment could not prevent
the irreversible apoptosis of the already degenerated granule cells at
various stages of the same progressive apoptotic process. Then we
successively traced the disappearance of apoptotic granule cells
throughout the hippocampus at different time points by Nissl and silver
staining. Given that the apoptotic cells at the earliest stage of the
degenerating process when the ADX rats received corticosterone
injection were the last to disappear, the period from corticosterone
injection until the disappearance of the last degenerating debris of
apoptotic cells was taken to represent the time course for elimination
of apoptotic neurons in vivo. We discovered that the
elimination of apoptotic granule cells took 72 hr.
Key words:
adrenalectomy;
corticosterone;
granule cell;
apoptosis;
silver and Nissl staining;
chromatin condensation;
time course
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