Induction and inhibition of testicular germ cell apoptosis by fluoroacetate in rats.
Fluoroacetate (FA), an inhibitor of aconitase, is known to lower the intracellular level of adenosine triphosphate (ATP), which recently has been suggested to be a possible determinant of the form of cell death, apoptosis or necrosis. To investigate which form of germ cell death occurs in FA-induced testicular toxicity, adult Sprague Dawley rats were given a single oral dose of FA (0.5 or 1.0 mg/kg) and euthanized at 3, 6, 12, 24, 48, and 72 h thereafter. Germ cell degeneration was histologically first found in early round spermatids at stage I and in spermatogonia at stages II-IV of seminiferous tubules 6 and 12 h, respectively, after dosing. Degenerating spermatogonia exhibited characteristic features of apoptosis as demonstrated by both electron microscopy and in situ terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL), whereas spermatids did not. At the 24 and 48 h time points, degenerating spermatids were continually present and subsequently formed multinucleated giant cells, while the number of degenerating spermatogonia and TUNEL-labeled spermatogonia was drastically and/or significantly decreased compared to those from the control group, indicating that spontaneous male germ cell apoptosis is inhibited. Coincident with these morphological changes, DNA laddering on gel electrophoresis was apparent only 12 h after dosing. The results demonstrate that FA induces either apoptosis or necrosis of male germ cells in the early stage after dosing and subsequently inhibits spontaneous apoptosis.
Shinoda
K
K
Pathology Unit, Hita Laboratories, Chemicals Evaluation and Research Institute, Hita-shi, Oita, Japan. shinoda-kazutoshi@hita.cerij.or.jp
Mitsumori
K
K
Uneyama
C
C
Uehara
M
M
D000154:Aconitate Hydratase / Q000037:antagonists & inhibitors*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004734:Energy Metabolism / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005463:Fluoroacetates / Q000494:pharmacology*; D005854:Germ Cells / Q000187:drug effects* / Q000648:ultrastructure; D020287:In Situ Nick-End Labeling; D008297:Male; D008854:Microscopy, Electron; D009336:Necrosis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012671:Seminiferous Tubules / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D013087:Spermatids / Q000187:drug effects / Q000648:ultrastructure; D013737:Testis / Q000166:cytology* / Q000187:drug effects / Q000473:pathology
D004791:Enzyme Inhibitors; D005463:Fluoroacetates; D004247:DNA; D000154:Aconitate Hydratase