PMID: 11915365Mar 28, 2002Paper

Evaluation of mitochondrial function and membrane integrity by dual fluorescent staining for assessment of sperm status in rats

The Journal of Toxicological Sciences
Masashi KatoYoichi Nagamura

Abstract

Dual fluorescent staining (DFS) with calcein acetoxy methyl ester (CAM), which labels the cellular esterase activity that is a major component of energy metabolism in cellular mitochondria, and with ethidium homodimer-1 (EthD-1) was used to evaluate mitochondrial function and membrane integrity in rat spermatozoa. The spermatozoa stained by DFS could be classified into three different populations microscopically when excited at 490 nm after 60 min incubation. 1) Spermatozoa, which were stained with CAM alone and had maintained either mitochondrial function or membrane integrity, were identified as live during incubation. 2) Spermatozoa, which were stained with EthD-1 alone and had lost either mitochondrial function or membrane integrity, were identified as already dead at the beginning of incubation. 3) Spermatozoa, which were stained with both CAM and EthD-1 and had maintained mitochondrial function with membrane breached, were identified as having died during incubation. Two toxicological tests, an in vitro triton X-100 experiment and an in vivo nitrobenzene experiment, were done. All spermatozoa were immobilized and lost either mitochondrial function or membrane integrity by 1.0% triton X-100 treatment. Almost no motile sper...Continue Reading

References

Aug 1, 1995·Biology of Reproduction·D L Garner, L A Johnson
Apr 16, 1998·Reproductive Toxicology·C M VetterT R Skopek
Jan 29, 1999·The Journal of Toxicological Sciences·T YamamotoM Takeuchi
Mar 21, 2001·The Journal of Toxicological Sciences·Y BanY Ooshima

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