[Comparison between two methods for staining DNA of apoptotic spermatozoa].
OBJECTIVE
To compare two fluorochrome staining methods for the assessment of sperm quality.


METHODS
Washed sperm cells were incubated in 0, 0.15, or 15 micromol/L camptothecin (CAM), or 0.37 or 3.7 mmol/L genistein (GEN) at 37 degrees C for 4 hours. The sperm cells were analyzed for cycle-independent apoptosis and necrosis by single-stain compared with dual-stain fluorescence microscopy to contrast the relative effectiveness of these two approaches.


RESULTS
The single-stain procedure could not detect the sperm viability differences. In contrast, the dual-stain procedure identified a dosage-dependent decrease in the viability and increased necrozoospermia after topoisomerase inhibitor CAM and GEN treatments. Apoptosis was 2-fold higher with topoisomerase inhibitor treatment.


CONCLUSIONS
The two topoisomerase inhibitors were associated with increased apoptosis and dosage-dependent necrosis. The data suggested that the dual-stain combination Hoechst 33342/PI was more sensitive than the single Hoechst 33342 stain analysis and permitted quantitative analysis of the apoptosis and necrosis in sperm.
Yu
Wen-Hui
WH
Department of Laboratory, Shenzhen Hospital of Traditional Chinese Medicine, Shenzhen, Guangdong 518033, China. YWHSTGD@sina.com
Li
Zhuo-Hua
ZH
Yang
Wei-Min
WM
Li
Guang-Zhao
GZ
Zhou
Xiao-Mei
XM
Li
Kang-Sheng
KS
D017209:Apoptosis / Q000187:drug effects; D001562:Benzimidazoles; D002166:Camptothecin / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005456:Fluorescent Dyes; D019833:Genistein / Q000494:pharmacology; D006801:Humans; D008297:Male; D011419:Propidium; D012680:Sensitivity and Specificity; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*; D013194:Staining and Labeling / Q000379:methods*
D001562:Benzimidazoles; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D011419:Propidium; D004247:DNA; D019833:Genistein; C017807:bisbenzimide ethoxide trihydrochloride; D002166:Camptothecin