Comparison between two methods for staining DNA of apoptotic spermatozoa.
- Author:
Wen-Hui YU
1
;
Zhuo-Hua LI
;
Wei-Min YANG
;
Guang-Zhao LI
;
Xiao-Mei ZHOU
;
Kang-Sheng LI
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; Benzimidazoles; Camptothecin; pharmacology; Cell Survival; drug effects; DNA; metabolism; Dose-Response Relationship, Drug; Enzyme Inhibitors; pharmacology; Fluorescent Dyes; Genistein; pharmacology; Humans; Male; Propidium; Sensitivity and Specificity; Spermatozoa; drug effects; physiology; Staining and Labeling; methods
- From: National Journal of Andrology 2005;11(2):101-103
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo compare two fluorochrome staining methods for the assessment of sperm quality.
METHODSWashed 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.
RESULTSThe 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.
CONCLUSIONThe 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.
