[Comparison between two methods for staining DNA of apoptotic spermatozoa].
Zhonghua Nan Ke Xue, 2005/2;11(2):101-3.
Yu WH[1], Li ZH, Yang WM, Li GZ, Zhou XM, Li KS
Affiliations
PMID: 15755026
Abstract
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.
conclusion: 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.
MeSH terms
Apoptosis; Benzimidazoles; Camptothecin; Cell Survival; DNA; Dose-Response Relationship, Drug; Enzyme Inhibitors; Fluorescent Dyes; Genistein; Humans; Male; Propidium; Sensitivity and Specificity; Spermatozoa; Staining and Labeling
More resources
EndNote: Download