A two-tailed Comet assay for assessing DNA damage in spermatozoa.
نویسندگان
چکیده
DNA fragmentation is considered an important parameter of semen quality, and of significant value as a predictor of male fertility. Poor quality chromatin is closely associated with, and highly indicative of, some fertility problems. Many methodologies to assess DNA fragmentation in spermatozoa are available, but they are all unable to differentiate between single-stranded DNA breaks (SSB) and double-stranded DNA breaks (DSB) in the same sperm cell. The two-tailed Comet assay (2T-Comet) protocol overcomes this limitation. A modification of the original Comet assay was developed for the simultaneous evaluation of DNA SSB and DSB in human spermatozoa. The 2T-Comet assay is a fast, sensitive, and reliable procedure for the quantification and characterization of DNA damage in spermatozoa. It is an innovative method for assessing sperm DNA integrity, which has important implications for human fertility and andrological pathology.
منابع مشابه
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متن کاملShortening of alkaline DNA unwinding time does not interfere with detecting DNA damage to mouse and human spermatozoa in the comet assay.
The comet assay was performed on mouse and human spermatozoa to examine the effect of alkaline DNA unwinding time. The spermatozoa were treated in vitro with the DNA-damaging agents, methyl methanesulfonate (MMS) or hydrogen peroxide (H₂O₂), and then embedded in agarose gel on glass slides. The slides were immersed in alkaline solution (> pH 13) for 1, 5, 10 and 20 min, and then subjected to th...
متن کاملShortening of alkaline DNA unwinding time does not interfere with detecting DNA damage to mouse and human spermatozoa in the comet
22 23 The Comet assay was performed on mouse and human spermatozoa to 24 examine the effect of alkaline DNA unwinding time. The spermatozoa were 25 treated in vitro with the DNA-damaging agents, methyl methanesulfonate 26 (MMS) or hydrogen peroxide (H2O2), and then embedded in agarose gel on 27 glass slides. The slides were immersed in alkaline solution (> pH 13) for 1, 5, 28 10, and 20 min, an...
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Background: Lymphocytes of Fanconi anemia (FA) show an increased sensitivity to the alkylating agents such as mitomycin C (MMC), but their responses to gamma-irradiation is controversial. The extent of DNA damage in leukocytes of FA patients following irradiation and MMC treatment was studied at cellular and single chromosome level. Methods: DNA damage induced by gamma-rays and MMC was measure...
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ورودعنوان ژورنال:
- Reproductive biomedicine online
دوره 18 5 شماره
صفحات -
تاریخ انتشار 2009