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Mammalian sperm DNA susceptibility to in situ denaturation associated with the presence of DNA strand breaks as measured by the terminal deoxynucleotidyl transferase assay.
Abstract: Sperm from four mammalian species were analyzed by the sperm chromatin structure assay (SCSA) and the terminal deoxynucleotidyl transferase assay (TdTA) using flow cytometry. The SCSA quantitates the susceptibility of sperm nuclear DNA to in situ acid denaturation, while the TdTA quantitates the presence of endogenous DNA strand breaks in sperm nuclear chromatin. Correlations were seen between the percentage of sperm cells showing susceptibility to in situ acid denaturation and the percentage of cells showing the presence of DNA strand breaks for humans (r = 0.56, P = 0.004), rams (r = 0.84, P < 0.001), bulls (r = 0.78, P < 0.001), and stallions (r = 0.65, P < 0.001). No significant differences were seen when using fresh or frozen samples for either assay. These results suggest that sperm cells that are more susceptible to in situ DNA denaturation may have a greater number of accessible endogenous DNA strand breaks. We hypothesize that the DNA strand breaks produced in the normal transition from a somatic cell histone complex to a protamine complex are not ligated properly, resulting in residual DNA strand breaks and altered chromatin structure. Alternatively, altered chromatin structure could lead to the accessibility of the endogenous DNA strand breaks.
Publication Date: 1995-01-01 PubMed ID: 7768756 The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- Non-P.H.S.
Summary
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This research article investigates the DNA integrity of mammalian sperm, specifically assessing how susceptible the sperm’s DNA is to breaking under specific circumstances, and the potential reasons for these strand breaks. The study used sperm from four species, including humans, and found a correlation between susceptibility to DNA damage and the presence of DNA strand breaks within the sperm.
Overview of Study and Methodology
- The research utilized sperm samples from four different mammalian species – humans, rams, bulls, and stallions. The choice of subjects enabled the researchers to ensure the findings were not confined to a single species and explored the common genetic characteristics of these species.
- Two tests were performed on each of the samples – the sperm chromatin structure assay (SCSA) and the terminal deoxynucleotidyl transferase assay (TdTA). These assays determined the degree of susceptibility of the sperm to DNA damage (acid denaturation) and the presence of DNA strand breaks, respectively.
- Both these tests were conducted using flow cytometry, a technique used to measure and analyze the physical and chemical characteristics of particles in a fluid as they pass through at least one laser.
Primary Findings
- Upon conducting the assays, the results showed a correlation between the percentage of sperm cells susceptible to DNA damage and the percentage of cells showing the presence of DNA strand breaks. This statistical correlation exposes a relationship between the two factors within all species tested, indicating that the sperm cells which show increased susceptibility to DNA denaturation often have a higher number of strand breaks.
- No significant differences were identified between fresh and frozen samples. This implies the conditions and storage method of the sperm cells did not have a noticeable impact on the DNA susceptibility.
Potential Implications
- Beyond the observable trends and correlations, the authors proposed a hypothesis regarding the cause of the DNA strand breaks. They postulate that the normal transition from a somatic cell histone complex to a protamine complex, a process fundamental to sperm production, could cause unligated or improperly ligated DNA strand breaks, resulting in a subsequent alteration of the chromatin structure.
- Alternatively, they suggest that it is possible that the altered chromatin structure could lead to the strand breaks, rather than being a result of them. This underlines the complexity of the process and the challenges in defining clear cause-and-effect relationships.
This ground-breaking research adds to understanding of mammalian sperm DNA stability and the effects of DNA strand breaks on the health qualities of the sperm. These findings and hypotheses could be built upon to explore potential impacts on fertility, genetic integrity, and ultimately species propagation.
Cite This Article
APA
Sailer BL, Jost LK, Evenson DP.
(1995).
Mammalian sperm DNA susceptibility to in situ denaturation associated with the presence of DNA strand breaks as measured by the terminal deoxynucleotidyl transferase assay.
J Androl, 16(1), 80-87.
Publication
Researcher Affiliations
- South Dakota State University, Olson Biochemistry Laboratories, Brookings, USA.
MeSH Terms
- Animals
- Cattle
- Chromatin / metabolism
- DNA / metabolism
- DNA Damage
- DNA Nucleotidylexotransferase / metabolism
- Flow Cytometry
- Horses
- Humans
- Male
- Nucleic Acid Denaturation
- Sheep
- Spermatozoa / metabolism
Citations
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