Cryopreservation of Ram Semen Facilitates Sperm DNA Damage: Relationship Between Sperm Andrological Parameters and the Sperm Chromatin Structure Assay

Analysis of sperm parameters is very important for predicting the outcome of assisted reproductive techniques and is necessary for determination of fertility potential of males tested for artificial insemination. In our study we have determined the level of bull and boar sperm DNA damage by Sperm Chromatin Structure Assay (SCSA). This test is based on increased susceptibility of altered DNA (strand breaks) in sperm nuclear chromatinto in situ denaturation measured by flow cytometry after staining with acridine orange (AO). Sperm chromatin damage was quantified by percentages of spermatozoa with detectable DNA Fragmentation Index – DFI divided into moderate (m-DFI) and high (h-DFI) DFI. Percentage of immature cells (HDS; cells with High DNA Stainability) was also evaluated. We measured sperm SCSA parameters in a total of 37 bulls in two groups from different localities and 68 boar samples from one locality. Significantly higher percentage of spermatozoa with detectable DFI was detected in six bulls (16.2%) and a significantly higher percentage of immature cell forms (HDS) was found in other six bulls (16.2%) among all tested bulls. The mean percentages of spermatozoa with h-DFI and HDS of bulls from the second group were statistically higher than those from the first group (P < 0.01). Five boars (7.4%) of all tested boars had significantly higher percentage of spermatozoa with DFI and 18 boars (26.5%) had significantly higher percentage of sperm with HDS compared to the other boars. Both percentages of spermatozoa with DFI and HDS were significantly higher in one boar compared to the others. Boars had significantly higher percentages of spermatozoa with h-DFI and HDS (P < 0.0001) in comparison to bulls. For individual bulls, the highest percentages of spermatozoa with DFI and HDS were 20.8% and 3.5%, respectively while for boars these were 17.6% and 10.2%, respectively. No significant correlations were found between percentages of spermatozoa with DFI and HDS. This sensitive procedure seems to be convenient as additional method for semen quality detection in farm animals before their exploitation in breeding.

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Evaluating human sperm DNA integrity: relationship between 8-hydroxydeoxyguanosine quantification and the sperm chromatin structure assay

Zygote ◽

10.1017/s0967199403002442 ◽

2003 ◽

Vol 11 (4) ◽

pp. 367-371 ◽

Cited By ~ 49

Author(s):

Isabelle Oger ◽

Christelle Da Cruz ◽

Gilles Panteix ◽

Yves Menezo

Keyword(s):

Dna Fragmentation ◽

Chromatin Structure ◽

Inverse Relationship ◽

Sperm Concentration ◽

Sperm Chromatin ◽

Sperm Chromatin Structure Assay ◽

Fragmentation Index ◽

Sperm Dna ◽

Oxidative Products ◽

Dna Fragmentation Index

In our work, we have used 8-hydroxy-deoxyguanosine (8-OH-dG), one of the major oxidative products of sperm DNA, in a population of patients consulting for infertility. We found an inverse relationship between sperm concentration and the log of the ratio of 8-OH-dG to dG (P<0.01). On the same patients' sperm samples, the sperm chromatin structure assay (SCSA) was performed. An inverse relationship was observed between the DNA fragmentation index and sperm concentration (P<0.001). There was also a positive relationship between SCSA and log 8-OH-dG/dG. This indicates that DNA fragmentation measured by the SCSA originates in part from oxidative products. In a few patients, antioxidant treatment decreased the DNA fragmentation index below the threshold of 30% that is crucial for subfertility.

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