Does conventional freezing affect sperm DNA fragmentation?

Lê, Minh Tâm; Nguyen, Thai Thanh Thi; Nguyen, Tung; Nguyen, Trung Van; Nguyen, Tam An Thi; Nguyen, Quoc Huy Vu; Cao, Thanh Ngoc

doi:10.5653/cerm.2019.46.2.67

Cited by 33 publications

(22 citation statements)

References 41 publications

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“…Despite the extensive progress that has been made in the field, decreases in sperm motility and DNA integrity are commonly observed after cryopreservation 30,39 . In this report we found that rapid freezing using R18S3 + MTG also significantly reduced sperm motility and increased DNA fragmentation in both wildtype and mutant mouse strains, observations consistent with previous results in mice 26,29 and humans [30][31][32]40 . In this report, we also found that DFI of frozen-thawed sperm from mutant mouse strains was consistently and highly correlated with male mouse fertility compared to traditional measures of sperm quality (count, morphology, and motility).…”

Section: Discussionsupporting

confidence: 92%

DNA fragmentation index (DFI) as a measure of sperm quality and fertility in mice

Lloyd

2020

Sci Rep

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Although thousands of genetically modified mouse strains have been cryopreserved by sperm freezing, the likelihood of cryorecovery success cannot be accurately predicted using conventional sperm parameters. The objective of the present study was to assess the extent to which measurement of a sperm DNA fragmentation index (DFI) can predict sperm quality and fertility after cryopreservation. Using a modified TUNEL assay, we measured and correlated the DFI of frozen-thawed sperm from 83 unique mutant mouse strains with sperm count, motility and morphology. We observed a linear inverse correlation between sperm DFI and sperm morphology and motility. Further, sperm DFI was significantly higher from males with low sperm counts compared to males with normal sperm counts (p < 0.0001). Additionally, we found that viable embryos derived using sperm from males with high DFI (62.7 ± 7.2% for IVF and 73.3 ± 8.1% for ICSI) failed to litter after embryo transfer compared to embryos from males with low DFI (20.4 ± 7.9% for IVF and 28.1 ± 10.7 for ICSI). This study reveals that measurement of DFI provides a simple, informative and reliable measure of sperm quality and can accurately predict male mouse fertility.

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Section: Discussionsupporting

confidence: 92%

DNA fragmentation index (DFI) as a measure of sperm quality and fertility in mice

Lloyd

2020

Sci Rep

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“…Decreased motility is also caused by damage to mitochondria that affects the production of adenosine triphosphate (ATP), which is the energy resource required for sperm to be motile. Decreased sperm motility is caused by mitochondrial damage [22]. Sperm mitochondria are composed of matrix helices located in the middle part (mid-piece).…”

Section: Discussion the Post-freeze Thawing Quality Of Holstein Bull Sperm And The Correlation With Fertility Levelmentioning

confidence: 99%

Sperm protein markers for Holstein bull fertility at National Artificial Insemination Centers in Indonesia

et al. 2020

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Background and Aim: Holstein cows and heifers are widely bred in Indonesia by artificial insemination (AI) to increase population and milk production. Sperm fertility is modulated by genetic factors, but the analysis of sperm quality is still based on macro- and microscopic characteristics. This study aimed to analyze both sperm quality and proteins of Holstein bulls at different fertility levels. Materials and Methods: The frozen semen samples were collected from the Indonesia National AI Center. They were classified based on the reproductive efficiency data and were grouped into high fertile (HF) and low fertile (LF). Sperm qualities were evaluated by microscopic evaluation. The Holstein sperm proteins were extracted using phenylmethanesulfonyl fluoride as a protease inhibitor and the benzidine detergent extraction method. Discontinuous sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was conducted to analyze the molecular weights (MWs) of the sperm proteins. The data obtained were analyzed by a t-test using the one-factor bull fertility level, and Spearman's correlation analysis was used to identify the correlation between the sperm microscopic evaluation parameters and protein bands. Results: The sperm motility post-freeze thawing was not significantly different between the HF and LF (p>0.05). The HF level had a higher percentage of viability, intact plasma membrane integrity, and intact acrosomes than the LF (p<0.05). Five protein bands were found in the SDS-PAGE of sperm proteins of Holstein bulls with different concentrations. Sperm proteins with MWs of 17.51 kDa, 14.87 kDa, 33.71 kDa, and 41.97 kDa were abundant in the Holstein bulls with an HF level, while 55 kDa proteins were abundant in the LF level of Holstein bulls. The sperm of Holstein bulls in the HF level contained proteins of about 33.71 kDa that were not detected in the LF. Conclusion: The sperm protein with a molecular weight of 33.71 kDa was predicted to be a specific protein biomarker that influences bull fertility. Sperm fertilization abilities were also determined by the sperm proteins, the morphology of sperm acrosomes, and the quality of plasma membranes. This method can be used to select bulls with high fertility to increase the population of Holstein bulls.

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“…One of the products of lipid peroxidation is MDA [ 6 ]. Free radicals of ROS may cause lipid peroxidation and mitochondrial and nuclear DNA base modifications [ 40 ]. Modification of 8-OH-guanine or 8-OH-20-deoxyguanosine will destabilize the DNA structure and causes SDF [ 8 ].…”

Section: Discussionmentioning

confidence: 99%

Green tea extract increases the quality and reduced DNA mutation of post-thawed Kacang buck sperm

Mustofa

Susilowati

Wurlina

et al. 2021

Heliyon

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The study aimed to determine the addition of green tea extract (GTE) in extender on the quality and DNA mutation of post-thawed Kacang buck sperm. The sperm DNA mutation was observed on nicotinamide adenine dinucleotide hydride (NADH) dehydrogenase 1 (ND1) of mitochondrial Deoxyribonucleic Acid (mtDNA). A pool of 12 Kacang buck ejaculates was diluted in skim milk-egg yolk extender contained 0, 0.05, 0.10, and 0.15 mg of GTE/100 mL for T0, T1, T2, and T3 group, respectively. Each of the aliquot groups was packaged in 0.25 mL French mini straw contained 60 million alive sperm and froze according to the protocol. The ND1 mtDNA amplification of samples was carried out Polymerase Chain Reaction machine, followed by DNA sequencing using the Sanger method. Meanwhile, the phylogenetic tree was constructed using the neighbor-joining (NJ) method with MEGA 7.0 software. The results showed that the T2 group maintained the highest quality for Kacang buck post-thawed semen. There was the highest percentages of sperms viability, motility, intact plasma membrane (IPM), the lowest of malondialdehyde (MDA) concentration, sperm DNA fragmentation (SDF), the total and types of ND1 mtDNA mutation frequency. The phylogenetic tree analysis revealed that the clade of the T2 group was most closely related to the sequence reference. However, there was no correlation between the semen quality parameters (sperm viability, motility, IPM, MDA concentration, and SDF) with ND1 mtDNA mutation of post-thawed Kacang buck semen. It could be concluded that GTE was useful as an antioxidant for Kacang buck semen extender for frozen sperm.

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Does conventional freezing affect sperm DNA fragmentation?

Cited by 33 publications

References 41 publications

DNA fragmentation index (DFI) as a measure of sperm quality and fertility in mice

DNA fragmentation index (DFI) as a measure of sperm quality and fertility in mice

Sperm protein markers for Holstein bull fertility at National Artificial Insemination Centers in Indonesia

Green tea extract increases the quality and reduced DNA mutation of post-thawed Kacang buck sperm

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