Effect of bovine sperm chromatin integrity evaluated using three different methods on in vitro fertility

Castro, Luiz Guilherme Martins; Siqueira, Adriano Felipe Perez; Hamilton, Tom R.; Mendes, Clayton Quirino; Visintin, J. A.; Assumpção, M. E. O. A.

doi:10.1016/j.theriogenology.2017.11.006

Cited by 30 publications

(21 citation statements)

References 59 publications

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“…Whereas alkaline comet is a technique used to quantify the grade of DNA fragmentation, it is complementary to analysis of sperm DNA susceptibility to denaturation. This relationship between sperm DNA susceptibility to denaturation and DNA fragmentation was also observed in cattle [ 36 , 37 ]. Sperm with higher susceptibility to DNA fragmentation would suffer chromatin damage quickly under oxidative stress condition [ 38 ].…”

Section: Discussionsupporting

confidence: 55%

Effect of Heat Stress on Sperm DNA: Protamine Assessment in Ram Spermatozoa and Testicle

Hamilton

Siqueira

Castro

et al. 2018

Oxidative Medicine and Cellular Longevity

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Sperm DNA fragmentation is considered one of the main causes of male infertility. The most accepted causes of sperm DNA damage are deleterious actions of reactive oxygen species (ROS), defects in protamination, and apoptosis. Ram sperm are highly prone to those damages due to the high susceptibility to ROS and to oxidative stress caused by heat stress. We aimed to evaluate the effects of heat stress on the chromatin of ejaculated and epididymal sperm and the activation of apoptotic pathways in different cell types in ram testis. We observed higher percentages of ejaculated sperm with increased chromatin fragmentation in the heat stress group; a fact that was unexpectedly not observed in epididymal sperm. Heat stress group presented a higher percentage of spermatozoa with DNA fragmentation and increased number of mRNA copies of transitional protein 1. Epididymal sperm presented greater gene expression of protamine 1 on the 30th day of the spermatic cycle; however, no differences in protamine protein levels were observed in ejaculated sperm and testis. Localization of proapoptotic protein BAX or BCL2 in testis was not different. In conclusion, testicular heat stress increases ram sperm DNA fragmentation without changes in protamination and apoptotic patterns.

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Section: Discussionsupporting

confidence: 55%

Effect of Heat Stress on Sperm DNA: Protamine Assessment in Ram Spermatozoa and Testicle

Hamilton

Siqueira

Castro

et al. 2018

Oxidative Medicine and Cellular Longevity

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“…Chromatin resistance analysis was performed according to previous work of our group [ 24 ]. Briefly, 375,000 of prepared cells were added to 50μL of TNE buffer (10mM Tris-HCl, 0.15M NaCl, 1mM EDTA disodium pH = 7.4) and 100μL of acid detergent solution (0.08M HCl, 0.15M NaCl, 0.1% (v/v) Triton X-100, pH = 1.4).…”

Section: Methodsmentioning

confidence: 99%

Sperm traits on in vitro production (IVP) of bovine embryos: Too much of anything is good for nothing

et al. 2018

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Sperm samples used on fertilization strongly influence the in vitro production (IVP) rates. However, sperm traits behind this effect are not stated consistently until now. This study aimed to evaluate the isolated and combined effect of some sperm traits (MB: total motility before Percoll® gradient, MA: total motility after Percoll® gradient, AI: acrosome integrity, MI: membrane integrity, MP: mitochondrial membrane potential, and CR: chromatin resistance) on IVP rates. This is the first study focusing on the isolated effect of distinct traits. For this purpose, the experiment was divided in three steps. In first step, to study behavior of traits sperm samples (n = 63 batches) were analyzed and ranked based on each trait. In second step, samples ranked were selected from target ranks regions and allocated in groups of four to five batches, creating Higher and Lower groups, according to two different approaches. One aimed to form groups that differed to all sperm traits simultaneously (effect of combined traits). The other aimed to form groups that differed only to a single sperm trait while no differences were observed for the remaining traits (effect of each isolated trait). In third step, for each group successfully formed in step 2, sperm samples were individually and prospectively used for IVP. Cleavage, embryo development and blastocyst rates were recorded and compared between Higher and Lower of respective trait groups. Surprisingly, evaluation of isolated effects revealed that lower levels of MB, AI and MP resulted in higher embryo development and blastocyst rates (p<0.05), which was not observed on cleavage rate. We conclude that sperm traits strongly influence embryo development after in vitro fertilization (IVF), affecting the zygote competence to achieve blastocyst stage. Individually, levels of MB, AI or MP could be some of the key traits that may define IVP efficiency on current systems of embryo production.

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“…Many publications have validated the use of this assessment in animals and humans (De Ambrogi et al, 2006;Dias et al, 2006;Garcia-Macias et al, 2006;Hernandez et al, 2006;Kasimanickam et al, 2006;Perez et al, 2014;Hamilton et al, 2016;Castro et al, 2018); and only sperm chromatin integrity assay has an exact standardization (reviewed by Evenson et al, 2016). In our experience, no differences were observed in the percentage of positive sperm for AO between bulls with high and low fertility in IVF (Castro et al, 2018). However, ram sperm with high susceptibility to induced lipid peroxidation presented a high percentage of native positive cells stained with AO, indicating higher fragility of sperm DNA (Hamilton et al, 2016).…”

Section: Assessment Of Sperm Dna Integritymentioning

confidence: 74%

Sperm DNA fragmentation: causes and identification

Hamilton

Assumpção

2019

Zygote

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SummarySperm DNA fragmentation is referred to as one of the main causes of male infertility. Failures in the protamination process, apoptosis and action of reactive oxygen species (ROS) are considered the most important causes of DNA fragmentation. Action of ROS or changes in sperm protamination would increase the susceptibility of sperm DNA to fragmentation. Routine semen analysis is unable to estimate sperm chromatin damage. Sperm DNA integrity influences sperm functional capability, therefore tests that measure sperm DNA fragmentation are important to assess fertility disorders. Actually, there is a considerable number of methods for assessing sperm DNA fragmentation and chromatin integrity, sperm chromatin stability assay (SCSA modified), sperm chromatin dispersion (SCD), comet assay, transferase dUTP nick end labelling (TUNEL); and protamine evaluation in sperm chromatin assay, such as toluidine blue, CMA3, protamine expression and evaluation of cysteine radicals. This review aims to describe the main causes of sperm DNA fragmentation and the tests commonly used to evaluate sperm DNA fragmentation.

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Effect of bovine sperm chromatin integrity evaluated using three different methods on in vitro fertility

Cited by 30 publications

References 59 publications

Effect of Heat Stress on Sperm DNA: Protamine Assessment in Ram Spermatozoa and Testicle

Effect of Heat Stress on Sperm DNA: Protamine Assessment in Ram Spermatozoa and Testicle

Sperm traits on in vitro production (IVP) of bovine embryos: Too much of anything is good for nothing

Sperm DNA fragmentation: causes and identification

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