Loss of livestock breeding efficiency due to uncompensable sperm nuclear defects

1999 ◽  
Vol 11 (1) ◽  
pp. 1 ◽  
Author(s):  
D. P. Evenson
Keyword(s):  
Chromatin Structure ◽  
Male Fertility ◽  
Nuclear Dna ◽  
Early Embryo ◽  
Sperm Chromatin ◽  
Sperm Chromatin Structure Assay ◽  
Fertility Potential ◽  
Post Stress

An important goal of modern analyses of semen is to elucidate the molecular traits of mammalian sperm chromatin structural abnormalities, defined here as ‘uncompensable’, that lead to abnormalities in fertility, pronuclear formation, early embryo quality and pregnancy outcome. Sperm with uncompensable nuclear abnormalities are able to fertilize oocytes both in vivo and in vitro; however, due to the uncompensable trait(s), the embryo development may be abnormal. Uncompensable nuclear traits can be experimentally induced in bull sperm by a mild thermal insult to the testis. Sperm nuclear morphology abnormalities seen in ejaculates 11-days post stress are likely related to molecular changes in chromatin observed 3-days post stress by the flow cytometric sperm chromatin structure assay (SCSA). The SCSA measures the susceptibility of sperm nuclear DNA to denaturation in situ. This susceptibility has been correlated with the presence of DNA strand breaks that may be derived in part by oxidative stress and possibly by a unique, abortive apoptotic mechanism. The extent of DNA denaturation is not significantly related to the level of disulfide bonding between the chromatin protamines. The use of human sperm with uncompensable nuclear traits for artificial reproductive techniques is also discussed. The goal of this research is to remove from semen doses those sperm with uncompensable nuclear traits and thereby increase male fertility potential. Extra key words: male fertility potential, sperm chromatin structure assay (SCSA).

Evaluation of fertility potential by toluidine blue test and the sperm chromatin structure assay

2007 ◽  
Vol 88 ◽  
pp. S301
Author(s):  
R. Mahfouz ◽  
A. Agarwal ◽  
T.M. Said ◽  
J. Erenpreiss ◽  
A. Giwercman ◽  
...  
Keyword(s):  
Toluidine Blue ◽  
Chromatin Structure ◽  
Sperm Chromatin ◽  
Sperm Chromatin Structure Assay ◽  
Fertility Potential

scholarly journals Sperm chromatin structure assay versus sperm chromatin dispersion kits: Technical repeatability and choice of assisted reproductive technology procedure

2020 ◽  
Vol 47 (4) ◽  
pp. 277-283
Author(s):  
Vidya Laxme B ◽  
Silviya Stephen ◽  
Ramyashree Devaraj ◽  
Sridurga Mithraprabhu ◽  
Ricardo P. Bertolla ◽  
...  
Keyword(s):  
Assisted Reproductive Technology ◽  
Chromatin Structure ◽  
Reproductive Technology ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Vitro Fertilization ◽  
Sperm Chromatin Structure Assay ◽  
Sperm Dna ◽  
The Impact

Objective: The sperm DNA fragmentation index (DFI) guides the clinician’s choice of an appropriate assisted reproductive technology (ART) procedure. The DFI can be determined using commercially available methodologies, including sperm chromatin dispersion (SCD) kits and sperm chromatin structure assay (SCSA). Currently, when DFI is evaluated using SCD kits, the result is analyzed in reference to the SCSA-derived threshold for the choice of an ART procedure. In this study, we compared DFI values obtained using SCSA with those obtained using SCD and determined whether the difference affects the choice of ART procedure.Methods: We compared SCSA to two SCD kits, CANfrag (n=36) and Halosperm (n=31), to assess the DFI values obtained, the correlations between tests, the technical repeatability, and the impact of DFI on the choice of ART. Results: We obtained higher median DFI values using SCD kits than when using SCSA, and this difference was significant for the CANfrag kit (p<0.001). The SCD kits had significantly higher coefficients of variation than SCSA (p<0.0001). In vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) would be chosen for a significantly higher proportion of patients if a decision were made based on DFI derived from SCD rather than DFI determined using SCSA (p=0.003). Conclusion: Our results indicate that SCD kit-specific thresholds should be established in order to avoid the unnecessary use of IVF/ICSI based on sperm DNA damage for the management of infertility. Appropriate measures should be taken to mitigate the increased variability inherent to the methods used in these tests.

scholarly journals Evaluation of male fertility potential by Toluidine Blue test for sperm chromatin structure assessment

2009 ◽  
Vol 24 (7) ◽  
pp. 1569-1574 ◽  
Author(s):  
I. Tsarev ◽  
M. Bungum ◽  
A. Giwercman ◽  
J. Erenpreisa ◽  
T. Ebessen ◽  
...  
Keyword(s):  
Toluidine Blue ◽  
Chromatin Structure ◽  
Male Fertility ◽  
Sperm Chromatin ◽  
Structure Assessment ◽  
Fertility Potential

Psoralen photocrosslinking, a tool to study the chromatin structure of RNA polymerase I - transcribed ribosomal genes

2005 ◽  
Vol 83 (4) ◽  
pp. 449-459 ◽  
Author(s):  
Martin Toussaint ◽  
Geneviève Levasseur ◽  
Maxime Tremblay ◽  
Michel Paquette ◽  
Antonio Conconi
Keyword(s):  
Rna Polymerase ◽  
Chromatin Structure ◽  
Nuclear Dna ◽  
Rdna Locus ◽  
Ribosomal Genes ◽  
Rna Polymerase I ◽  
Coding Regions ◽  
Polymerase I

The chromatin structure of RNA polymerase I - transcribed ribosomal DNA (rDNA) is well characterized. In most organisms, i.e., lower eukaryotes, plants, and animals, only a fraction of ribosomal genes are transcriptionally active. At the chromatin level inactive rDNA is assembled into arrays of nucleosomes, whereas transcriptionally active rDNA does not contain canonical nucleosomes. To separate inactive (nucleosomal) and active (non-nucleosomal) rDNA, the technique of psoralen photocrosslinking has been used successfully both in vitro and in vivo. In Saccharomyces cerevisiae, the structure of rDNA chromatin has been particularly well studied during transcription and during DNA replication. Thus, the yeast rDNA locus has become a good model system to study the interplay of all nuclear DNA processes and chromatin. In this review we focused on the studies of chromatin in ribosomal genes and how these results have helped to address the fundamental question: What is the structure of chromatin in the coding regions of genes?Key words: active chromatin, FACT, lexosome, psoralen, photo-crosslinking, rDNA, RNA polymerase I.

228 BOAR FERTILITY AND SPERM CHROMATIN STRUCTURE ASSAY DEFINED SPERM DNA FRAGMENTATION

2009 ◽  
Vol 21 (1) ◽  
pp. 212 ◽  
Author(s):  
D. P. Evenson ◽  
K. Kasperson ◽  
R. L. Wixon ◽  
B. A. Didion
Keyword(s):  
Dna Fragmentation ◽  
Chromatin Structure ◽  
Canonical Correlation ◽  
Structural Integrity ◽  
Nuclear Dna ◽  
Sperm Chromatin ◽  
Sperm Dna Fragmentation ◽  
Sperm Chromatin Structure Assay ◽  
Sperm Dna ◽  
Dna Strand

The sperm chromatin structure assay (SCSA) was used retrospectively to characterize sperm from 18 sexually mature boars having fertility information. Boar fertility was defined by farrowing rate (FR) and average total number of pigs born (ANB) per litter of gilts and sows mated to individual boars. The SCSA uses flow cytometry to evaluate the structural integrity of sperm nuclear DNA. In brief, aliquots of frozen/thawed semen were treated for 30 s with a pH 1.2 buffer that denatures sperm DNA at the sites of DNA strand breaks. The samples are then stained with acridine orange (AO) that intercalates into native, double-stranded DNA and fluoresces green under the flow cytometer laser beam. Broken and denatured single-stranded DNA associates with AO and then collapses, causing a metachromatic shift from green to red fluorescence. Five thousand sperm are measured per sample at rates of ~200 s–1, thus giving the data strong statistical robustness. This fact coupled with the precision of the AO/DNA biochemistry provides a CV ~2% between replicate samples. The SCSA parameters measured in this study were the %DNA fragmentation index (%DFI; percentage of sperm with fragmented DNA) and standard deviation of DFI (SD DFI). The %DFI and SD DFI showed the following significant negative correlations with FR and ANB; %DFI v. FR r = –0.55, P < 0.01, SD DFI v. FR r = –0.67, P < 0.002, %DFI v. ANB r = –0.54, P < 0.01, and SD DFI v. ANB r = –0.54, P < 0.02. A discrimination analysis was used to distinguish between the boar groups above and below 6% DFI. The variables most useful to discriminate between 2 groups were SD DFI, FR, and ANB. The average squared canonical correlation for SD DFI was 0.82, P < 0.0001. The average squared canonical correlation for FR and ANB was 0.45, P < 0.003 and 0.54, P < 0.001, respectively. Odds ratios for %DFI and SD DFI were 1.5 times (P = 0.0003, CI 1.22, 1.94) and 2.5 times (P = 0.0001, CI 1.87, 3.32), respectively.

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