Evaluation of Chromosome Breakage and DNA Integrity in Sperm: An Investigation of Remote Semen Collection Conditions

Young, Karen E.; Robbins, Wendie A.; Lin, Xiao; Elashoff, David; Rothmann, Susan A.; Perreault, Sally D.

doi:10.1002/j.1939-4640.2003.tb03136.x

Cited by 40 publications

(23 citation statements)

References 38 publications

(49 reference statements)

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“…These incorporated labeled nucleotides can be detected in spermatozoa by FC, fluorescence microscopy or light microscopy. 96 TUNEL can simultaneously detect single-and double-strand breaks. By TUNEL the degree of DNA damage within a cell cannot be quantified, which only reveals the number of cells within a population with DNA damage An advantage of the TUNEL assay is its application in FC, 97 although Domínguez-Fandos et al 98 found 2.6 times greater sperm damage by FC than that of fluorescent microscopy, thus suggesting that it may overestimate damage.…”

Section: Sperm Intactnessmentioning

confidence: 99%

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Hossain¹,

Johannisson²,

Wallgren³

et al. 2011

Asian J Androl

140

120

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Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of 'bench-top' flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness when exposed to exogenous stimuli, and to increase our capabilities to sort spermatozoa for potential fertilizing capacity, or chromosomal sex. The present review summarizes the state of the art regarding flow cytometry applied to animal andrology, albeit keeping an open comparative intent. It critically evaluates the present and future capabilities of flow cytometry for the diagnostics of potential fertility and for the development of current reproductive technologies such as sperm freezing, sperm selection and sperm sorting. The flow cytometry methods will probably further revolutionize our understanding of the sperm physiology and their functionality, and will undoubtedly extend its application in isolating many uncharacterized features of spermatozoa. However, continuous follow-up of the methods is a necessity owing to technical developments and the complexity of mapping spermatozoa.

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Section: Sperm Intactnessmentioning

confidence: 99%

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Hossain¹,

Johannisson²,

Wallgren³

et al. 2011

Asian J Androl

140

120

View full text Add to dashboard Cite

show abstract

“…Some of these variants are widely utilized, for instance the use of formaldehyde to fix semen samples or the use of fluorescence microscopy (the most available technology to reveal fluorescence) to score samples. However, differences in the TUNEL assay include: (i) preparation, fixation, and permeabilization of samples, (ii) protocols for labeling DNA breaks, (iii) technologies to detect fluorescence, and (iv) methods to analyze flow cytometric data (Table I) [Young et al 2003;Zhang et al 2008], to promote nuclei decondensation.…”

Section: Existing Variants Of the Tunel Procedures Greatly Affect Measmentioning

confidence: 99%

“…By reviewing studies conducted in raw semen from similar groups Muciaccia et al 2007;O'Flaherty et al 2008;Cohen-Bacrie et al 2009;Tunc et al 2008;Martin et al 2005Formaldeyde d Muratori et al 2003Caglar et al 2007;Tarozzi et al 2009;Domínguez-Fandos et al 2007;Young et al 2003;Avendañ o et al 2009a and2009b;Stronati et al 2006;Ramos et al 2008;Tesarik et al 2004 Permeabilization with detergents Absent Martin et al 2005;O'Flaherty et al 2008;Young et al 2003;Said et al 2006Present e Chohan et al 2006Aoki et al 2006;Caglar et al 2007;Varum et al 2007;Muratori et al 2008a andTunc et al 2008;Frydman et al 2008;Avendañ o et al 2009a and2009b Labelling Direct labeling f O'Flaherty et al 2008;Zhang et al 2008;Aoki et al 2006;Torregrosa et al 2006;Young et al 2003;Muratori et al 2008a;.Greco et al 2005;Paasch et al 2004;Brugnon et al 2006;Donnelly et al 2000 Indirect labeling g Varum et al 2007;Sergerie et al 2005;P...…”

Section: Comparing Flow Cytometry and Fluorescence Microscopy For Spementioning

confidence: 99%

Critical Aspects of Detection of Sperm DNA Fragmentation by Tunel/Flow Cytometry

Muratori

Tamburrino

Marchiani

et al. 2010

Systems Biology in Reproductive Medicine

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Despite the many studies documenting an increase of sperm DNA damage in subfertile and infertile subjects, determining whether this parameter is relevant for the clinic is still an open question. Indeed, results of clinical investigations on sperm DNA damage and outcome of ART procedures are often conflicting as many factors affect the predictive power of these tests. The techniques used to reveal such damage is one of these factors. Techniques detecting sperm DNA damage are indeed numerous and heterogeneous. In addition, it is not obvious that they reveal the same type of DNA damage and that their results are equivalent. One of the available methods to detect sperm DNA damage is the TUNEL assay. Since it was developed in the 1990s the TUNEL assay has been widely employed, becoming one of the most popular strategies to investigate the origin, the mechanism, and the clinical meaning of sperm DNA damage. Our group has used TUNEL coupled to flow cytometry to investigate sperm DNA fragmentation for about 10 years. According to our experience, this technique presents some pitfalls and limitations in the accuracy and reproducibility of the measures of sperm DNA fragmentation. In this review, we discuss several technical features of TUNEL and report the solutions adopted by our group to overcome some of its limitations.

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“…Transport or storage of liquid human semen overnight, cooled or at room temperature after extension in one of several buffers, including those formulated with hen's egg yolk, protects sperm motility and function [18,[21][22][23][24][25][26][27][28]. According to Manjunath [29], the protective effect of hen's egg yolk or milk-based extenders appears to be due to low density lipoprotein (LDL) components in the buffer that competitively bind with and scavenge a family of proteins known, in hoof stock, as binder of sperm (BSP) proteins, reducing the sensitivity of the sperm membrane to cooling, and protecting/ replacing cholesterol within the sperm membrane.…”

Section: Discussionmentioning

confidence: 99%

Increased count, motility, and total motile sperm cells collected across three consecutive ejaculations within 24 h of oocyte retrieval: implications for management of men presenting with low numbers of motile sperm for assisted reproduction

Said

Reed

2015

J Assist Reprod Genet

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Purpose The purpose of this study was to quantitate changes in seminal volume, sperm count, motility, qualitative forward progression, and total motile sperm cells per ejaculate, across three consecutive ejaculates collected from individuals within 24 h preceding an IVF cycle. Methods Men presenting with oligoasthenozoospermia or asthenozoospemia attempted three ejaculates within 24 h preceding IVF. Ejaculate 1 was produced the afternoon prior to oocyte retrieval, and ejaculates 2 and 3 were produced the morning of oocyte retrieval with 2-3 h between collections. Ejaculates 1 and 2 were extended 1:1 v/v with room temperature rTYBS. Test tubes were placed into a beaker of room temperature water, then placed at 4°C for gradual cooling. Ejaculate 3 was not extended, but pooled with ejaculates 1 and 2 and processed for intracytoplasmic sperm injection (ICSI). Results Out of 109 oocyte retrievals, 28 men were asked to attempt multiple consecutive ejaculations. Among this population, 25/28 (89.3 %) were successful, and 3/28 men (10.7 %) could only produce two ejaculates. Mean volumes for ejaculates 1, 2, and 3 were significantly different from each other (p<0.01); the volume decreased for each ejaculate. Mean sperm counts, motility, qualitative forward progression, and total motile cells per ejaculate for the ejaculates1, 2, and 3 demonstrated the following: ejaculates 2 and 3 were not significantly different, but counts, motility, and total motile sperm were improved over ejaculate 1 (p<0.01). Conclusions Pooling three consecutive ejaculates within 24 h increased the numbers of available motile sperm in this population by 8-fold compared to the first ejaculate alone, facilitating avoidance of sperm cryopreservation and additional centrifugation steps that could affect sperm viability and/or function.

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Evaluation of Chromosome Breakage and DNA Integrity in Sperm: An Investigation of Remote Semen Collection Conditions

Cited by 40 publications

References 38 publications

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Critical Aspects of Detection of Sperm DNA Fragmentation by Tunel/Flow Cytometry

Increased count, motility, and total motile sperm cells collected across three consecutive ejaculations within 24 h of oocyte retrieval: implications for management of men presenting with low numbers of motile sperm for assisted reproduction

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