Abundant alkali-sensitive sites in DNA of human and mouse sperm

Singh, Narendra P.; Danner, David B.; Tice, Raymond R.; McCoy, Michael T.; Collins, Gary D.; Schneider, Edward L.

doi:10.1016/0014-4827(89)90344-3

Cited by 238 publications

(131 citation statements)

References 16 publications

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“…A study using a pH 13 alkaline COMET assay showed that 20% of the total DNA fluorescence was in the comet tail of untreated spermatozoa, which is significantly higher than the 5% characteristic basal level of breaks in somatic cells (Hughes et al, 1996;Singh and Stephens, 1998). High basal levels of strand breaks were found in normozoospermic fertile men, indicating that normal human sperm chromatin has a high concentration of single-strand breaks that are alkali labile but not necessarily associated with reduced fertility (Singh et al, 1989). The presence of alkali labile DNA fragments in COMET assays could decrease the sensitivity of the assay to DNA fragments resulting from doublestrand breaks that are indicative of DNA damage and infertility.…”

Section: Comet Assaymentioning

confidence: 99%

Sperm Chromatin Structure Assay: Its Clinical Use for Detecting Sperm DNA Fragmentation in Male Infertility and Comparisons With Other Techniques

Evenson

Larson

Jost

2002

Journal of Andrology

894

768

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The Sperm Chromatin Structure Assay (SCSA) was first described in the December 5, 1980 issue of Science (Evenson et al, 1980a). The data described in that article showed a significant difference between proven fertile and subfertility or infertility in men and bulls and the susceptibility to denaturation of their sperm nuclear DNA. A subsequent study (Evenson et al, 1984) described SCSA data obtained from men with testicular cancer. The data were heterogeneous, and begged the question, what was SCSA measuring? Great effort has been spent over the past 15 years on animal model systems, mostly related to dose-response toxicology experiments and large animal fertility trials. The results from these studies showed that SCSA was highly dose-responsive to toxicants, highly repeatable, and provided meaningful biological information on sperm nuclear DNA defects. The extensive data from nonhumans (Evenson et al, 1985(Evenson et al, , 1989a(Evenson et al, ,b, 1993a(Evenson et al, ,b,c, 1994Ballachey et al, 1987Ballachey et al, , 1988Evenson and Jost, 1993; Sailer et al, 1995a,b;Evenson, 1999a), and human toxicology and fertility studies (Evenson et al, 1978(Evenson et al, , 1980a(Evenson et al, ,b, 1984(Evenson et al, , 1991Evenson and Melamed, 1983;Evenson, 1999b;Larson et al, 1999Larson et al, , 2000Larson et al, , 2001 provide This work was supported in part by grant R827019 from the US Environmental Protection Agency, grants EPS-9720642 and OSR-9452894 from the National Science Foundation, and by SCSA Diagnostics, Inc. This is South Dakota Agricultural Experiment Station publication 3273 of the journal series.Correspondence to: Donald P. Evenson, Olson Biochemistry Laboratories, ASC 136, South Dakota State University, Brookings, SD 57006 (e-mail: donald evenson@sdstate.edu).Received for publication September 20, 2001; accepted for publication September 20, 2001. compelling evidence that SCSA will be useful in clinical human semen analysis (Figure 1).After reviewing current infertility center sites on the World Wide Web and approximately a dozen lay-oriented books on infertility and assisted reproductive technology (ART), it was amazing that not a single reference contained any information about the negative influence of fragmented sperm nuclear DNA on successful pregnancy outcome. Furthermore, few physicians are aware of scientific literature on this subject. Currently, if a spermatozoon has some motility as seen under a light microscope, and reasonable morphology, then many ART clinics assume that the sperm DNA must be fine. However, a number of cases have been documented in which couples have had multiple failed attempts at intracytoplasmic sperm injection (ICSI), even using donor eggs, and we found that the fragmentation of sperm DNA was above our threshold for less than 1% probability of achieving a successful pregnancy. Spermatozoa with defective DNA can fertilize an oocyte, produce high-quality early stage embryos, and then, in relationship to the extent of DNA damage, fail in producing a successful term ...

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Section: Comet Assaymentioning

confidence: 99%

Sperm Chromatin Structure Assay: Its Clinical Use for Detecting Sperm DNA Fragmentation in Male Infertility and Comparisons With Other Techniques

Evenson

Larson

Jost

2002

Journal of Andrology

894

768

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“…The alkaline variant of the comet assay assesses both the single and double strand breaks and the alkali labile sites as compared to the neutral comet assay which measures only DSB's. The alkaline comet assay overestimate true DNA strand breakage in spermatozoa because of artificial damage induced at alkali-labile sites within the DNA strand [105]. Regarding alkaline and neutral variant of comet assay different workers have varied opinion in deciding which is better.…”

Section: Research Laboratory Aspects and Relevance Of Dna Integrity Tmentioning

confidence: 99%

Sperm DNA integrity assays: diagnostic and prognostic challenges and implications in management of infertility

Shamsi

Imam

Dada

2011

J Assist Reprod Genet

130

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Sperm is not a simple carrier of paternal genetic information but its role extends clearly beyond fertilization. Integrity of sperm genome is an essential pre-requisite for birth of healthy offspring and evaluation of sperm should entail DNA integrity analysis. DNA integrity analysis is a better diagnostic and prognostic marker of sperm reproductive potential. Conventional semen analysis emphasizes on sperm concentration, viability, motility and morphology and has been proven to be a poor indicator of reproductive potential and pregnancy outcome. To overcome the drawbacks associated with conventional semen analysis more useful fertility tests and molecular biomarkers have been explored. Among the different tests which have evolved for assessing the sperm reproductive potential, tests for sperm DNA quality are most promising. Sperm DNA damage has been closely associated with numerous indicators of reproductive health including fertilization, embryo quality, implantation, spontaneous abortion, congenital malformations and childhood diseases. It therefore has great potential as a prognostic test for both in vitro and in vivo conception. This review presents an updated account of tests that have better diagnostic and prognostic implications in the evaluation of sperm DNA damage. The basic principles, outline of methodology, advantage, disadvantage, clinical significance of each technique and implications of these tests have been discussed. The logistics of each test with respect to available resources and equipment in an andrology laboratory, the feasibility of performing these tests in routine diagnostic workup of infertile men and the opportunities and challenges provided by DNA testing in male fertility determination are also presented.

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“…To date, no sensitive highly quantitative method has been used to examine the relationship between subject's age and sperm DNA damage. Methods such as filter elution assays (19 -21), terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) (22), comet assay, or microgel electrophoresis technique (23)(24)(25)(26)(27)(28)(29)(30)(31)(32) have been used to quantify DNA damage in sperm cells. The neutral microgel electrophoresis technique is a sensitive and simple method of evaluating double-strand break DNA damage in individual sperm cells (24,(33)(34)(35).…”

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confidence: 99%