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Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay

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Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay. / Pollock, K; Gosálvez, J; Arroyo, F; López-Fernández, C; Guille, M; Noble, A; Johnston, S D.

In: Reproduction, Fertility and Development, Vol. 27, No. 8, 03.06.2014, p. 1168-1174.

Research output: Contribution to journalArticlepeer-review

Harvard

Pollock, K, Gosálvez, J, Arroyo, F, López-Fernández, C, Guille, M, Noble, A & Johnston, SD 2014, 'Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay', Reproduction, Fertility and Development, vol. 27, no. 8, pp. 1168-1174. https://doi.org/10.1071/RD14070

APA

Pollock, K., Gosálvez, J., Arroyo, F., López-Fernández, C., Guille, M., Noble, A., & Johnston, S. D. (2014). Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay. Reproduction, Fertility and Development, 27(8), 1168-1174. https://doi.org/10.1071/RD14070

Vancouver

Pollock K, Gosálvez J, Arroyo F, López-Fernández C, Guille M, Noble A et al. Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay. Reproduction, Fertility and Development. 2014 Jun 3;27(8):1168-1174. https://doi.org/10.1071/RD14070

Author

Pollock, K ; Gosálvez, J ; Arroyo, F ; López-Fernández, C ; Guille, M ; Noble, A ; Johnston, S D. / Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay. In: Reproduction, Fertility and Development. 2014 ; Vol. 27, No. 8. pp. 1168-1174.

Bibtex

@article{02083538c17e4f299d40ef4e073316cf,
title = "Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay",
abstract = "The integrity of sperm DNA is becoming increasingly recognised as an important parameter of semen quality, but there are no published reports of this procedure for any amphibian. The primary aim of this study was to apply a modified sperm chromatin dispersion (SCD) test (Halomax) to an amphibian sperm model (African clawed frog; Xenopus laevis) and to validate the assay against in situ nick translation (ISNT) and the double-comet assay procedure. Inactivated spermatozoa were collected from fresh testes (n=3). Sperm DNA fragmentation (SDF) for each sperm sample was conducted immediately following activation (T0) and again after 1h (T1) and 24h (T24) of incubation at room temperature in order to produce a range of spermatozoa with differing levels of DNA damage. The SCD procedure resulted in the production of three nuclear morphotypes; amphibian sperm morphotype 1 (ASM-1) and ASM-2 showed no evidence of DNA damage, whereas ASM-3 spermatozoa were highly fragmented with large halos of dispersed DNA fragments and a reduced nuclear core. ISNT confirmed that ASM-3 nuclei contained damaged DNA. There was a significant correlation (r=0.9613) between the levels of ASM-3 detected by the SCD test and SDF revealed by the double-comet assay.",
author = "K Pollock and J Gos{\'a}lvez and F Arroyo and C L{\'o}pez-Fern{\'a}ndez and M Guille and A Noble and Johnston, {S D}",
year = "2014",
month = jun,
day = "3",
doi = "10.1071/RD14070",
language = "English",
volume = "27",
pages = "1168--1174",
journal = "Reproduction, Fertility and Development",
issn = "1031-3613",
publisher = "CSIRO",
number = "8",

}

RIS

TY - JOUR

T1 - Validation of the sperm chromatin dispersion (SCD) test in the Amphibian Xenopus laevis using in situ nick translation and comet assay

AU - Pollock, K

AU - Gosálvez, J

AU - Arroyo, F

AU - López-Fernández, C

AU - Guille, M

AU - Noble, A

AU - Johnston, S D

PY - 2014/6/3

Y1 - 2014/6/3

N2 - The integrity of sperm DNA is becoming increasingly recognised as an important parameter of semen quality, but there are no published reports of this procedure for any amphibian. The primary aim of this study was to apply a modified sperm chromatin dispersion (SCD) test (Halomax) to an amphibian sperm model (African clawed frog; Xenopus laevis) and to validate the assay against in situ nick translation (ISNT) and the double-comet assay procedure. Inactivated spermatozoa were collected from fresh testes (n=3). Sperm DNA fragmentation (SDF) for each sperm sample was conducted immediately following activation (T0) and again after 1h (T1) and 24h (T24) of incubation at room temperature in order to produce a range of spermatozoa with differing levels of DNA damage. The SCD procedure resulted in the production of three nuclear morphotypes; amphibian sperm morphotype 1 (ASM-1) and ASM-2 showed no evidence of DNA damage, whereas ASM-3 spermatozoa were highly fragmented with large halos of dispersed DNA fragments and a reduced nuclear core. ISNT confirmed that ASM-3 nuclei contained damaged DNA. There was a significant correlation (r=0.9613) between the levels of ASM-3 detected by the SCD test and SDF revealed by the double-comet assay.

AB - The integrity of sperm DNA is becoming increasingly recognised as an important parameter of semen quality, but there are no published reports of this procedure for any amphibian. The primary aim of this study was to apply a modified sperm chromatin dispersion (SCD) test (Halomax) to an amphibian sperm model (African clawed frog; Xenopus laevis) and to validate the assay against in situ nick translation (ISNT) and the double-comet assay procedure. Inactivated spermatozoa were collected from fresh testes (n=3). Sperm DNA fragmentation (SDF) for each sperm sample was conducted immediately following activation (T0) and again after 1h (T1) and 24h (T24) of incubation at room temperature in order to produce a range of spermatozoa with differing levels of DNA damage. The SCD procedure resulted in the production of three nuclear morphotypes; amphibian sperm morphotype 1 (ASM-1) and ASM-2 showed no evidence of DNA damage, whereas ASM-3 spermatozoa were highly fragmented with large halos of dispersed DNA fragments and a reduced nuclear core. ISNT confirmed that ASM-3 nuclei contained damaged DNA. There was a significant correlation (r=0.9613) between the levels of ASM-3 detected by the SCD test and SDF revealed by the double-comet assay.

U2 - 10.1071/RD14070

DO - 10.1071/RD14070

M3 - Article

C2 - 25482041

VL - 27

SP - 1168

EP - 1174

JO - Reproduction, Fertility and Development

JF - Reproduction, Fertility and Development

SN - 1031-3613

IS - 8

ER -

ID: 2552250