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Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation

BACKGROUND: The use of flow cytometry (FC) to evaluate sperm DNA fragmentation via deoxynucleotidyl transferase terminal fluorescein dUTP nick-end labeling (TUNEL) has shown inconsistencies compared with conventional fluorescent microscopic analyses. It has been hypothesized that the observed discre...

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Autores principales: Sadeghi, Niloofar, Tavalaee, Marziyeh, Kiani-Esfahani, Abbas, Moazamian, Aron, Gharagozloo, Parviz, Aitken, Robert J., Drevet, Joël R., Nasr-Esfahani, Mohammad Hossein
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8529777/
https://www.ncbi.nlm.nih.gov/pubmed/34670490
http://dx.doi.org/10.1186/s12610-021-00143-7
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author Sadeghi, Niloofar
Tavalaee, Marziyeh
Kiani-Esfahani, Abbas
Moazamian, Aron
Gharagozloo, Parviz
Aitken, Robert J.
Drevet, Joël R.
Nasr-Esfahani, Mohammad Hossein
author_facet Sadeghi, Niloofar
Tavalaee, Marziyeh
Kiani-Esfahani, Abbas
Moazamian, Aron
Gharagozloo, Parviz
Aitken, Robert J.
Drevet, Joël R.
Nasr-Esfahani, Mohammad Hossein
author_sort Sadeghi, Niloofar
collection PubMed
description BACKGROUND: The use of flow cytometry (FC) to evaluate sperm DNA fragmentation via deoxynucleotidyl transferase terminal fluorescein dUTP nick-end labeling (TUNEL) has shown inconsistencies compared with conventional fluorescent microscopic analyses. It has been hypothesized that the observed discrepancies could be attributed to the presence of apoptotic bodies that can be labeled with merocyanine 540, the so-called M540 bodies. In order to verify this hypothesis and determine the accuracy of our in-house FC-assisted evaluation of spermatozoa parameters, we used FC to evaluate both the fragmentation of sperm DNA using the TUNEL assay and the oxidation of sperm DNA using the 8-OHdG assay on semen samples with or without M540 bodies. RESULTS: We show that the presence of M540 bodies lead to underestimation of both the level of sperm DNA fragmentation and sperm DNA oxidation when using FC assisted detection systems. We also observed that this situation is particularly pertinent in semen samples classified as abnormal with respect to the routine WHO semen evaluation as they appear to contain more M540 bodies than normal samples. CONCLUSIONS: We conclude that M540 bodies interfere with both FC-conducted assays designed to evaluate sperm nuclear/DNA integrity. Exclusion of these contaminants in unprepared semen samples should be performed in order to correctly appreciate the true level of sperm DNA/nuclear damage which is known to be a critical male factor for reproductive success. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12610-021-00143-7.
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spelling pubmed-85297772021-10-25 Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation Sadeghi, Niloofar Tavalaee, Marziyeh Kiani-Esfahani, Abbas Moazamian, Aron Gharagozloo, Parviz Aitken, Robert J. Drevet, Joël R. Nasr-Esfahani, Mohammad Hossein Basic Clin Androl Research Article BACKGROUND: The use of flow cytometry (FC) to evaluate sperm DNA fragmentation via deoxynucleotidyl transferase terminal fluorescein dUTP nick-end labeling (TUNEL) has shown inconsistencies compared with conventional fluorescent microscopic analyses. It has been hypothesized that the observed discrepancies could be attributed to the presence of apoptotic bodies that can be labeled with merocyanine 540, the so-called M540 bodies. In order to verify this hypothesis and determine the accuracy of our in-house FC-assisted evaluation of spermatozoa parameters, we used FC to evaluate both the fragmentation of sperm DNA using the TUNEL assay and the oxidation of sperm DNA using the 8-OHdG assay on semen samples with or without M540 bodies. RESULTS: We show that the presence of M540 bodies lead to underestimation of both the level of sperm DNA fragmentation and sperm DNA oxidation when using FC assisted detection systems. We also observed that this situation is particularly pertinent in semen samples classified as abnormal with respect to the routine WHO semen evaluation as they appear to contain more M540 bodies than normal samples. CONCLUSIONS: We conclude that M540 bodies interfere with both FC-conducted assays designed to evaluate sperm nuclear/DNA integrity. Exclusion of these contaminants in unprepared semen samples should be performed in order to correctly appreciate the true level of sperm DNA/nuclear damage which is known to be a critical male factor for reproductive success. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12610-021-00143-7. BioMed Central 2021-10-21 /pmc/articles/PMC8529777/ /pubmed/34670490 http://dx.doi.org/10.1186/s12610-021-00143-7 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Sadeghi, Niloofar
Tavalaee, Marziyeh
Kiani-Esfahani, Abbas
Moazamian, Aron
Gharagozloo, Parviz
Aitken, Robert J.
Drevet, Joël R.
Nasr-Esfahani, Mohammad Hossein
Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation
title Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation
title_full Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation
title_fullStr Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation
title_full_unstemmed Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation
title_short Apoptotic M540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm DNA fragmentation and oxidation
title_sort apoptotic m540 bodies present in human semen interfere with flow cytometry-assisted assessment of sperm dna fragmentation and oxidation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8529777/
https://www.ncbi.nlm.nih.gov/pubmed/34670490
http://dx.doi.org/10.1186/s12610-021-00143-7
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