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A dual colour FISH method for routine validation of sexed Bos taurus semen

BACKGROUND: Usage of sexed semen that allows to choose the gender of the calves, is commonly practiced in livestock industry as a profitable breeding alternative, especially in dairy farming. The flow cytometric cell sorting is the only commercially available method for bovine sperm sexing. For vali...

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Autores principales: Reinsalu, Olavi, Scheler, Ott, Mikelsaar, Ruth, Mikelsaar, Aavo-Valdur, Hallap, Triin, Jaakma, Ülle, Padrik, Peeter, Kavak, Ants, Salumets, Andres, Kurg, Ants
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6446427/
https://www.ncbi.nlm.nih.gov/pubmed/30943959
http://dx.doi.org/10.1186/s12917-019-1839-3
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author Reinsalu, Olavi
Scheler, Ott
Mikelsaar, Ruth
Mikelsaar, Aavo-Valdur
Hallap, Triin
Jaakma, Ülle
Padrik, Peeter
Kavak, Ants
Salumets, Andres
Kurg, Ants
author_facet Reinsalu, Olavi
Scheler, Ott
Mikelsaar, Ruth
Mikelsaar, Aavo-Valdur
Hallap, Triin
Jaakma, Ülle
Padrik, Peeter
Kavak, Ants
Salumets, Andres
Kurg, Ants
author_sort Reinsalu, Olavi
collection PubMed
description BACKGROUND: Usage of sexed semen that allows to choose the gender of the calves, is commonly practiced in livestock industry as a profitable breeding alternative, especially in dairy farming. The flow cytometric cell sorting is the only commercially available method for bovine sperm sexing. For validation of the sexing procedure several methods have been developed including sperm fluorescence in situ hybridisation techniques. Latter usually include the use of pre-labelled nucleotides for probe synthesis which is relatively expensive approach compared to combined application of aminoallyl-dUTP and chemical binding of fluorescent dyes. Here a sex determining dual colour bovine sperm fluorescence in situ hybridisation method is presented which is considered more cost-effective technique than the previously reported approaches. RESULTS: The reliability of sex chromosome identifying probes, designed in silico, was proven on bovine metaphase plate chromosomes and through comparison with a commercially available standard method. In the dual colour FISH experiments of unsexed and sexed bovine sperm samples the hybridisation efficiency was at least 98%, whereas the determined sex ratios were not statistically different from the expected. Very few cells carried both of the sex chromosome-specific signals (less than 0.2%). CONCLUSIONS: A protocol for a dual colour bovine sperm FISH method is provided which is cost-effective, simple and fast for sex determination of spermatozoa in bull semen samples.
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spelling pubmed-64464272019-04-15 A dual colour FISH method for routine validation of sexed Bos taurus semen Reinsalu, Olavi Scheler, Ott Mikelsaar, Ruth Mikelsaar, Aavo-Valdur Hallap, Triin Jaakma, Ülle Padrik, Peeter Kavak, Ants Salumets, Andres Kurg, Ants BMC Vet Res Methodology Article BACKGROUND: Usage of sexed semen that allows to choose the gender of the calves, is commonly practiced in livestock industry as a profitable breeding alternative, especially in dairy farming. The flow cytometric cell sorting is the only commercially available method for bovine sperm sexing. For validation of the sexing procedure several methods have been developed including sperm fluorescence in situ hybridisation techniques. Latter usually include the use of pre-labelled nucleotides for probe synthesis which is relatively expensive approach compared to combined application of aminoallyl-dUTP and chemical binding of fluorescent dyes. Here a sex determining dual colour bovine sperm fluorescence in situ hybridisation method is presented which is considered more cost-effective technique than the previously reported approaches. RESULTS: The reliability of sex chromosome identifying probes, designed in silico, was proven on bovine metaphase plate chromosomes and through comparison with a commercially available standard method. In the dual colour FISH experiments of unsexed and sexed bovine sperm samples the hybridisation efficiency was at least 98%, whereas the determined sex ratios were not statistically different from the expected. Very few cells carried both of the sex chromosome-specific signals (less than 0.2%). CONCLUSIONS: A protocol for a dual colour bovine sperm FISH method is provided which is cost-effective, simple and fast for sex determination of spermatozoa in bull semen samples. BioMed Central 2019-04-03 /pmc/articles/PMC6446427/ /pubmed/30943959 http://dx.doi.org/10.1186/s12917-019-1839-3 Text en © The Author(s). 2019 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Methodology Article
Reinsalu, Olavi
Scheler, Ott
Mikelsaar, Ruth
Mikelsaar, Aavo-Valdur
Hallap, Triin
Jaakma, Ülle
Padrik, Peeter
Kavak, Ants
Salumets, Andres
Kurg, Ants
A dual colour FISH method for routine validation of sexed Bos taurus semen
title A dual colour FISH method for routine validation of sexed Bos taurus semen
title_full A dual colour FISH method for routine validation of sexed Bos taurus semen
title_fullStr A dual colour FISH method for routine validation of sexed Bos taurus semen
title_full_unstemmed A dual colour FISH method for routine validation of sexed Bos taurus semen
title_short A dual colour FISH method for routine validation of sexed Bos taurus semen
title_sort dual colour fish method for routine validation of sexed bos taurus semen
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6446427/
https://www.ncbi.nlm.nih.gov/pubmed/30943959
http://dx.doi.org/10.1186/s12917-019-1839-3
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