Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos

Abstract Nuclear/chromosomal integrity is an important prerequisite for the assessment of embryo quality in artificial reproductive technology. However, lipid-rich dark cytoplasm in bovine embryos prevents its observation by visible light microscopy. We performed live-cell imaging using confocal las...

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Autores principales:	Tatsuma Yao, Rie Suzuki, Natsuki Furuta, Yuka Suzuki, Kyoko Kabe, Mikiko Tokoro, Atsushi Sugawara, Akira Yajima, Tomohiro Nagasawa, Satoko Matoba, Kazuo Yamagata, Satoshi Sugimura
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2018
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/525032a9434c444f9740c9fa9e2e76e4
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spelling	oai:doaj.org-article:525032a9434c444f9740c9fa9e2e76e42021-12-02T11:40:15ZLive-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos10.1038/s41598-018-25698-w2045-2322https://doaj.org/article/525032a9434c444f9740c9fa9e2e76e42018-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-018-25698-whttps://doaj.org/toc/2045-2322Abstract Nuclear/chromosomal integrity is an important prerequisite for the assessment of embryo quality in artificial reproductive technology. However, lipid-rich dark cytoplasm in bovine embryos prevents its observation by visible light microscopy. We performed live-cell imaging using confocal laser microscopy that allowed long-term imaging of nuclear/chromosomal dynamics in bovine in vitro fertilised (IVF) embryos. We analysed the relationship between nuclear/chromosomal aberrations and in vitro embryonic development and morphological blastocyst quality. Three-dimensional live-cell imaging of 369 embryos injected with mRNA encoding histone H2B-mCherry and enhanced green fluorescent protein (EGFP)-α-tubulin was performed from single-cell to blastocyst stage for eight days; 17.9% reached the blastocyst stage. Abnormalities in the number of pronuclei (PN), chromosomal segregation, cytokinesis, and blastomere number at first cleavage were observed at frequencies of 48.0%, 30.6%, 8.1%, and 22.2%, respectively, and 13.0%, 6.2%, 3.3%, and 13.4%, respectively, for abnormal embryos developed into blastocysts. A multivariate analysis showed that abnormal chromosome segregation (ACS) and multiple PN correlated with delayed timing and abnormal blastomere number at first cleavage, respectively. In morphologically transferrable blastocysts, 30–40% of embryos underwent ACS and had abnormal PN. Live-cell imaging may be useful for analysing the association between nuclear/chromosomal dynamics and embryonic development in bovine embryos.Tatsuma YaoRie SuzukiNatsuki FurutaYuka SuzukiKyoko KabeMikiko TokoroAtsushi SugawaraAkira YajimaTomohiro NagasawaSatoko MatobaKazuo YamagataSatoshi SugimuraNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 8, Iss 1, Pp 1-9 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Tatsuma Yao Rie Suzuki Natsuki Furuta Yuka Suzuki Kyoko Kabe Mikiko Tokoro Atsushi Sugawara Akira Yajima Tomohiro Nagasawa Satoko Matoba Kazuo Yamagata Satoshi Sugimura Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
description	Abstract Nuclear/chromosomal integrity is an important prerequisite for the assessment of embryo quality in artificial reproductive technology. However, lipid-rich dark cytoplasm in bovine embryos prevents its observation by visible light microscopy. We performed live-cell imaging using confocal laser microscopy that allowed long-term imaging of nuclear/chromosomal dynamics in bovine in vitro fertilised (IVF) embryos. We analysed the relationship between nuclear/chromosomal aberrations and in vitro embryonic development and morphological blastocyst quality. Three-dimensional live-cell imaging of 369 embryos injected with mRNA encoding histone H2B-mCherry and enhanced green fluorescent protein (EGFP)-α-tubulin was performed from single-cell to blastocyst stage for eight days; 17.9% reached the blastocyst stage. Abnormalities in the number of pronuclei (PN), chromosomal segregation, cytokinesis, and blastomere number at first cleavage were observed at frequencies of 48.0%, 30.6%, 8.1%, and 22.2%, respectively, and 13.0%, 6.2%, 3.3%, and 13.4%, respectively, for abnormal embryos developed into blastocysts. A multivariate analysis showed that abnormal chromosome segregation (ACS) and multiple PN correlated with delayed timing and abnormal blastomere number at first cleavage, respectively. In morphologically transferrable blastocysts, 30–40% of embryos underwent ACS and had abnormal PN. Live-cell imaging may be useful for analysing the association between nuclear/chromosomal dynamics and embryonic development in bovine embryos.
format	article
author	Tatsuma Yao Rie Suzuki Natsuki Furuta Yuka Suzuki Kyoko Kabe Mikiko Tokoro Atsushi Sugawara Akira Yajima Tomohiro Nagasawa Satoko Matoba Kazuo Yamagata Satoshi Sugimura
author_facet	Tatsuma Yao Rie Suzuki Natsuki Furuta Yuka Suzuki Kyoko Kabe Mikiko Tokoro Atsushi Sugawara Akira Yajima Tomohiro Nagasawa Satoko Matoba Kazuo Yamagata Satoshi Sugimura
author_sort	Tatsuma Yao
title	Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
title_short	Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
title_full	Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
title_fullStr	Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
title_full_unstemmed	Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
title_sort	live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos
publisher	Nature Portfolio
publishDate	2018
url	https://doaj.org/article/525032a9434c444f9740c9fa9e2e76e4
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Live-cell imaging of nuclear–chromosomal dynamics in bovine in vitro fertilised embryos

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