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

Scientific Reports
Tatsuma YaoSatoshi Sugimura

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 b...Continue Reading

References

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Aug 16, 2005·Genesis : the Journal of Genetics and Development·Kazuo YamagataAtsuo Ogura
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May 30, 2017·The Journal of Reproduction and Development·Satoshi SugimuraKei Imai

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Citations

Mar 11, 2021·Theriogenology·Ryosuke SuzukiSatoshi Sugimura
Jan 26, 2021·The Journal of Reproduction and Development·Yasumitsu MasudaRyo Nishimura
May 1, 2021·Journal of Developmental Biology·Claudia SpringerKilian Simmet
May 14, 2021·Frontiers in Veterinary Science·Yasumitsu MasudaRyo Nishimura
May 28, 2021·Biochemical and Biophysical Research Communications·Hiroki NagaiSatoshi Sugimura
Jun 1, 2021·Biology of Reproduction·Coline MahéMarie Saint-Dizier

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Methods Mentioned

BETA
confocal microscopy
imaging
light microscopy

Software Mentioned

R
ordinal
Nippon package
Fiji
ImageJ
stats
Volocity
pROC

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