Human embryo live imaging reveals nuclear DNA shedding during blastocyst expansion and biopsy

Domingo-Muelas, Ana; Skory, Robin M.; Moverley, Adam A.; Ardestani, Goli; Pomp, Oz; Rubio, Carmen; Tetlak, Piotr; Hernandez, Blake; Rhon‐Calderón, Eric A.; Navarro-Sánchez, Luis; García-Pascual, Carmen M.; Bissière, Stéphanie; Bartolomei, Marisa S.; Sakkas, Denny; Simón, Carlos; Plachta, Nicolas

doi:10.1016/j.cell.2023.06.003

Cited by 16 publications

(13 citation statements)

References 82 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Domingo et al reported that offsprings were obtained from mouse embryos transplanted after live imaging. 11 However, they did not state whether the result was obtained from embryos observed from the two-pronuclear stage, and toxicity to cells might be stronger in early stages, such as the two-pronuclear stage. In this study, the three groups showed no significant difference in the time from nuclear envelope breakdown (NEBD) to the two-cell stage, and there was no significant difference in the proportion of embryos that reached the four-cell stage after live imaging.…”

Section: Discussionmentioning

confidence: 99%

“…Domingo et al. examined the localization of microfilaments by using fixed samples, mRNA injection, and SPY‐555 actin, and reported that the use of these three methods resulted in comparable findings 11 …”

Section: Discussionmentioning

confidence: 99%

“…Domingo et al examined the localization of microfilaments by using fixed samples, mRNA injection, and SPY-555 actin, and reported that the use of these three methods resulted in comparable findings. 11 We combined these reagents to observe mouse embryos to investigate the complex and dynamic interactions between the chromatin, tubulin, and actin cytoskeleton from the late fertilization period to the first cleavage (i.e., from the two-pronuclear to the two-cell stage). To our knowledge, compared with the previous studies utilizing these unique probes, our study reports a clearer and more dynamic observation of early embryogenesis, showing three elements simultaneously.…”

Section: Discussionmentioning

confidence: 99%

“…For observation, the drops were covered with Light Mineral Oil (FUJIFILM Irvine Scientific Inc., Saitama, Japan). Regarding DNA, while a previous study used SPY650‐DNA, 11 we used a NucleoSeeing (excitation wavelength 488–520 nm) to avoid interference with the excitation wavelength of 700 nm for the microtubule SiR‐700‐tubulin.…”

Section: Methodsmentioning

confidence: 99%

“…Several low‐toxicity and cell‐permeable reagents, for live cell imaging, targeting the DNA and cytoskeletal proteins have recently been developed 9,10 . Moreover, the use of these reagents to visualize DNA and microfilaments during the developmental process involving the transition from mouse two‐cell stage embryo to blastocyst‐stage embryo, and in the development of human blastocyst‐stage embryos, was reported in 2023 11 …”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

An approach for live imaging of first cleavage in mouse embryos using fluorescent chemical probes for DNA, microtubules, and microfilaments

Okabe,

Shirasawa,

Ono

et al. 2023

Reprod Medicine & Biology

View full text Add to dashboard Cite

PurposeDynamic morphological changes in the chromosome and cytoskeleton occur in mammals and humans during early embryonic development, and abnormalities such as embryonic chromosomal aneuploidy occur when development does not proceed normally. Visualization of the intracellular organelles and cytoskeleton allows elucidation of the development of early mammalian embryos. The behavior of the DNA and cytoskeleton in early mammalian embryos has conventionally been observed by injecting target molecule mRNAs, incorporating a fluorescent substance‐expressing gene, into embryos. In this study, we visualized the chronological behavior of male and female chromosome condensation in mouse embryos, beginning in the two‐pronuclear zygote, through the first division to the two‐cell stage, using fluorescent chemical probes to visualize the behavior of DNA, microtubules, and microfilaments.MethodMouse two‐pronuclear stage embryo were immersed in medium containing fluorescent chemical probes to visualize DNA, microtubules, and microfilaments. Observation was performed with a confocal microscope.ResultsThis method allowed us to observe how chromosome segregation errors in first somatic cell divisions in mouse embryos and enabled dynamic analysis of a phenomenon called lagging chromosomes.ConclusionsBy applying this method, we can observe any stage of embryonic development, which may provide new insights into embryonic development in other mammals.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

An approach for live imaging of first cleavage in mouse embryos using fluorescent chemical probes for DNA, microtubules, and microfilaments

Okabe,

Shirasawa,

Ono

et al. 2023

Reprod Medicine & Biology

View full text Add to dashboard Cite

show abstract

Functional analyses of the polycomb‐group genes in sea lamprey embryos undergoing programmed DNA loss

Saraceno,

Timoshevskiy,

Smith

2023

J Exp Zool Pt B

View full text Add to dashboard Cite

During early development, sea lamprey embryos undergo programmatic elimination of DNA from somatic progenitor cells in a process termed programmed genome rearrangement (PGR). Eliminated DNA eventually becomes condensed into micronuclei, which are then physically degraded and permanently lost from the cell. Previous studies indicated that many of the genes eliminated during PGR have mammalian homologs that are bound by polycomb repressive complex (PRC) in embryonic stem cells. To test whether PRC components play a role in the faithful elimination of germline‐specific sequences, we used a combination of CRISPR/Cas9 and lightsheet microscopy to investigate the impact of gene knockouts on early development and the progression through stages of DNA elimination. Analysis of knockout embryos for the core PRC2 subunits EZH, SUZ12, and EED show that disruption of all three genes results in an increase in micronucleus number, altered distribution of micronuclei within embryos, and an increase in micronucleus volume in mutant embryos. While the upstream events of DNA elimination are not strongly impacted by loss of PRC2 components, this study suggests that PRC2 plays a role in the later stages of elimination related to micronucleus condensation and degradation. These findings also suggest that other genes/epigenetic pathways may work in parallel during DNA elimination to mediate chromatin structure, accessibility, and the ultimate loss of germline‐specific DNA.

show abstract

Mechanism of chromosomal mosaicism in preimplantation embryos and its effect on embryo development

Zhang,

Zheng

2024

J Assist Reprod Genet

View full text Add to dashboard Cite

Human embryo live imaging reveals nuclear DNA shedding during blastocyst expansion and biopsy

Cited by 16 publications

References 82 publications

An approach for live imaging of first cleavage in mouse embryos using fluorescent chemical probes for DNA, microtubules, and microfilaments

An approach for live imaging of first cleavage in mouse embryos using fluorescent chemical probes for DNA, microtubules, and microfilaments

Functional analyses of the polycomb‐group genes in sea lamprey embryos undergoing programmed DNA loss

Mechanism of chromosomal mosaicism in preimplantation embryos and its effect on embryo development

Contact Info

Product

Resources

About