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

Cody Saraceno,

Vladimir A. Timoshevskiy, Jeramiah J. Smith

et al.

Journal of Experimental Zoology Part B Molecular and Developmental Evolution, Journal Year: 2023, Volume and Issue: 342(3), P. 260 - 270

Published: Oct. 30, 2023

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

Language: Английский

Culture time to optimize embryo cell-free DNA analysis for frozen-thawed blastocysts undergoing noninvasive preimplantation genetic testing for aneuploidy DOI
Goli Ardestani,

Maria Banti,

Carmen M. García-Pascual

et al.

Fertility and Sterility, Journal Year: 2024, Volume and Issue: 122(3), P. 465 - 473

Published: May 7, 2024

Language: Английский

Citations

3
Trophectoderm biopsy of blastocysts following IVF and embryo culture increases epigenetic dysregulation in a mouse model DOI
Eric A. Rhon‐Calderón,

Cassidy N. Hemphill,

Lisa A. Vrooman

et al.

Human Reproduction, Journal Year: 2023, Volume and Issue: 39(1), P. 154 - 176

Published: Nov. 22, 2023

Abstract STUDY QUESTION Does trophectoderm biopsy (TEBx) of blastocysts for preimplantation genetic testing in the clinic affect normal placental and embryo development offspring metabolic outcomes a mouse model? SUMMARY ANSWER TEBx impacts embryonic health during early development, with some alterations resolving others worsening later triggering changes adult offspring. WHAT IS KNOWN ALREADY Previous studies have not assessed epigenetic morphological either human populations or animal models. DESIGN, SIZE, DURATION We employed model to identify effects IVF. Three groups were assessed: naturally conceived (Naturals), IVF, IVF + TEBx, at two developmental timepoints: day (E)12.5 (n = 40/Naturals, n 36/IVF, 36/IVF TEBx) E18.5 42/Naturals, 30/IVF, 35/IVF TEBx). Additionally, mimic clinical practice, we fourth group: Vitrification (Vit) E12.5 29) that combines vitrification. To assess effect health, characterized 12-week-old cohort 24/Naturals, 25/IVF PARTICIPANTS/MATERIALS, SETTING, METHODS Our used CF-1 females as egg donors SJL/B6 males sperm donors. vitrification performed using standardized methods. Placenta morphology was evaluated by hematoxylin–eosin staining, situ hybridization Tpbpa junctional zone marker immunohistochemistry CD34 fetal endothelial cell markers. For molecular analysis placentas embryos, DNA methylation analyzed pyrosequencing, luminometric assay, chip array technology. Expression patterns ascertained RNA sequencing. Triglycerides, total cholesterol, high-, low-, very low-density lipoprotein, insulin, glucose determined commercially available kits. MAIN RESULTS AND THE ROLE OF CHANCE observed E12.5, had worse outcome terms differential gene expression whole embryos compared alone Naturals. These reflected blood vessel density. At E18.5, fetuses maintained exacerbated. With respect placentas, changes, although different Naturals, equivalent group, except density, which persisted. Of note is most differences sex specific. conclude has more detrimental mid-gestation tissues, tissues persisting stages alone, addition after results pronounced potentially effects: these are significantly Finally, 12-week offspring, regardless sex, showed higher glucose, triglycerides, lower high-density lipoprotein only having body weight findings additionally support need impact new procedures ART ensure healthy pregnancies outcomes. LARGE SCALE DATA Data reported this work been deposited NCBI Gene Omnibus under accession number GSE225318. LIMITATIONS, REASONS FOR CAUTION This study mimics many humans, where on possible. WIDER IMPLICATIONS FINDINGS highlights importance assaying their placenta FUNDING/COMPETING INTEREST(S) funded National Centers Translational Research Reproduction Infertility grant P50 HD068157-06A1 (M.S.B., C.C., M.M.), Ruth L. Kirschstein Service Award Individual Postdoctoral Fellowship F32 HD107914 (E.A.R.-C.) HD089623 (L.A.V.), Institutes Health Training program Cell Molecular Biology T32 GM007229 (C.N.H.). No conflict interest.

Language: Английский

Citations

6
Mechanism of chromosomal mosaicism in preimplantation embryos and its effect on embryo development DOI
Xue Zhang, Peng‐Sheng Zheng

Journal of Assisted Reproduction and Genetics, Journal Year: 2024, Volume and Issue: 41(5), P. 1127 - 1141

Published: Feb. 22, 2024

Language: Английский

Citations

2
Noninvasive time-lapse 3D subcellular analysis of embryo development for machine learning-enabled prediction of blastocyst formation DOI Creative Commons

C. Y. Gregory Lee,

Geon Kim,

Taeseop Shin

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 8, 2024

Abstract In developmental biology and in vitro fertilization (IVF), image-based assessment of embryos is pivotal. Traditional methods clinical IVF have been constrained to 2D morpho-kinetic profiling manual selection, hindered by the absence noninvasive techniques for quantitative 3D imaging over extended durations. Here, we overcome these limitations employing low-coherence holotomography monitor mouse preimplantation embryo development from 2-cell stage expanded blastocyst. This approach enables generation refractive index tomograms unlabeled embryos, facilitating observation subcellular dynamics. We investigated spatiotemporal profiles development, identifying key parameters that distinguish between with differing outcomes—specifically, Grade A successfully progressed blastocysts within 72 hours, C did not. Using machine learning, demonstrate can offer a noninvasive, framework predicting high potential.

Language: Английский

Citations

2
The first lineage determination in mammals DOI Creative Commons
Peter Pfeffer

Developmental Biology, Journal Year: 2024, Volume and Issue: 513, P. 12 - 30

Published: May 17, 2024

This review describes in detail the morphological, cytoskeletal and gene expression events leading to regulatory network bifurcation point of trophoblast inner cell mass cells a variety mammalian preimplantation embryos. The interrelated processes compaction polarity establishment are discussed terms how they affect YAP/WWTR activity location fate cells. Comparisons between mouse, human, cattle, pig rabbit embryos suggest conserved role for signalling induction eutherian animals though mechanisms for, timing of, activation differs among species. Downstream targets show further differences, with marker GATA3 being direct target all examined mammals, while CDX2-positive SOX2-negative regulation varies.

Language: Английский

Citations

2
Imaging actin organisation and dynamics in 3D DOI Creative Commons

Thomas Phillips,

Stefania Marcotti, Susan Cox

et al.

Journal of Cell Science, Journal Year: 2024, Volume and Issue: 137(2)

Published: Jan. 15, 2024

ABSTRACT The actin cytoskeleton plays a critical role in cell architecture and the control of fundamental processes including division, migration survival. dynamics organisation F-actin have been widely studied breadth types on classical two-dimensional (2D) surfaces. Recent advances optical microscopy enabled interrogation these cytoskeletal networks cells within three-dimensional (3D) scaffolds, tissues vivo. Emerging studies indicate that dimensionality experienced by has profound impact structure function cytoskeleton, with 3D environments exhibiting arrangements differ to 2D environments. However, addition third (and fourth, time) dimension leads challenges sample preparation, imaging analysis, necessitating additional considerations achieve required signal-to-noise ratio spatial temporal resolution. Here, we summarise current tools for context highlight examples importance this understanding biology opportunities domain.

Language: Английский

Citations

1
Cell Competition Eliminates Aneuploid Human Pluripotent Stem Cells DOI Creative Commons
Amanda Ya, Chenhui Deng, Kristina Godek

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: May 10, 2024

Human pluripotent stem cells (hPSCs) maintain diploid populations for generations despite a persistently high rate of mitotic errors that cause aneuploidy, or chromosome imbalances. Consequently, to genome stability, aneuploidy must inhibit hPSC proliferation, but the mechanisms are unknown. Here, we surprisingly find homogeneous aneuploid hPSCs proliferate unlike non-transformed somatic cells. Instead, in mosaic populations, cell non-autonomous competition between neighboring and eliminates less fit Aneuploid with lower Myc higher p53 levels relative neighbors outcompeted conversely gain selective advantage when abundance switches. Thus, although frequently missegregate chromosomes inherently tolerate Myc- p53-driven preserves their integrity. These findings have important implications use regenerative medicine how human embryos established prevalence during early development.

Language: Английский

Citations

1
Building the brain mosaic: an expanded view DOI
Sahibjot Sran, Amanda Ringland, Tracy A. Bedrosian

et al.

Trends in Genetics, Journal Year: 2024, Volume and Issue: 40(9), P. 747 - 756

Published: June 12, 2024

Language: Английский

Citations

1
Revisiting trophectoderm-inner cell mass lineage segregation in the mammalian preimplantation embryo DOI
Robin M. Skory

Human Reproduction, Journal Year: 2024, Volume and Issue: 39(9), P. 1889 - 1898

Published: June 26, 2024

In the first days of life, cells mammalian embryo segregate into two distinct lineages, trophectoderm and inner cell mass. Unlike nonmammalian species, development does not proceed from predetermined factors in oocyte. Rather, asymmetries arise de novo early incorporating cues position, contractility, polarity, cell-cell contacts. Molecular heterogeneities, including transcripts non-coding RNAs, have now been characterized as 2-cell stage. However, it's debated whether these heterogeneities bias toward one fate or other lineage identity arises stochastically at 16-cell This review summarizes what is known about blastomere our understanding allocation context historical models. Preimplantation reviewed coupled with changes morphology, transcription factor networks. The addition single-cell atlases human embryos has begun to reveal key differences between mouse, timing events core factors. Furthermore, recent generation blastoid models will provide valuable tools test understand determinants. Lastly, new techniques are reviewed, which may better synthesize existing knowledge emerging data sets reconcile regulative capacity unique embryo.

Language: Английский

Citations

1
The current clinical applications of preimplantation genetic testing (PGT): acknowledging the limitations of biology and technology DOI
Georgia Kakourou, Christalena Sofocleous, Thalia Mamas

et al.

Expert Review of Molecular Diagnostics, Journal Year: 2024, Volume and Issue: 24(9), P. 767 - 775

Published: Aug. 7, 2024

Preimplantation Genetic Testing (PGT) is a cutting-edge test used to detect genetic abnormalities in embryos fertilized through Medically Assisted Reproduction (MAR). PGT aims ensure that selected for transfer are free of specific conditions or chromosome abnormalities, thereby reducing chances unsuccessful MAR cycles, complicated pregnancies, and diseases future children.

Language: Английский

Citations

1