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Reproduction, fertility, and development2019; 31(12); 1840-1850; doi: 10.1071/RD19254

Assessing equine embryo developmental competency by time-lapse image analysis.

Abstract: The timing of early mitotic events during preimplantation embryo development is important for subsequent embryogenesis in many mammalian species, including mouse and human, but, to date, no study has closely examined mitotic timing in equine embryos from oocytes obtained by ovum pick-up. Here, cumulus-oocyte complexes were collected by transvaginal follicular aspiration, matured invitro and fertilised via intracytoplasmic sperm injection. Each fertilised oocyte was cultured up to the blastocyst stage and monitored by time-lapse imaging for the measurement of cell cycle intervals and identification of morphological criteria indicative of developmental potential. Of the 56 fertilised oocytes, 35 initiated mitosis and 11 progressed to the blastocyst stage. Analysis of the first three mitotic divisions in embryos that formed blastocysts determined that typical blastocyst timing (median±IQR) is 30.0±17.5min, 8.8±1.7h and 0.6±1.4h respectively. Frequent cellular fragmentation, multipolar divisions and blastomere exclusion suggested that equine embryos likely contend with a high incidence of chromosomal missegregation. Indeed, chromosome-containing micronuclei and multinuclei with extensive DNA damage were observed throughout preimplantation embryogenesis. This indicates that time-lapse image analysis may be used as a non-invasive method to assess equine embryo quality in future studies.
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Publication Date: 2019-11-25 PubMed ID: 31759400PubMed Central: PMC7170182DOI: 10.1071/RD19254Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research in question pertains to a detailed scrutiny of the developmental process of equine embryos, procured from oocytes through ovum pick-up. The central focus is on the timing of early mitotic events. The study ascertains timing standards for equine blastocysts and identifies chromosomal anomalies. The findings underscore how time-lapse image analysis can serve as an effective non-invasive method to evaluate the development competency of equine embryos.

Investigation Methodology

The research employed the technique of transvaginal follicular aspiration to source cumulus-oocyte complexes, which were matured in vitro and fertilised through intracytoplasmic sperm injection. The researchers used time-lapse imaging for the observation of the fertilised oocytes from the point of fertilization up to the blastocyst stage.

  • The cell cycle intervals were measured, and morphological criteria were identified that could hint at the developmental potential.
  • 56 fertilised oocytes were monitored, of which 35 initiated mitosis and 11 progressed to the blastocyst stage.

Key Findings

The primary observations from the study focussed on the first three mitotic divisions in embryos that reached the blastocyst stage.

  • The timing for a typical blastocyst was determined as 30.0±17.5 minutes, 8.8±1.7 hours, and 0.6±1.4 hours for each of the three phases respectively.
  • Findings also indicated a higher incidence of chromosomal missegregation in equine embryos, as evident by frequent cellular fragmentation, multipolar divisions, and blastomere exclusion.
  • The researchers observed chromosome-containing micronuclei and multinuclei associated with substantial DNA damage throughout the embryo’s preimplantation development.

Implications of the Research

This research concludes that time-lapse image analysis can serve as a non-invasive method to assess the quality of equine embryos. The study’s insights on the timing of mitotic events and potential chromosomal anomalies can direct future research on equine embryo competencies and aid the development of new therapeutic and developmental strategies.

Cite This Article

APA
Brooks KE, Daughtry BL, Metcalf E, Masterson K, Battaglia D, Gao L, Park B, Chavez SL. (2019). Assessing equine embryo developmental competency by time-lapse image analysis. Reprod Fertil Dev, 31(12), 1840-1850. https://doi.org/10.1071/RD19254

Publication

Reproduction, fertility, and development
ISSN: 1448-5990
NlmUniqueID: 8907465
Country: Australia
Language: English
Volume: 31
Issue: 12
Pages: 1840-1850

Researcher Affiliations

Brooks, Kelsey E
  • Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Beaverton, OR 97006, USA.
Daughtry, Brittany L
  • Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Beaverton, OR 97006, USA; and Department of Cell, Developmental and Cancer Biology, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA.
Metcalf, Elizabeth
  • Department of Obstetrics and Gynecology, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA.
Masterson, Keith
  • Department of Obstetrics and Gynecology, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA.
Battaglia, David
  • Department of Obstetrics and Gynecology, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA.
Gao, Lina
  • Bioinformatics and Biostatistics Core, Oregon National Primate Research Center, Beaverton, OR 97006, USA.
Park, Byung
  • Bioinformatics and Biostatistics Core, Oregon National Primate Research Center, Beaverton, OR 97006, USA.
Chavez, Shawn L
  • Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Beaverton, OR 97006, USA; and Department of Obstetrics and Gynecology, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA; and Department of Physiology and Pharmacology, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA; and Department of Biomedical Engineering, Oregon Health and Science University School of Medicine, Portland, OR 97239, USA; and Corresponding author. Email: chavesh@ohsu.edu.

MeSH Terms

  • Animals
  • Blastocyst / cytology
  • Blastocyst / ultrastructure
  • Blastomeres / cytology
  • Blastomeres / ultrastructure
  • Cells, Cultured
  • Cytokinesis / physiology
  • Embryo Culture Techniques / veterinary
  • Embryo, Mammalian
  • Embryonic Development / physiology
  • Female
  • Horses / embryology
  • Male
  • Microscopy / methods
  • Microscopy / veterinary
  • Quality Control
  • Sperm Injections, Intracytoplasmic / methods
  • Sperm Injections, Intracytoplasmic / veterinary
  • Time-Lapse Imaging / methods
  • Time-Lapse Imaging / veterinary

Grant Funding

  • R01 HD086073 / NICHD NIH HHS

Conflict of Interest Statement

Conflicts of interest. The authors declare no conflicts of interest.

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