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Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design
The process by which a zygote develops from a single cell into a multicellular organism is poorly understood. Advances are hindered by detection specificity and sensitivity limitations of single-cell protein tools and by challenges in integrating multimodal data. We introduce an open microfluidic to...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Association for the Advancement of Science
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7176412/ https://www.ncbi.nlm.nih.gov/pubmed/32494630 http://dx.doi.org/10.1126/sciadv.aay1751 |
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author | Rosàs-Canyelles, Elisabet Modzelewski, Andrew J. Geldert, Alisha He, Lin Herr, Amy E. |
author_facet | Rosàs-Canyelles, Elisabet Modzelewski, Andrew J. Geldert, Alisha He, Lin Herr, Amy E. |
author_sort | Rosàs-Canyelles, Elisabet |
collection | PubMed |
description | The process by which a zygote develops from a single cell into a multicellular organism is poorly understood. Advances are hindered by detection specificity and sensitivity limitations of single-cell protein tools and by challenges in integrating multimodal data. We introduce an open microfluidic tool expressly designed for same-cell phenotypic, protein, and mRNA profiling. We examine difficult-to-study—yet critically important—murine preimplantation embryo stages. In blastomeres dissociated from less well-studied two-cell embryos, we observe no significant GADD45a protein expression heterogeneity, apparent at the four-cell stage. In oocytes, we detect differences in full-length versus truncated DICER-1 mRNA and protein, which are insignificant by the two-cell stage. Single-embryo analyses reveal intraembryonic heterogeneity, differences between embryos of the same fertilization event and between donors, and reductions in the burden of animal sacrifice. Open microfluidic design integrates with existing workflows and opens new avenues for assessing the cellular-to-molecular heterogeneity inherent to preimplantation embryo development. |
format | Online Article Text |
id | pubmed-7176412 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | American Association for the Advancement of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-71764122020-06-02 Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design Rosàs-Canyelles, Elisabet Modzelewski, Andrew J. Geldert, Alisha He, Lin Herr, Amy E. Sci Adv Research Articles The process by which a zygote develops from a single cell into a multicellular organism is poorly understood. Advances are hindered by detection specificity and sensitivity limitations of single-cell protein tools and by challenges in integrating multimodal data. We introduce an open microfluidic tool expressly designed for same-cell phenotypic, protein, and mRNA profiling. We examine difficult-to-study—yet critically important—murine preimplantation embryo stages. In blastomeres dissociated from less well-studied two-cell embryos, we observe no significant GADD45a protein expression heterogeneity, apparent at the four-cell stage. In oocytes, we detect differences in full-length versus truncated DICER-1 mRNA and protein, which are insignificant by the two-cell stage. Single-embryo analyses reveal intraembryonic heterogeneity, differences between embryos of the same fertilization event and between donors, and reductions in the burden of animal sacrifice. Open microfluidic design integrates with existing workflows and opens new avenues for assessing the cellular-to-molecular heterogeneity inherent to preimplantation embryo development. American Association for the Advancement of Science 2020-04-22 /pmc/articles/PMC7176412/ /pubmed/32494630 http://dx.doi.org/10.1126/sciadv.aay1751 Text en Copyright © 2020 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC). http://creativecommons.org/licenses/by-nc/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial license (http://creativecommons.org/licenses/by-nc/4.0/) , which permits use, distribution, and reproduction in any medium, so long as the resultant use is not for commercial advantage and provided the original work is properly cited. |
spellingShingle | Research Articles Rosàs-Canyelles, Elisabet Modzelewski, Andrew J. Geldert, Alisha He, Lin Herr, Amy E. Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
title | Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
title_full | Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
title_fullStr | Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
title_full_unstemmed | Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
title_short | Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
title_sort | assessing heterogeneity among single embryos and single blastomeres using open microfluidic design |
topic | Research Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7176412/ https://www.ncbi.nlm.nih.gov/pubmed/32494630 http://dx.doi.org/10.1126/sciadv.aay1751 |
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