Assessing heterogeneity among single embryos and single blastomeres using open microfluidic design
| Autor: | Lin He, Elisabet Rosàs-Canyelles, Alisha Geldert, Andrew J. Modzelewski, Amy E. Herr |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0303 health sciences
Messenger RNA Multidisciplinary Zygote Microfluidics Embryogenesis SciAdv r-articles Embryo Blastomere Biology Phenotype Cell biology 03 medical and health sciences Multicellular organism 0302 clinical medicine Engineering 030217 neurology & neurosurgery Research Articles 030304 developmental biology Research Article Developmental Biology |
| Zdroj: | Science Advances |
| ISSN: | 2375-2548 |
| Popis: | We report a microfluidic platform that maps target protein and mRNA expression with inter- and intraembryonic resolution. 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|