Rosàs-Canyelles Elisabet, Modzelewski Andrew J, Geldert Alisha, He Lin, Herr Amy E
Department of Bioengineering, University of California Berkeley, Berkeley, CA 94720, USA.
The University of California Berkeley and University of California San Francisco Graduate Program in Bioengineering, Berkeley, CA 94720, USA.
Sci Adv. 2020 Apr 22;6(17):eaay1751. doi: 10.1126/sciadv.aay1751. eCollection 2020 Apr.
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.
从单细胞受精卵发育成多细胞生物的过程仍鲜为人知。单细胞蛋白质工具的检测特异性和灵敏度限制以及整合多模态数据的挑战阻碍了这方面的进展。我们推出了一种专门设计用于单细胞表型、蛋白质和mRNA分析的开放式微流控工具。我们研究了难以研究但至关重要的小鼠植入前胚胎阶段。在从研究较少的二细胞胚胎解离的卵裂球中，我们未观察到在四细胞阶段明显的GADD45a蛋白表达异质性。在卵母细胞中，我们检测到全长与截短的DICER-1 mRNA和蛋白质的差异，到二细胞阶段这些差异不显著。单胚胎分析揭示了胚胎内的异质性、同一受精事件的胚胎之间以及供体之间的差异，并且减少了动物牺牲的负担。开放式微流控设计与现有工作流程相结合，为评估植入前胚胎发育固有的细胞到分子异质性开辟了新途径。
