Parallel imaging of embryos for quantitative analysis of genetic perturbations of the Ras pathway. Author Yogesh Goyal, Thomas Levario, Henry Mattingly, Susan Holmes, Stanislav Shvartsman, Hang Lu Publication Year 2017 Type Journal Article Abstract The Ras pathway patterns the poles of the embryo by downregulating the levels and activity of a DNA-binding transcriptional repressor Capicua (Cic). We demonstrate that the spatiotemporal pattern of Cic during this signaling event can be harnessed for functional studies of mutations in the Ras pathway in human diseases. Our approach relies on a new microfluidic device that enables parallel imaging of Cic dynamics in dozens of live embryos. We found that although the pattern of Cic in early embryos is complex, it can be accurately approximated by a product of one spatial profile and one time-dependent amplitude. Analysis of these functions of space and time alone reveals the differential effects of mutations within the Ras pathway. Given the highly conserved nature of Ras-dependent control of Cic, our approach provides new opportunities for functional analysis of multiple sequence variants from developmental abnormalities and cancers. Keywords Repressor Proteins, Animals, Drosophila Proteins, Signal Transduction, Mutation, Drosophila melanogaster, Image Processing, Computer-Assisted, ras Proteins, Embryo, Nonmammalian, Single-Cell Analysis, Time-Lapse Imaging, HMGB Proteins, Lab-On-A-Chip Devices, Imaging, Three-Dimensional Journal Dis Model Mech Volume 10 Issue 7 Pages 923-929 Date Published 2017 Jul 01 ISSN Number 1754-8411 DOI 10.1242/dmm.030163 Alternate Journal Dis Model Mech PMCID PMC5536913 PMID 28495673 PubMedPubMed CentralGoogle ScholarBibTeXEndNote X3 XML