Parallel imaging of embryos for quantitative analysis of genetic perturbations of the Ras pathway.

TitleParallel imaging of embryos for quantitative analysis of genetic perturbations of the Ras pathway.
Publication TypeJournal Article
Year of Publication2017
AuthorsGoyal, Y, Levario, TJ, Mattingly, HH, Holmes, S, Shvartsman, SY, Lu, H
JournalDis Model Mech
Volume10
Issue7
Pagination923-929
Date Published2017 07 01
ISSN1754-8411
KeywordsAnimals, Drosophila melanogaster, Drosophila Proteins, Embryo, Nonmammalian, HMGB Proteins, Image Processing, Computer-Assisted, Imaging, Three-Dimensional, Lab-On-A-Chip Devices, Mutation, ras Proteins, Repressor Proteins, Signal Transduction, Single-Cell Analysis, Time-Lapse Imaging
Abstract

<p>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.</p>

DOI10.1242/dmm.030163
Alternate JournalDis Model Mech
PubMed ID28495673
PubMed Central IDPMC5536913
Grant ListF31 AA023160 / AA / NIAAA NIH HHS / United States
R01 GM088333 / GM / NIGMS NIH HHS / United States
R01 GM107103 / GM / NIGMS NIH HHS / United States
R01 NS096581 / NS / NINDS NIH HHS / United States