Title | Parallel imaging of embryos for quantitative analysis of genetic perturbations of the Ras pathway. |
Publication Type | Journal Article |
Year of Publication | 2017 |
Authors | Goyal, Y, Levario, TJ, Mattingly, HH, Holmes, S, Shvartsman, SY, Lu, H |
Journal | Dis Model Mech |
Volume | 10 |
Issue | 7 |
Pagination | 923-929 |
Date Published | 2017 Jul 01 |
ISSN | 1754-8411 |
Keywords | Animals, 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> |
DOI | 10.1242/dmm.030163 |
Alternate Journal | Dis Model Mech |
PubMed ID | 28495673 |
PubMed Central ID | PMC5536913 |
Grant List | F31 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 |