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

Yogesh Goyal, Thomas J. Levario, Henry H. Mattingly, Susan Holmes, Stanislav Y. Shvartsman, Hang Lu

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The Ras pathway patterns the poles of the Drosophila 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.

Original languageEnglish (US)
Pages (from-to)923-929
Number of pages7
JournalDMM Disease Models and Mechanisms
Volume10
Issue number7
DOIs
StatePublished - Jul 1 2017

All Science Journal Classification (ASJC) codes

  • Neuroscience (miscellaneous)
  • Medicine (miscellaneous)
  • Immunology and Microbiology (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)

Keywords

  • Capicua
  • Microfluidics
  • Mutations
  • Ras
  • SVD

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