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

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. Summary: Microfluidics, live imaging and systems biology techniques are combined to develop a new approach for the functional analysis of sequence variants in the highly conserved Ras signaling pathway.

• Publication year

  2017

• Venue

  Disease Models & Mechanisms

• Publication date

  2017-07-01

• Fields of study

  Biology, Medicine, Engineering

• Identifiers
  DOI 10.1242/dmm.030163PMID 28495673PMCID 5536913
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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• In vivo severity ranking of Ras pathway mutations associated with developmental disorders

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• The Spatiotemporal Limits of Developmental Erk Signaling.

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  2016cited by this paper
• RASopathies: unraveling mechanisms with animal models

  2015cited by this paper
• RASopathies: unraveling mechanisms with animal models

  2015cited by this paper
• The RASopathies.

  2013cited by this paper
• Microfluidic trap array for massively parallel imaging of Drosophila embryos

  2013cited by this paper
• of Enzyme Kinetics in Cells ReviewERK as a Model for Systems Biology

  2013cited by this paper
• Kinetics of gene derepression by ERK signaling

  2013cited by this paper
• Torso RTK controls Capicua degradation by changing its subcellular localization

  2012influential reference
• The Capicua repressor – a general sensor of RTK signaling in development and disease

  2012cited by this paper
• MEK genomics in development and disease

  2012cited by this paper
• Global Tissue Revolutions in a Morphogenetic Movement Controlling Elongation

  2011cited by this paper
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  2011cited by this paper
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  2011cited by this paper
• Pulsed forces timed by a ratchet-like mechanism drive directed tissue movement during dorsal closure.

  2009cited by this paper
• In and out of Torso RTK signalling

  2003cited by this paper
• CIC, a member of a novel subfamily of the HMG-box superfamily, is transiently expressed in developing granule neurons.

  2002cited by this paper
• Relief of gene repression by torso RTK signaling: role of capicua in Drosophila terminal and dorsoventral patterning.

  2000cited by this paper
• Regulated Expression of nullo Is Required for the Formation of Distinct Apical and Basal Adherens Junctions in the Drosophila Blastoderm

  2000cited by this paper
• Multivariate Analysis

  1998cited by this paper
• Genetics of RAS signaling in C. elegans.

  1998cited by this paper
• MAP kinase in situ activation atlas during Drosophila embryogenesis.

  1997cited by this paper
• Reiterative use of the EGF receptor triggers differentiation of all cell types in the Drosophila eye.

  1996cited by this paper
• The ksr-7 Gene Encodes a Novel Protein Kinase Involved in Ras-Mediated Signaling in C. elegans

  1995cited by this paper
• A gain-of-function mutation in Drosophila MAP kinase activates multiple receptor tyrosine kinase signaling pathways.

  1994cited by this paper
• The activities of two Ets-related transcription factors required for Drosophila eye development are modulated by the Ras/MAPK pathway.

  1994cited by this paper
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  1993cited by this paper
• let-60, a gene that specifies cell fates during C. elegans vulval induction, encodes a ras protein.

  1990cited by this paper
• Localized surface activity of torso, a receptor tyrosine kinase, specifies terminal body pattern in Drosophila.

  1989cited by this paper
• Function of torso in determining the terminal anlagen of the Drosophila embryo

  1988cited by this paper
• Germline autonomy of maternal-effect mutations altering the embryonic body pattern of Drosophila.

  1986cited by this paper

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• Fly-on-a-Chip: Microfluidics for Drosophila melanogaster Studies.

  2019cites this paper
• Microfluidics as an Emerging Precision Tool in Developmental Biology.

  2019cites this paper
• A quantitative model of developmental RTK signaling.

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• Analyzing pathogenic MEK variants in zebrafish

  2017cites this paper
• How activating mutations affect MEK1 regulation and function

  2017cites this paper