Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway
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 th...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
The Company of Biologists
2017-07-01
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| Series: | Disease Models & Mechanisms |
| Subjects: | |
| Online Access: | http://dmm.biologists.org/content/10/7/923 |
| _version_ | 1811234544518430720 |
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| author | Yogesh Goyal Thomas J. Levario Henry H. Mattingly Susan Holmes Stanislav Y. Shvartsman Hang Lu |
| author_facet | Yogesh Goyal Thomas J. Levario Henry H. Mattingly Susan Holmes Stanislav Y. Shvartsman Hang Lu |
| author_sort | Yogesh Goyal |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-12T11:39:17Z |
| format | Article |
| id | doaj.art-002b0135007542a4908c5398b66c98f0 |
| institution | Directory Open Access Journal |
| issn | 1754-8403 1754-8411 |
| language | English |
| last_indexed | 2024-04-12T11:39:17Z |
| publishDate | 2017-07-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Disease Models & Mechanisms |
| spelling | doaj.art-002b0135007542a4908c5398b66c98f02022-12-22T03:34:44ZengThe Company of BiologistsDisease Models & Mechanisms1754-84031754-84112017-07-0110792392910.1242/dmm.030163030163Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathwayYogesh Goyal0Thomas J. Levario1Henry H. Mattingly2Susan Holmes3Stanislav Y. Shvartsman4Hang Lu5 Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA Department of Statistics, Stanford University, Stanford, CA 94305, USA Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ 08544, USA School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA 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.http://dmm.biologists.org/content/10/7/923MicrofluidicsRasMutationsCapicuaSVD |
| spellingShingle | Yogesh Goyal Thomas J. Levario Henry H. Mattingly Susan Holmes Stanislav Y. Shvartsman Hang Lu Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway Disease Models & Mechanisms Microfluidics Ras Mutations Capicua SVD |
| title | Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway |
| title_full | Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway |
| title_fullStr | Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway |
| title_full_unstemmed | Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway |
| title_short | Parallel imaging of Drosophila embryos for quantitative analysis of genetic perturbations of the Ras pathway |
| title_sort | parallel imaging of drosophila embryos for quantitative analysis of genetic perturbations of the ras pathway |
| topic | Microfluidics Ras Mutations Capicua SVD |
| url | http://dmm.biologists.org/content/10/7/923 |
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