Monitoring cell-cell contacts in vivo in transgenic animals
Abstract
We used a synthetic genetic system based on ligand-induced intramembrane proteolysis to monitor cell-cell contacts in animals. Upon ligand-receptor interaction in sites of cell-cell contact, the transmembrane domain of an engineered receptor is cleaved by intramembrane proteolysis and releases a protein fragment that regulates transcription in the interacting partners. We demonstrate that the system can be used to regulate gene expression between interacting cells both in vitro and in vivo, in transgenic Drosophila. We show that the system allows for detection of interactions between neurons and glia in the Drosophila nervous system. In addition, we observed that when the ligand is expressed in subsets of neurons with a restricted localization in the brain it leads to activation of transcription in a selected set of glial cells that interact with those neurons. This system will be useful to monitor cell-cell interactions in animals, and can be used to genetically manipulate interacting cells.
Additional Information
© 2016 The Company of Biologists Ltd. Received July 19, 2016. Accepted September 13, 2016. Published online September 22, 2016. Michael Elowitz provided the CHO-UAS-H2Bmcit and CHO-rat Delta cell lines. James Kochenderfer (National Cancer Institute, Bethesda, MD, USA) supplied the 1D3-28z.1-3 and the MSGV-CD19 plasmids. Marc Freeman provided the repo-LexA::GAD and alrm-LexA::GAD drivers. Wilm Stork, Lukas Neukomm, Marc Freeman and Elizabeth Hong provided advice on the Drosophila nervous system anatomy and genetics. Daniel Lee helped edit the manuscript. Imaging from Zeiss 710 was performed in the Biological Imaging Facility, with the support of the Caltech Beckman Institute and the Arnold and Mabel Beckman Foundation. Author contributions: CV conceived the project. CV and THH designed experiments. THH performed all experiments with cell culture and transgenic Drosophila. TV performed western blot analysis. THH and CL wrote the manuscript. This work was funded by the National Institutes of Health [1R21NS08485 to C.L.]. Deposited in PMC for release after 12 months. No competing interests.Attached Files
Published - 4073.full.pdf
Supplemental Material - 21/dev.142406.DC1/DEV142406supp.pdf
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Additional details
- PMCID
- PMC5117150
- Eprint ID
- 70604
- Resolver ID
- CaltechAUTHORS:20160927-101143198
- 1R21NS08485
- NIH
- Caltech Beckman Institute
- Arnold and Mabel Beckman Foundation
- Created
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2016-09-28Created from EPrint's datestamp field
- Updated
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2022-04-08Created from EPrint's last_modified field