Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact

Abstract The ability to identify specific cell–cell contact in the highly heterogeneous mammalian body is crucial to revealing precise control of the body plan and correct function. To visualize local connections, we previously developed a genetically encoded fluorescent indicator, GRAPHIC, which la...

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Autores principales:	Nagatoki Kinoshita, Arthur J. Y. Huang, Thomas J. McHugh, Atsushi Miyawaki, Tomomi Shimogori
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1b91d93aa6a14636aa4ac2e4b8455e83
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spelling	oai:doaj.org-article:1b91d93aa6a14636aa4ac2e4b8455e832021-12-02T19:04:25ZDiffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact10.1038/s41598-020-71474-02045-2322https://doaj.org/article/1b91d93aa6a14636aa4ac2e4b8455e832020-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-71474-0https://doaj.org/toc/2045-2322Abstract The ability to identify specific cell–cell contact in the highly heterogeneous mammalian body is crucial to revealing precise control of the body plan and correct function. To visualize local connections, we previously developed a genetically encoded fluorescent indicator, GRAPHIC, which labels cell–cell contacts by restricting the reconstituted green fluorescent protein (GFP) signal to the contact site. Here, we modify GRAPHIC to give the reconstituted GFP motility within the membrane, to detect cells that make contact with other specific cells. Removal of leucine zipper domains, located between the split GFP fragment and glycophosphatidylinositol anchor domain, allowed GFP reconstituted at the contact site to diffuse throughout the entire plasma membrane, revealing cell morphology. Further, depending on the structural spacers employed, the reconstituted GFP could be selectively targeted to N terminal (NT)- or C terminal (CT)-probe-expressing cells. Using these novel constructs, we demonstrated that we can specifically label NT-probe-expressing cells that made contact with CT-probe-expressing cells in an epithelial cell culture and in Xenopus 8-cell-stage blastomeres. Moreover, we showed that diffusible GRAPHIC (dGRAPHIC) can be used in neuronal circuits to trace neurons that make contact to reveal a connection map. Finally, application in the developing brain demonstrated that the dGRAPHIC signal remained on neurons that had transient contacts during circuit development to reveal the contact history. Altogether, dGRAPHIC is a unique probe that can visualize cells that made specific cell–cell contact.Nagatoki KinoshitaArthur J. Y. HuangThomas J. McHughAtsushi MiyawakiTomomi ShimogoriNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nagatoki Kinoshita Arthur J. Y. Huang Thomas J. McHugh Atsushi Miyawaki Tomomi Shimogori Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact
description	Abstract The ability to identify specific cell–cell contact in the highly heterogeneous mammalian body is crucial to revealing precise control of the body plan and correct function. To visualize local connections, we previously developed a genetically encoded fluorescent indicator, GRAPHIC, which labels cell–cell contacts by restricting the reconstituted green fluorescent protein (GFP) signal to the contact site. Here, we modify GRAPHIC to give the reconstituted GFP motility within the membrane, to detect cells that make contact with other specific cells. Removal of leucine zipper domains, located between the split GFP fragment and glycophosphatidylinositol anchor domain, allowed GFP reconstituted at the contact site to diffuse throughout the entire plasma membrane, revealing cell morphology. Further, depending on the structural spacers employed, the reconstituted GFP could be selectively targeted to N terminal (NT)- or C terminal (CT)-probe-expressing cells. Using these novel constructs, we demonstrated that we can specifically label NT-probe-expressing cells that made contact with CT-probe-expressing cells in an epithelial cell culture and in Xenopus 8-cell-stage blastomeres. Moreover, we showed that diffusible GRAPHIC (dGRAPHIC) can be used in neuronal circuits to trace neurons that make contact to reveal a connection map. Finally, application in the developing brain demonstrated that the dGRAPHIC signal remained on neurons that had transient contacts during circuit development to reveal the contact history. Altogether, dGRAPHIC is a unique probe that can visualize cells that made specific cell–cell contact.
format	article
author	Nagatoki Kinoshita Arthur J. Y. Huang Thomas J. McHugh Atsushi Miyawaki Tomomi Shimogori
author_facet	Nagatoki Kinoshita Arthur J. Y. Huang Thomas J. McHugh Atsushi Miyawaki Tomomi Shimogori
author_sort	Nagatoki Kinoshita
title	Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact
title_short	Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact
title_full	Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact
title_fullStr	Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact
title_full_unstemmed	Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact
title_sort	diffusible graphic to visualize morphology of cells after specific cell–cell contact
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/1b91d93aa6a14636aa4ac2e4b8455e83
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Diffusible GRAPHIC to visualize morphology of cells after specific cell–cell contact

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