Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections

Nagatoki Kinoshita, Arthur J.Y. Huang, Thomas J. McHugh, Sachihiro C. Suzuki, Ichiro Masai, Il Hwan Kim, Scott H. Soderling, Atsushi Miyawaki, Tomomi Shimogori

Research output: Contribution to journalArticle

Abstract

Intercellular contacts are essential for precise organ morphogenesis, function, and maintenance; however, spatiotemporal information of cell-cell contacts or adhesions remains elusive in many systems. We developed a genetically encoded fluorescent indicator for intercellular contacts with optimized intercellular GFP reconstitution using glycosylphosphatidylinositol (GPI)anchor, GRAPHIC (GPI anchored reconstitution-activated proteins highlight intercellular connections), which can be used for an expanded number of cell types. We observed a robust GFP signal specifically at the interface between cultured cells, without disrupting natural cell contact. Application of GRAPHIC to the fish retina specifically delineated cone-bipolar connection sites. Moreover, we showed that GRAPHIC can be used in the mouse central nervous system to delineate synaptic sites in the thalamocortical circuit. Finally, we generated GRAPHIC color variants, enabling detection of multiple convergent contacts simultaneously in cell culture system. We demonstrated that GRAPHIC has high sensitivity and versatility, which will facilitate the analysis of the complex multicellular connections without previous limitations.

Original languageEnglish (US)
Pages (from-to)28-38
Number of pages11
JournaliScience
Volume15
DOIs
StatePublished - May 31 2019

Fingerprint

Glycosylphosphatidylinositols
Proteins
Morphogenesis
Retina
Cultured Cells
Fishes
Central Nervous System
Color
Cell Culture Techniques
Cell Count
Maintenance

All Science Journal Classification (ASJC) codes

  • General

Cite this

Kinoshita, N., Huang, A. J. Y., McHugh, T. J., Suzuki, S. C., Masai, I., Kim, I. H., ... Shimogori, T. (2019). Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections. iScience, 15, 28-38. https://doi.org/10.1016/j.isci.2019.04.013

Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections. / Kinoshita, Nagatoki; Huang, Arthur J.Y.; McHugh, Thomas J.; Suzuki, Sachihiro C.; Masai, Ichiro; Kim, Il Hwan; Soderling, Scott H.; Miyawaki, Atsushi; Shimogori, Tomomi.

In: iScience, Vol. 15, 31.05.2019, p. 28-38.

Research output: Contribution to journalArticle

Kinoshita, N, Huang, AJY, McHugh, TJ, Suzuki, SC, Masai, I, Kim, IH, Soderling, SH, Miyawaki, A & Shimogori, T 2019, 'Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections', iScience, vol. 15, pp. 28-38. https://doi.org/10.1016/j.isci.2019.04.013
Kinoshita, Nagatoki ; Huang, Arthur J.Y. ; McHugh, Thomas J. ; Suzuki, Sachihiro C. ; Masai, Ichiro ; Kim, Il Hwan ; Soderling, Scott H. ; Miyawaki, Atsushi ; Shimogori, Tomomi. / Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections. In: iScience. 2019 ; Vol. 15. pp. 28-38.
@article{9cc4c33bba5b40bfa682f353a22b97c0,
title = "Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections",
abstract = "Intercellular contacts are essential for precise organ morphogenesis, function, and maintenance; however, spatiotemporal information of cell-cell contacts or adhesions remains elusive in many systems. We developed a genetically encoded fluorescent indicator for intercellular contacts with optimized intercellular GFP reconstitution using glycosylphosphatidylinositol (GPI)anchor, GRAPHIC (GPI anchored reconstitution-activated proteins highlight intercellular connections), which can be used for an expanded number of cell types. We observed a robust GFP signal specifically at the interface between cultured cells, without disrupting natural cell contact. Application of GRAPHIC to the fish retina specifically delineated cone-bipolar connection sites. Moreover, we showed that GRAPHIC can be used in the mouse central nervous system to delineate synaptic sites in the thalamocortical circuit. Finally, we generated GRAPHIC color variants, enabling detection of multiple convergent contacts simultaneously in cell culture system. We demonstrated that GRAPHIC has high sensitivity and versatility, which will facilitate the analysis of the complex multicellular connections without previous limitations.",
author = "Nagatoki Kinoshita and Huang, {Arthur J.Y.} and McHugh, {Thomas J.} and Suzuki, {Sachihiro C.} and Ichiro Masai and Kim, {Il Hwan} and Soderling, {Scott H.} and Atsushi Miyawaki and Tomomi Shimogori",
year = "2019",
month = "5",
day = "31",
doi = "10.1016/j.isci.2019.04.013",
language = "English (US)",
volume = "15",
pages = "28--38",
journal = "iScience",
issn = "2589-0042",
publisher = "Elsevier Inc.",

}

TY - JOUR

T1 - Genetically Encoded Fluorescent Indicator GRAPHIC Delineates Intercellular Connections

AU - Kinoshita, Nagatoki

AU - Huang, Arthur J.Y.

AU - McHugh, Thomas J.

AU - Suzuki, Sachihiro C.

AU - Masai, Ichiro

AU - Kim, Il Hwan

AU - Soderling, Scott H.

AU - Miyawaki, Atsushi

AU - Shimogori, Tomomi

PY - 2019/5/31

Y1 - 2019/5/31

N2 - Intercellular contacts are essential for precise organ morphogenesis, function, and maintenance; however, spatiotemporal information of cell-cell contacts or adhesions remains elusive in many systems. We developed a genetically encoded fluorescent indicator for intercellular contacts with optimized intercellular GFP reconstitution using glycosylphosphatidylinositol (GPI)anchor, GRAPHIC (GPI anchored reconstitution-activated proteins highlight intercellular connections), which can be used for an expanded number of cell types. We observed a robust GFP signal specifically at the interface between cultured cells, without disrupting natural cell contact. Application of GRAPHIC to the fish retina specifically delineated cone-bipolar connection sites. Moreover, we showed that GRAPHIC can be used in the mouse central nervous system to delineate synaptic sites in the thalamocortical circuit. Finally, we generated GRAPHIC color variants, enabling detection of multiple convergent contacts simultaneously in cell culture system. We demonstrated that GRAPHIC has high sensitivity and versatility, which will facilitate the analysis of the complex multicellular connections without previous limitations.

AB - Intercellular contacts are essential for precise organ morphogenesis, function, and maintenance; however, spatiotemporal information of cell-cell contacts or adhesions remains elusive in many systems. We developed a genetically encoded fluorescent indicator for intercellular contacts with optimized intercellular GFP reconstitution using glycosylphosphatidylinositol (GPI)anchor, GRAPHIC (GPI anchored reconstitution-activated proteins highlight intercellular connections), which can be used for an expanded number of cell types. We observed a robust GFP signal specifically at the interface between cultured cells, without disrupting natural cell contact. Application of GRAPHIC to the fish retina specifically delineated cone-bipolar connection sites. Moreover, we showed that GRAPHIC can be used in the mouse central nervous system to delineate synaptic sites in the thalamocortical circuit. Finally, we generated GRAPHIC color variants, enabling detection of multiple convergent contacts simultaneously in cell culture system. We demonstrated that GRAPHIC has high sensitivity and versatility, which will facilitate the analysis of the complex multicellular connections without previous limitations.

UR - http://www.scopus.com/inward/record.url?scp=85066266141&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85066266141&partnerID=8YFLogxK

U2 - 10.1016/j.isci.2019.04.013

DO - 10.1016/j.isci.2019.04.013

M3 - Article

VL - 15

SP - 28

EP - 38

JO - iScience

JF - iScience

SN - 2589-0042

ER -