Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation

Lindsey J. Macpherson; Emanuela E. Zaharieva; Patrick J. Kearney; Michael H. Alpert; Tzu Yang Lin; Zeynep Turan; Chi Hon Lee; Marco Gallio

doi:10.1038/ncomms10024

Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation

Lindsey J. Macpherson, Emanuela E. Zaharieva, Patrick J. Kearney, Michael H. Alpert, Tzu Yang Lin, Zeynep Turan, Chi Hon Lee, Marco Gallio^*

^*Corresponding author for this work

Neurobiology

Research output: Contribution to journal › Article › peer-review

144 Scopus citations

Abstract

Determining the pattern of activity of individual connections within a neural circuit could provide insights into the computational processes that underlie brain function. Here, we develop new strategies to label active synapses by trans-synaptic fluorescence complementation in Drosophila. First, we demonstrate that a synaptobrevin-GRASP chimera functions as a powerful activity-dependent marker for synapses in vivo. Next, we create cyan and yellow variants, achieving activity-dependent, multi-colour fluorescence reconstitution across synapses (X-RASP). Our system allows for the first time retrospective labelling of synapses (rather than whole neurons) based on their activity, in multiple colours, in the same animal. As individual synapses often act as computational units in the brain, our method will promote the design of experiments that are not possible using existing techniques. Moreover, our strategies are easily adaptable to circuit mapping in any genetic system.

Original language	English (US)
Article number	10024
Journal	Nature communications
Volume	6
DOIs	https://doi.org/10.1038/ncomms10024
State	Published - Dec 4 2015

Funding

This work was initially conceived and carried out in the laboratory of Charles Zuker, supported by NIH grants RC1NS069014 and R01NS076774. Work in the Gallio Lab was supported by NIH grant R01NS086859 to M.G. Additional support: the Intramural Research Programs of the NIH and the Eunice Kennedy Shriver National Institute of Child Health and Human Development (grant Z01-HD008776 to C.-H.L.). L.J.M. was a fellow of the Jane Coffin Childs Foundation. M.H.A. is supported by training grant T32HL007909. We thank Charles Zuker for experimental suggestions, discussions and support. We thank A. Kuang, M. Schmidt and B. Dick, HHMI EXROP students C. Washington and C. Gonzalez, summer intern M. Morgenstern for technical help, I. de la Rosa for fly maintenance and R. Barretto for insightful comments.

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

Access to Document

10.1038/ncomms10024

Cite this

@article{0020320015314b13b4afb72500db5055,

title = "Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation",

abstract = "Determining the pattern of activity of individual connections within a neural circuit could provide insights into the computational processes that underlie brain function. Here, we develop new strategies to label active synapses by trans-synaptic fluorescence complementation in Drosophila. First, we demonstrate that a synaptobrevin-GRASP chimera functions as a powerful activity-dependent marker for synapses in vivo. Next, we create cyan and yellow variants, achieving activity-dependent, multi-colour fluorescence reconstitution across synapses (X-RASP). Our system allows for the first time retrospective labelling of synapses (rather than whole neurons) based on their activity, in multiple colours, in the same animal. As individual synapses often act as computational units in the brain, our method will promote the design of experiments that are not possible using existing techniques. Moreover, our strategies are easily adaptable to circuit mapping in any genetic system.",

author = "Macpherson, {Lindsey J.} and Zaharieva, {Emanuela E.} and Kearney, {Patrick J.} and Alpert, {Michael H.} and Lin, {Tzu Yang} and Zeynep Turan and Lee, {Chi Hon} and Marco Gallio",

note = "Funding Information: This work was initially conceived and carried out in the laboratory of Charles Zuker, supported by NIH grants RC1NS069014 and R01NS076774. Work in the Gallio Lab was supported by NIH grant R01NS086859 to M.G. Additional support: the Intramural Research Programs of the NIH and the Eunice Kennedy Shriver National Institute of Child Health and Human Development (grant Z01-HD008776 to C.-H.L.). L.J.M. was a fellow of the Jane Coffin Childs Foundation. M.H.A. is supported by training grant T32HL007909. We thank Charles Zuker for experimental suggestions, discussions and support. We thank A. Kuang, M. Schmidt and B. Dick, HHMI EXROP students C. Washington and C. Gonzalez, summer intern M. Morgenstern for technical help, I. de la Rosa for fly maintenance and R. Barretto for insightful comments.",

year = "2015",

month = dec,

day = "4",

doi = "10.1038/ncomms10024",

language = "English (US)",

volume = "6",

journal = "Nature communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

}

TY - JOUR

T1 - Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation

AU - Macpherson, Lindsey J.

AU - Zaharieva, Emanuela E.

AU - Kearney, Patrick J.

AU - Alpert, Michael H.

AU - Lin, Tzu Yang

AU - Turan, Zeynep

AU - Lee, Chi Hon

AU - Gallio, Marco

N1 - Funding Information: This work was initially conceived and carried out in the laboratory of Charles Zuker, supported by NIH grants RC1NS069014 and R01NS076774. Work in the Gallio Lab was supported by NIH grant R01NS086859 to M.G. Additional support: the Intramural Research Programs of the NIH and the Eunice Kennedy Shriver National Institute of Child Health and Human Development (grant Z01-HD008776 to C.-H.L.). L.J.M. was a fellow of the Jane Coffin Childs Foundation. M.H.A. is supported by training grant T32HL007909. We thank Charles Zuker for experimental suggestions, discussions and support. We thank A. Kuang, M. Schmidt and B. Dick, HHMI EXROP students C. Washington and C. Gonzalez, summer intern M. Morgenstern for technical help, I. de la Rosa for fly maintenance and R. Barretto for insightful comments.

PY - 2015/12/4

Y1 - 2015/12/4

N2 - Determining the pattern of activity of individual connections within a neural circuit could provide insights into the computational processes that underlie brain function. Here, we develop new strategies to label active synapses by trans-synaptic fluorescence complementation in Drosophila. First, we demonstrate that a synaptobrevin-GRASP chimera functions as a powerful activity-dependent marker for synapses in vivo. Next, we create cyan and yellow variants, achieving activity-dependent, multi-colour fluorescence reconstitution across synapses (X-RASP). Our system allows for the first time retrospective labelling of synapses (rather than whole neurons) based on their activity, in multiple colours, in the same animal. As individual synapses often act as computational units in the brain, our method will promote the design of experiments that are not possible using existing techniques. Moreover, our strategies are easily adaptable to circuit mapping in any genetic system.

AB - Determining the pattern of activity of individual connections within a neural circuit could provide insights into the computational processes that underlie brain function. Here, we develop new strategies to label active synapses by trans-synaptic fluorescence complementation in Drosophila. First, we demonstrate that a synaptobrevin-GRASP chimera functions as a powerful activity-dependent marker for synapses in vivo. Next, we create cyan and yellow variants, achieving activity-dependent, multi-colour fluorescence reconstitution across synapses (X-RASP). Our system allows for the first time retrospective labelling of synapses (rather than whole neurons) based on their activity, in multiple colours, in the same animal. As individual synapses often act as computational units in the brain, our method will promote the design of experiments that are not possible using existing techniques. Moreover, our strategies are easily adaptable to circuit mapping in any genetic system.

UR - http://www.scopus.com/inward/record.url?scp=84949310074&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84949310074&partnerID=8YFLogxK

U2 - 10.1038/ncomms10024

DO - 10.1038/ncomms10024

M3 - Article

C2 - 26635273

AN - SCOPUS:84949310074

SN - 2041-1723

VL - 6

JO - Nature communications

JF - Nature communications

M1 - 10024

ER -

Dynamic labelling of neural connections in multiple colours by trans-synaptic fluorescence complementation

Abstract

Funding

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this