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Nat. Methods, 08 January 2024 | https://doi.org/10.1038/s41592-023-02122-4 Real-time visualization of structural dynamics of synapses in live cells in vivo

Seungkyu Son ^1,7, Kenichiro Nagahama ^2,7, Jinsu Lee ^1,7, Kanghoon Jung ^2,3,7, Chuljung Kwak ², Jihoon Kim ¹, Young Woo Noh ^1,4, Eunjoon Kim^1,4, Sangkyu Lee ^5,*, Hyung-Bae Kwon ^2,*& Won Do Heo ^1,6,*

¹Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.
²Department of Neuroscience, Johns Hopkins School of Medicine, Baltimore, MD, USA.
³Allen Institute for Neural Dynamics, Seattle, WA, USA.
⁴Center for Synaptic Brain Dysfunctions, Institute for Basic Science, Daejeon, Republic of Korea.
⁵Center for Cognition and Sociality, Institute for Basic Science, Daejeon, Republic of Korea.
⁶KAIST Institute for the BioCentury, KAIST, Daejeon, Republic of Korea.
⁷These authors contributed equally: Seungkyu Son, Kenichiro Nagahama, Jinsu Lee, Kanghoon Jung.

^*Corresponding authors: correspondence to Sangkyu Lee, Hyung-Bae Kwon or Won Do Heo

Abstract

The structural plasticity of synapses is crucial for regulating brain functions. However, currently available methods for studying synapse organization based on split fluorescent proteins (FPs) have been limited in assessing synaptic dynamics in vivo due to the irreversible binding of split FPs. Here, we develop ‘SynapShot’, a method for visualizing the structural dynamics of intact synapses by combining dimerization-dependent FPs (ddFPs) with engineered synaptic adhesion molecules. SynapShot allows real-time monitoring of reversible and bidirectional changes of synaptic contacts under physiological stimulation. The application of green and red ddFPs in SynapShot enables simultaneous visualization of two distinct populations of synapses. Notably, the red-shifted SynapShot is highly compatible with blue light-based optogenetic techniques, allowing for visualization of synaptic dynamics while precisely controlling specific signaling pathways. Furthermore, we demonstrate that SynapShot enables real-time monitoring of structural changes in synaptic contacts in the mouse brain during both primitive and higher-order behaviors.

논문정보

형식Research article
게재일2024년 01월 (BRIC 등록일 2024-01-10)
연구진국내(교신)+국외 연구진
분야 생명과학 > 세포생물학

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