Multiplexed and high-throughput neuronal fluorescence imaging with diffusible probes
Name
s41467-019-12372-6.pdf
Description
Published version
Size
8.69 MB
Format
Adobe PDF
Checksum (MD5)
a66c56d774cbc07f00acb32c33ce8db7
Author(s)
Guo, Syuan-Ming
Veneziano, Remi
Gordonov, Simon
Li, Li
Danielson, Eric W
Perez de Arce, Karen
Park, Demian
Kulesa, Anthony Benjamin
Wamhoff, Eike-Christian
Blainey, Paul C.
Date Issued
September 2019
Publisher
Springer Science and Business Media LLC
Citation
Guo, Syuang-Ming et al. "Multiplexed and high-throughput neuronal fluorescence imaging with diffusible probes." Nature Communications 10, 4377 (September 2019): 4377 © 2019 The Author(s)
Version
Final published version
Abstract
Synapses contain hundreds of distinct proteins whose heterogeneous expression levels are determinants of synaptic plasticity and signal transmission relevant to a range of diseases. Here, we use diffusible nucleic acid imaging probes to profile neuronal synapses using multiplexed confocal and super-resolution microscopy. Confocal imaging is performed using high-affinity locked nucleic acid imaging probes that stably yet reversibly bind to oligonucleotides conjugated to antibodies and peptides. Super-resolution PAINT imaging of the same targets is performed using low-affinity DNA imaging probes to resolve nanometer-scale synaptic protein organization across nine distinct protein targets. Our approach enables the quantitative analysis of thousands of synapses in neuronal culture to identify putative synaptic sub-types and co-localization patterns from one dozen proteins. Application to characterize synaptic reorganization following neuronal activity blockade reveals coordinated upregulation of the post-synaptic proteins PSD-95, SHANK3 and Homer-1b/c, as well as increased correlation between synaptic markers in the active and synaptic vesicle zones.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Media Laboratory
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Terms of Use
Creative Commons Attribution 4.0 International license
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DOI of Published Version
https://doi.org/10.1038/s41467-019-12372-6
