Multiplexed and high-throughput neuronal fluorescence imaging with diffusible probes.

Nat Commun
Authors
Keywords
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.

Year of Publication
2019
Journal
Nat Commun
Volume
10
Issue
1
Pages
4377
Date Published
2019 09 26
ISSN
2041-1723
DOI
10.1038/s41467-019-12372-6
PubMed ID
31558769
PubMed Central ID
PMC6763432
Links
Grant list
R24 MH106075 / MH / NIMH NIH HHS / United States
U01 MH106011 / MH / NIMH NIH HHS / United States
RM1 HG008525 / HG / NHGRI NIH HHS / United States
T32 GM087237 / GM / NIGMS NIH HHS / United States
P30 ES002109 / ES / NIEHS NIH HHS / United States
R01 NS087950 / NS / NINDS NIH HHS / United States
R01 MH112694 / MH / NIMH NIH HHS / United States