Stefan Sachs, Sebastian Reinhard, Janna Eilts, Markus Sauer, Christian Werner
Frontiers in Cellular Neuroscience, Vol. 18 · 2024
High-fidelity synaptic neurotransmission in the millisecond range is provided by a defined structural arrangement of synaptic proteins. At the presynapse multi-epitope scaffolding proteins are organized spatially at release sites to guarantee optimal binding of neurotransmitters at receptor clusters. The organization of pre- and postsynaptic proteins in trans-synaptic nanocolumns would thus intuitively support efficient information transfer at the synapse. Visualization of these protein-dense regions as well as the minute size of protein-packed synaptic clefts remains, however, challenging. To enable efficient labeling of these protein complexes, we developed post-gelation immunolabeling expansion microscopy combined with Airyscan super-resolution microscopy. Using \textasciitilde8-fold expanded samples, Airyscan enables multicolor fluorescence imaging with 20-30 nm spatial resolution. Post-immunolabeling of decrowded (expanded) samples provides increased labeling efficiency and allows the visualization of trans-synaptic nanocolumns. Our approach is ideally suited to investigate the pathological impact on nanocolumn arrangement e.g. in limbic encephalitis with autoantibodies targeting trans-synaptic leucine-rich glioma inactivated 1 protein (LGI1).
@article{sachs_visualizing_2024,
title = {Visualizing the trans-synaptic arrangement of synaptic proteins by expansion microscopy},
volume = {18},
issn = {1662-5102},
url = {https://www.frontiersin.org/articles/10.3389/fncel.2024.1328726},
doi = {10.3389/fncel.2024.1328726},
abstract = {High-fidelity synaptic neurotransmission in the millisecond range is provided by a defined structural arrangement of synaptic proteins. At the presynapse multi-epitope scaffolding proteins are organized spatially at release sites to guarantee optimal binding of neurotransmitters at receptor clusters. The organization of pre- and postsynaptic proteins in trans-synaptic nanocolumns would thus intuitively support efficient information transfer at the synapse. Visualization of these protein-dense regions as well as the minute size of protein-packed synaptic clefts remains, however, challenging. To enable efficient labeling of these protein complexes, we developed post-gelation immunolabeling expansion microscopy combined with Airyscan super-resolution microscopy. Using \textasciitilde8-fold expanded samples, Airyscan enables multicolor fluorescence imaging with 20-30 nm spatial resolution. Post-immunolabeling of decrowded (expanded) samples provides increased labeling efficiency and allows the visualization of trans-synaptic nanocolumns. Our approach is ideally suited to investigate the pathological impact on nanocolumn arrangement e.g. in limbic encephalitis with autoantibodies targeting trans-synaptic leucine-rich glioma inactivated 1 protein (LGI1).},
language = {English},
urldate = {2024-04-06},
journal = {Frontiers in Cellular Neuroscience},
author = {Sachs, Stefan and Reinhard, Sebastian and Eilts, Janna and Sauer, Markus and Werner, Christian},
year = {2024},
note = {Publisher: Frontiers},
keywords = {Airyscan, expansion microscopy, Nanocolumns, super-resolution microscopy, synapse, trans-synaptic},
}