Ultrabright chemical labeling enables rapid neural connectivity profiling in large tissue samples

Area of Science:

• Neuroscience
• Molecular Biology
• Biotechnology

Background:

• Mapping neuronal connections is crucial for understanding brain function.
• Existing methods face challenges in speed, specificity, and scale for whole-brain analysis.

Purpose of the Study:

• To introduce LINCS, a novel technology for efficient and sparse labeling of individual neurons.
• To enable large-scale, high-resolution mapping of neural circuits in the mouse nervous system.

Main Methods:

• Developed LINCS using engineered biotin ligase for in vivo biotinylation and monovalent streptavidin staining.
• Integrated LINCS with tissue clearing and light-sheet microscopy for whole-mount imaging.
• Employed AAV and Cas9-mediated Cre knockout for stable, sparse neuronal labeling.

Main Results:

• Achieved rapid, ultrabright, and photostable labeling of specific cell types throughout the mouse brain and body.
• Enabled profiling of long-range neuronal projections in both central and peripheral nervous systems.
• Facilitated precise morphological reconstruction of individual neurons at scale.

Conclusions:

• LINCS significantly advances the capability for large-scale neural connectivity mapping.
• This technology lowers barriers and accelerates the study of mammalian neural circuits.
• LINCS provides a powerful toolkit for anatomical and functional dissection of complex neural networks.