Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering

Neuron. 2020 Aug 5;107(3):454-469.e6. doi: 10.1016/j.neuron.2020.05.005. Epub 2020 Jun 22.


Neuroscience relies on techniques for imaging the structure and dynamics of neural circuits, but the cell bodies of individual neurons are often obscured by overlapping fluorescence from axons and dendrites in surrounding neuropil. Here, we describe two strategies for using the ribosome to restrict the expression of fluorescent proteins to the neuronal soma. We show first that a ribosome-tethered nanobody can be used to trap GFP in the cell body, thereby enabling direct visualization of previously undetectable GFP fluorescence. We then design a ribosome-tethered GCaMP for imaging calcium dynamics. We show that this reporter faithfully tracks somatic calcium dynamics in the mouse brain while eliminating cross-talk between neurons caused by contaminating neuropil. In worms, this reporter enables whole-brain imaging with faster kinetics and brighter fluorescence than commonly used nuclear GCaMPs. These two approaches provide a general way to enhance the specificity of imaging in neurobiology.

Keywords: C. elegans; GCaMP; calcium imaging; ribo-GCaMP; ribosomal tagging; soma-targeting; whole-brain imaging.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Brain / diagnostic imaging*
  • Brain / metabolism
  • Brain / pathology
  • Caenorhabditis elegans
  • Calcium / metabolism*
  • Calcium-Binding Proteins
  • Cell Body / metabolism
  • Cell Body / pathology*
  • Green Fluorescent Proteins
  • Mice
  • Neurons / metabolism
  • Neurons / pathology*
  • Neuropil
  • Optical Imaging / methods*
  • Ribosomal Protein L10 / metabolism
  • Ribosomes / metabolism*
  • Single-Domain Antibodies


  • Calcium-Binding Proteins
  • Single-Domain Antibodies
  • Green Fluorescent Proteins
  • Calcium