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Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering


Authors: Yiming Chen, Heeun Jang, Perry W.E. Spratt, Seher Kosar, David E. Taylor, Rachel A. Essner, Ling Bai, David E. Leib, Tzu-Wei Kuo, Yen-Chu Lin, Mili Patel, Aygul Subkhangulova, Saul Kato, Evan H. Feinberg, Kevin J. Bender, Zachary A. Knight, Jennifer L. Garrison
Publication: Neuron
Date: June 22, 2020
Link to article: https://www.cell.com/neuron/fulltext/S0896-6273(20)30351-2?dgcid=raven_jbs_aip_…


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.

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