Soma-Targeted Imaging of Neural Circuits by Ribosome Tethering
| Autor: | David E. Taylor, Perry W.E. Spratt, Ling Bai, Aygul Subkhangulova, Yiming Chen, Seher Kosar, Rachel A Essner, Heeun Jang, Jennifer L. Garrison, Saul Kato, Zachary A. Knight, Evan H. Feinberg, David E. Leib, Kevin J. Bender, Mili Patel, Yen-Chu Lin, Tzu-Wei Kuo |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Neuropil Ribosomal Protein L10 Somatic cell Green Fluorescent Proteins Ribosome Green fluorescent protein Mice 03 medical and health sciences 0302 clinical medicine Calcium imaging medicine Biological neural network Animals Caenorhabditis elegans Neurons Chemistry General Neuroscience Calcium-Binding Proteins Optical Imaging Brain Single-Domain Antibodies 030104 developmental biology medicine.anatomical_structure nervous system Cell Body GCaMP Calcium Soma Ribosomes Neuroscience 030217 neurology & neurosurgery |
| Zdroj: | Neuron. 107:454-469.e6 |
| ISSN: | 0896-6273 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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