期刊
NEURON
卷 107, 期 3, 页码 454-+出版社
CELL PRESS
DOI: 10.1016/j.neuron.2020.05.005
关键词
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资金
- Nikon Imaging Center at the University of California, San Francisco (UCSF) [S10OD017993]
- Caenorhabditis Genetics Center (CGC) [P40OD010440]
- Howard Hughes Medical Institute
- NIH Institutional National Research Service Award [T32 AG000266]
- New York Stem Cell Foundation
- American Diabetes Association
- NIH [S10OD017993, DP2DK109533, R01DK106399, R01NS094781, R35GM119828, DP2MH119426, R01 NS109060, R01DA035913, R01MH112729, R21MH112117]
- American Federation for Aging Research
- Glenn Foundation for Medical Research
- E. Matilda Ziegler Foundation for the Blind
- Sandler Foundation
- Esther A. and Joseph Klingenstein Fund
- Brain and Behavior Research Foundation
- Whitehall Foundation
- Simons Foundation
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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