- Title
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Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain
- Authors
- Shahar, O.D., Schuman, E.M.
- Source
- Full text @ Elife
Cell-type-specific labeling of newly synthesized proteins. |
High resolution imaging newly synthesized proteins in different neuronal populations. |
Fluorescently labeled nascent proteins in neurons across the zebrafish larva brain. |
CFP Ab staining and ANL labeling were performed on WT zebrafish larvae. Shown are confocal images of the similar regions of the same dimensions (X, Y, Z) of the confocal images shown in |
3 dpf larvae were incubated with 10 mM ANL for 24 hr in vivo, fixed (at 4 dpf) and clicked to a fluorescent alkyne tag to label nascent proteins in situ. Shown are single planes of CFP Ab staining and nascent protein labeling of the regions in the images in |
Click chemistry was performed to label nascent proteins. Antibody staining for CFP was performed to detect cells expressing CFP (and MetRSL270G) across the larvae. Individual neurons were segmented in 3D according to the CFP signal, before measuring nascent proteins (in the segmented ROIs). ( |
Zoom-in on a more challenging densely labeled ROI in the habenula. ( |
Representative images of cells exhibiting labeling of nascent proteins in neurites. Maximum intensity projections of 3–6 confocal planes (5.7–12 microns) are shown. Freely swimming larvae were incubated with ANL for 24 hr (same larvae as in |
Larvae were incubated with 10 mM ANL in their water bath for the indicated durations before fixation and click reaction. ( |
Seizure-induced neuron-specific protein synthesis. |
Seizure-induced neuron-specific protein noise levels in WT. |