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ZFIN ID: ZDB-PUB-180803-5
Features of the structure, development and activity of the Zebrafish Noradrenergic System explored in new CRISPR transgenic lines
Farrar, M.J., Kolkman, K.E., Fetcho, J.R.
Date: 2018
Source: The Journal of comparative neurology   526(15): 2493-2508 (Journal)
Registered Authors: Fetcho, Joseph R.
Keywords: CRISPR/Cas9, RRID:AB_141372, RRID:AB_143165, RRID:AB_221570, RRID:AB_528490, developmental patterning, in vivo imaging, noradrenergic system, transgenic zebrafish
MeSH Terms: none
PubMed: 30070695 Full text @ J. Comp. Neurol.
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ABSTRACT
The noradrenergic (NA) system of vertebrates is implicated in learning, memory, arousal, and neuroinflammatory responses, but is difficult to access experimentally. Small and optically transparent, larval zebrafish offer the prospect of exploration of NA structure and function in an intact animal. We made multiple transgenic zebrafish lines using the CRISPR/Cas9 system to insert fluorescent reporters upstream of slc6a2, the Norepinephrine Transporter gene. These lines faithfully express reporters in NA cell populations, including the locus coeruleus (LC), which contains only about 14 total neurons. We used the lines in combination with 2-photon microscopy to explore the structure and projections of the NA system in the context of the columnar organization of cell types in the zebrafish hindbrain. We found robust alignment of NA projections with glutamatergic neurotransmitter stripes in some hindbrain segments, suggesting orderly relations to neuronal cell types early in life. We also quantified neurite density in the rostral spinal cord in individual larvae with as much as 100% difference in the number of LC neurons, and found no correlation between neuronal number in the LC and projection density in the rostral spinal cord. Finally, using light sheet microscopy, we performed bilateral calcium imaging of the entire LC. We found that large-amplitude calcium responses were evident in all LC neurons and showed bilateral synchrony, whereas small-amplitude events were more likely to show inter-hemispheric asynchrony, supporting the potential for targeted LC neuromodulation. Our observations and new transgenic lines set the stage for a deeper understanding of the NA system. This article is protected by copyright. All rights reserved.
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