RRC ID 54990
Author Farrar MJ, Kolkman KE, Fetcho JR.
Title Features of the structure, development, and activity of the zebrafish noradrenergic system explored in new CRISPR transgenic lines.
Journal J Comp Neurol
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 two-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 interhemispheric 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.
Volume 526(15)
Pages 2493-2508
Published 2018-10-15
DOI 10.1002/cne.24508
PMID 30070695
PMC PMC6196102
MeSH Aging Animals Animals, Genetically Modified Calcium Signaling / genetics Calcium Signaling / physiology Clustered Regularly Interspaced Short Palindromic Repeats* Larva / physiology Locus Coeruleus / cytology Locus Coeruleus / metabolism Locus Coeruleus / physiology Neurites / physiology Neurons / physiology Neurotransmitter Agents / metabolism Norepinephrine / physiology* Norepinephrine Plasma Membrane Transport Proteins / genetics Parasympathetic Nervous System / anatomy & histology Parasympathetic Nervous System / growth & development Parasympathetic Nervous System / physiology* Plasmids Rhombencephalon / anatomy & histology Rhombencephalon / cytology Rhombencephalon / physiology Zebrafish / physiology*
IF 3.4
Times Cited 4
Zebrafish Tg(vglut2a:loxP-DsRed-loxP-GFP) Tg(gad1b:GFP)