Journal: The Journal of Comparative Neurology

Article Title: Evidence for Auditory Stimulus‐Specific Adaptation But Not Deviance Detection in Larval Zebrafish Brains

doi: 10.1002/cne.70046

Figure Lengend Snippet: Calcium imaging of responses to auditory stimuli. (a) Experimental setup. ( b) Example single plane of a zebrafish larval brain expressing nuclear localized GCaMP6f under the control of the HuC promoter. (c) Visualization of all neurons segmented from a single larva colored by depth. (d) Stimulus train. ( e) Average Δ F / F traces over all neurons ( n = 186,675) from all fish ( n = 12) for each block of the experiment. Note: (a) A larva mounted in low melting point agarose is illuminated by two perpendicular sheets of light. Acoustic stimuli are delivered by a speaker affixed to the back wall. Calcium activity is captured with a water‐immersion objective. (b) Image was averaged over the entire recording after motion correction. (c) Black is most ventral, white most dorsal. Example neurons are highlighted in red, with their respective calcium activity traces over time represented below. (d) Experimental tones of 531 Hz (pink) and 1189 Hz (purple) each act as the standard and deviant stimulus in different blocks. Control tones of 355 Hz, 794 Hz, and 1778 Hz enable each frequency to be presented at the same probability in the many standards control blocks. (e) Grey boxes indicate the timings of the stimulus blocks. Stimulus times for the experimental tones when not standard are illustrated above each graph.

Article Snippet: We generated larvae from a stock of adult transgenic zebrafish ( Danio rerio , Tg(elavl3:H2B‐GCaMP6f), ZFIN identifier: ZDB‐ALT‐150916‐4, RRID: Addgene_67159) with targeted expression of the fluorescent calcium indicator GCaMP6f in the nuclei of neurons (Chen et al. ), maintained on a TLN background at a density of 10–15 fish per liter.

Techniques: Imaging, Expressing, Control, Blocking Assay, Activity Assay