![]() ![]() Subpanels 2–3: mean confocal signal in grin2B control and grin2B−/− larvae, for excitatory (Tg(vglut:DsRed) green) and inhibitory (Tg(dl圆a-1.4kbdlx5a/dl圆a:GFP) magenta) neurons. Subpanel 1: distribution of cell-rich (white) and neuropil-rich (magenta) zones (ZBB). C–E Coronal sections through parts of the subpallium, including the anterior cell-rich domain (C), anterior commissure (D), and preoptic region (E). Subpanels 2–4 from (C–E) were generated using data acquired from dlx:GFP larvae. In Panel B, C1, D1, and E1 were generated using the Zebrafish Brain Browser. Anterior (A), posterior (P), dorsal (D), ventral (V). Planes through which coronal sections were reconstructed for panels C–E are indicated. B Reconstructed sagittal section from the Zebrafish Brain Browser (ZBB) showing the distribution of glutamatergic (green) and GABAergic (red) neurons in the larval brain. (*p = 0.022, independent-samples t test). Values are shown as the percent increase in the signal grin2B−/− larvae compared to grin2B control. A Heat map of the ratio of size, transgenic expression of excitatory (vGlut) and inhibitory (dlx) marker, and calculated excitatory/inhibitory balance (E/I) for grin2B control (n = 13) and grin2B−/− (n = 9) larvae. Whole-brain imaging of zebrafish lacking GluN2B reveal a reduced inhibitory neuron signal in the subpallium. The statistical analysis was performed using the ordinary one-way ANOVA test. The mean diameter of the VA was used for quantification. (E) Schematic and data quantification of the width and length of the OFT. The statistical analysis was performed using the two-way ANOVA test (mixed-effects analysis). Due to the variation in the diameter at different segments of the vessels, AA1 and ORA were separated into AA1.1 and AA1.2 and ORA.1 and ORA.2, respectively. (D) Schematic and data quantification of the diameter of the AA1, ORA, and AA3. The gray area represents the range of symmetric distributions, in which the differences of the distance between X and Y is less than 15%. Each spot in the scatter diagram indicates one single fish. (C) Quantification of the branching positions of the AA3 on the left and right side of WT, chd7 sr5, and MZchd7 sr5 as X and Y coordinates. AA, aortic arch AA1, mandibular arch ORA, opercular artery AA3, first branchial arch AA4, second branchial arch HA, hypobranchial artery VA, ventral aorta OFT, outflow tract V, ventricle and A, atrium. If the AA3 branches off from the ORA instead of the VA, a negative value is used to quantify the length. X = the length on the left side of the fish, Y = the length on the right side of the fish. The distance between the branching points of the ORA and AA3 along the VA was measured. The enlarged/zoomed in schematic illustrated the abnormal AA3 branching observed in some of the chd7 mutants. (B) Schematic of ventral view of the zebrafish vasculature system at 102 hpf. 20x stacked confocal image, number of stacks: WT n = 25 (interval = 5 µm), chd7 sr5/+ n = 17 (interval = 7.5 µm), chd7 sr5 n = 20 (interval = 7.5 µm), and MZchd7 sr5 n = 21 (interval = 7.5 µm). (A) Ventral view of the kdrl:mTurquois fish at 102 hpf. ![]() Chd7 sr5 mutants show defects in arch artery patterning and ventral aorta length. ![]()
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