Fig. 3

Fig. 3 Pcp4a modulates representation of visual stimuli size in the tectum (A) Image showing part of the tectum of a 7 dpf elavl3:H2B-GCaMP6s fish. (B) Scheme of the strategy used to identify pcp4a+ neurons with in situ HCR after in vivo calcium imaging. (C) Confocal images showing localization in a 7 dpf elavl3:H2B-GCaMP6s larva of pcp4a’s mRNA and GCaMP6s signal acquired during in vivo Ca2+ imaging. The images were taken from the image stack obtained after aligning the in vivo and post-mortem stacks. (D) Graph depicting cumulative percentages of WMR angles of pcp4a+ PVNs in food-deprived and fed 7 dpf elavl3:H2B-GCaMP6s larvae. nfed = 6 fish, nnot-fed = 5 fish. (E) Bar graph showing average percentages of pcp4a+ PVNs in fed (n = 6 fish) and not-fed (n = 5 fish) larvae. (F and G) Graphs depicting cumulative percentages of WMR angles of pcp4a+ (F) or pcp4a− (G) PVNs in unfed 7 dpf elavl3:H2B-GCaMP6s; pcp4a+/+ and elavl3:H2B-GCaMP6s; pcp4amd78/md78 larvae. npcp4a+/+ = 6 fish, npcp4amd78/md78 = 6 fish. (H) Confocal image showing SINs in the tectum of a 7 dpf elavl3:H2B-GCaMP6s larva. The dashed white line demarcates the neuropil region containing SINs. (I) Bar graph showing average percentages of pcp4a+ SINs in fed (n = 6 fish) and not-fed (n = 5 fish) larvae. (J) Graph depicting cumulative percentages of WMR angles of SINs in food-deprived 7 dpf elavl3:H2B-GCaMP6s; pcp4a+/+ and elavl3:H2B-GCaMP6s; pcp4amd78/md78 larvae. npcp4a+/+ = 6 fish, npcp4amd78/md78 = 6 fish. (K) Graph depicting cumulative percentages of WMR angles of pcp4a+ PVNs in food-deprived 7 dpf elavl3:H2B-GCaMP6s; pcp4a+/+ and elavl3:H2B-GCaMP6s; pcp4amd78/md78 larvae, treated with the CaMKII inhibitor KN-93 or control solution. ncontrol_pcp4a+/+ = 5 fish, ncontrol_pcp4amd78/md78 = 4 fish, nKN-93_pcp4a+/+ = 6 fish, nKN-93_pcp4amd78/md78 = 6 fish. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001; n.s., not significant; two-tailed t test (E and I) or two-way ANOVA (D, F, G, J, and K). Data are presented as mean ± standard error of the mean. Scale bars, 50 μm in (A) and (H) and 10 μm in (C). In (A) and (H), A and P indicate anterior and posterior directions, respectively.