Morphological characteristics of shank3b ^−/− larvae and adult zebrafish. a–b Abnormal morphological changes in shank3b ^−/− and shank3b ^+/− larvae at ~ 1 dpf, including severe developmental delay, eye melanin reduction (blue arrow), and tail bending (red arrow) (+/+, N = 60; +/−, N = 50; −/−, N = 50). c–d Normal morphology and body length of shank3b ^+/+ , shank3b ^+/− , and shank3b ^−/− larvae at 3 dpf (c) and adults (6 mpf, male) (d) (N = 20 for each genotype). e–f Significantly enlarged brain size (e) but normal brain weight (f) in adult male shank3b ^−/− (6 mpf) compared to WT zebrafish (N = 30 for each genotype). *p < 0.05

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.

shank3b ^−/− adult zebrafish displayed reduced and repetitive locomotion activity in the open-field test. a Schematic diagram of the open-field test and thigmotaxis test of adult male zebrafish. In the analysis of thigmotaxis test, the area of the peripheral zone is equal to the center zone (dotted line). b–c shank3b^−/− zebrafish at 3.5 mpf showed significantly reduced velocity in the total 60 min period (b) and velocity per larva (c) in the open field (N = 13 for each group). d Representative traces of individual shank3b ^+/+ or shank3 ^−/− zebrafish in the thigmotaxis test. e Ratio for the time spent and distance traveled (periphery divided by the total zone) over 30 min in adult male zebrafish (3.5 mpf). N = 13 for each group. f–g Representative trace of different types of stereotyped behaviors of shank3b ^−/− adult male zebrafish (3.5 mpf). shank3b^−/− zebrafish had a significantly higher proportion of figure “8” and big circling movements than shank3b ^+/+ . N = 13 for each group. Data are shown as mean ± SEM; **p < 0.01, ****p < 0.0001

shank3b ^−/− zebrafish displayed social interaction defect. a–b Schematic of shoaling test (a) and significantly increased inter-fish distance of adult male shank3b^−/− zebrafish (3.5 mpf) (b). N = 18 for each group. c–f Schematic of social preference test of adult male zebrafish (3.5 mpf) (c). Heat map (d) shows that shank3b ^+/+ zebrafish displayed significant higher frequency near a group of zebrafish than shank3b ^−/− zebrafish. Time ratio (e) and distance ratio (f) in the conspecific sector were significantly reduced in shank3b ^−/− zebrafish compared to shank3b ^+/+ zebrafish. N = 16 for each group. g–h Schematic of kin recognition and preference test of adult male zebrafish (3.5 mpf) (g) and significantly reduced ratio of kin zone entering in shank3b ^−/− zebrafish compared to shank3 ^+/+ zebrafish (h). N = 10 for each group. Data are presented as mean ± SEM; ****p < 0.0001

shank3b deficiency altered the neurodevelopment in larvae. a–c Reduced RFP staining in shank3b ^−/− larvae compared to shank3b^+/+ larvae (1 dpf, 2 dpf, and 3 dpf) using Huc: RFP transgene line zebrafish. The difference is the most prominent at 1 dpf. Scale bar, 100 μm. a’–c’ RT-qPCR results of RFP expressions from (a–c) larvae. N = 8 for each group. Data are presented as mean ± SEM; *p < 0.05, ****p < 0.0001

ZFIN is incorporating published figure images and captions as part of an ongoing project. Figures from some publications have not yet been curated, or are not available for display because of copyright restrictions.