Fig. 3

Ogt deficient astrocytes impair hippocampal neurons and mice cognition. a Representative images of neurons with Golgi staining in CA1 region of Ctrl and cKO mice. Scale bar, 10 μm. b Representative images of Golgi-stained dendritic spines of second-order segment in the CA1 region of Ctrl and cKO mice. Scale bar, 1 μm. c–e Sholl analysis showed the overall decrease in the number of dendritic intersections per radius (c) and the number of dendrites per cell (d) and total length of dendrites (e) in the Golgi-stained neurons from the CA1 region of cKO mice compared with Ctrl mice. n = 15 neurons from 3 mice per group were analyzed. Values represent mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001; two-way ANOVA analysis followed by Sidak’s multiple-comparison test for c, F_{(1, 1400)} = 153.7; unpaired Student’s t-test for d, e. f Quantification results show the decreased dendritic spine density of cKO mice compared to that of Ctrl mice. Spine density was calculated by dividing the number of spines with the length of dendrite. n = 24 neurons from 3 mice per group were analyzed. Values represent mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001; unpaired Student’s t-test. g The escape latency during the training period of Ctrl and cKO adult mice. cKO mice required longer time to reach the platform starting from the first day of the 4-day training period. Ctrl/cKO, n = 10 mice. Values represent mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001; two-way ANOVA analysis followed by Sidak’s multiple-comparison test, F_{(3, 72)} = 23.06. h–j During the probe test, cKO mice required longer time to reach the platform (h), decreased time in target quadrant (i), and decreased numbers of crossing the platform (j). Ctrl mice, n = 10; cKO mice, n = 10. Values represent mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001; unpaired Student’s t-test for h, j and two-way ANOVA analysis followed by Sidak’s multiple-comparison test for i, F_{(3, 72)} = 30.62. k The percentage of spontaneous alternations in cKO mice was lower than that in Ctrl mice during the Y maze spontaneous alternation task test. Ctrl, n = 8 mice; cKO, n = 8 mice. Values represent mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001; unpaired Student’s t-test. l Representative heatmap shows the distribution of exploring time of Ctrl and cKO mice during the testing trial in Y maze spatial novelty preference test. m–o cKO mice showed the decreased percentage of exploring time (m), the number of entries (n) and distance (o) in the novel arm compared to those in Ctrl mice during the Y maze spatial novelty preference test. n = 8 mice for Ctrl/cKO group. Values represent mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; two-way ANOVA analysis followed by Sidak’s multiple-comparison test, F_{(2, 42)} = 67.41 for m, F_{(2, 42)} = 15.92 for n, F_{(2, 42)} = 10.84 for o. p In the passive avoidance task test, cKO mice displayed shorter latency to enter the dark compartment than that of Ctrl mice during the retention test while Ctrl and cKO mice showed no difference of the latency during the training session. n = 8 mice for Ctrl/cKO group. Values represent mean ± SEM; *p < 0.05; **p < 0.01; ***p < 0.001; two-way ANOVA analysis followed by Sidak’s multiple-comparison test, F_{(1, 28)} = 456.2