Additional file 1 of The gut metabolite indole-3-propionic acid activates ERK1 to restore social function and hippocampal inhibitory synaptic transmission in a 16p11.2 microdeletion mouse model
Creators: name: Jiang, Jian
name: Wang, Dilong
name: Jiang, Youheng
name: Yang, Xiuyan
name: Sun, Runfeng
name: Chang, Jinlong
name: Zhu, Wenhui
name: Yao, Peijia
name: Song, Kun
name: Chang, Shuwen
name: Wang, Hong
name: Zhou, Lei
name: Zhang, Xue-Song
name: Li, Huiliang
name: Li, Ningning
Publisher: name: figshare
Year: 2025
Subjects: subject: Biochemistry
subject: Medicine
subject: Genetics
subject: FOS: Biological sciences
subject: Neuroscience
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Cancer
subject: Mental Health

description: Additional file 1: Figure S1. 16p11.2 $${}^{+/-}$$ + / -  mice showed gut metabolites disturbance in feces. Changes in the relative abundance of differential metabolites. A Indole-3-propionic acid (IPA). B Adenosine. C Isobutyric acid. D 15-keto Prostaglandin E1 (15-keto-PGE1). E sn-Glycerol 3-phosphoethanolamine (DOPE). F 3-Phenylpropanoic acid. G Ketoisocaproic acid. H Perseitol. I D-Mannitol. J N-Acetyl-L-glutamate. K Scytalone. L N-Acetyl-L-Histidine. M 5-hydroperoxy-6,8,11,14-eicosatetraenoicacid (5(S)-HpETE). N Erucic acid. O 3-Hydroxycapric acid. P N-Oleoylethanolamine. Q Linoleoyl ethanolamide. R Vanillin. S Deoxyadenosine. T Uridine. (WT: n = 7 mice; 16p11.2: n = 9 mice. Student’s t test). Data are presented as mean ± SEM. *p &lt; 0.05 and **p &lt; 0.01. Detailed statistical information is presented in Additional file 2: Table S1. Figure S2. 16p11.2 $${}^{+/-}$$ + / -  mice displayed hyperactivity but no significant anxiety. A, B 16p11.2 $${}^{+/-}$$ + / -  mice showed hyperactivity in the open field test (OFT). C, D 16p11.2 $${}^{+/-}$$ + / -  mice showed no significant anxiety-like behavior in the elevated plus-maze (EPM) test (WT: n = 11 mice; 16p11.2 $${}^{+/-}$$ + / -  : n = 10 mice. Student’s t test). Data are presented as mean ± SEM. *p &lt; 0.05 and n.s.: not significant. Detailed statistical information is presented in Additional file 2: Table S1. Figure S3. 16p11.2 $${}^{+/-}$$ + / -  mice exhibited normal activation level in the mPFC and CA1, CA3 of hippocampus. A The red puncta indicate c-Fos+ neurons in the CA1 of hippocampus. Scale bar: 50μm. B The density of c-Fos+ neurons were not changed significantly in CA1 region of 16p11.2 $${}^{+/-}$$ + / -  mice hippocampus (WT: n = 5 mice; 16p11.2 $${}^{+/-}$$ + / -  : n = 4 mice. Student’s t test). C The red puncta indicate c-Fos+ neurons in the CA3 of hippocampus. D The number of c-Fos+ neurons in CA3 of hippocampal region had no distinct difference between the WT and 16p11.2 $${}^{+/-}$$ + / -  groups (WT: n = 5 mice; 16p11.2 $${}^{+/-}$$ + / - : n = 4 mice. Student’s t test). E The red puncta indicate c-Fos+ neurons in mPFC. F No significant changes of the number of c-Fos+ neurons in mPFC (WT: n = 4 mice; 16p11.2 $${}^{+/-}$$ + / - : n = 3 mice. Student’s t test). Data are presented as mean ± SEM. n.s.: not significant. Detailed statistical information is presented in Additional file 2: Table S1. Figure S4. The 16p11.2 $${}^{+/-}$$ + / -  mice did not show significant changes in either sEPSC frequency or amplitude, but the decay time and rise time of sIPSC were increased significantly. A Representative sEPSCs traces from granule cells in hippocampus of mice. Scale bars: 5 s, 10 pA. B Cumulative distribution of sEPSCs amplitudes. C. Cumulative distribution of sEPSCs frequencies (WT: n = 653 events from 9 cells of 5 mice; 16p11.2 $${}^{+/-}$$ + / - : n = 281 events from 7 cells of 6 mice. Kolmogorov-Smirnov test). Data are presented as mean ± SEM. Detailed statistical information is presented in Additional file 2: Table S1. Figure S5. The effect of IPA on social interaction in diverse behavioral tests. A IPA did not affect the preference of mice for stranger mouse (S1) or empty cage (E) in the sociability phase of TCT (WT+Vehicle: n = 20 mice; WT+IPA: n = 20 mice; 16p11.2 $${}^{+/-}$$ + / - +Vehicle: n = 18 mice; 16p11.2 $${}^{+/-}$$ + / - +IPA: n = 18 mice. Two-way ANOVA). B There was no significant difference in preference between two identical objects (Obj1, Obj2) in the habituation phase of NOR test (WT+Vehicle: n = 20 mice; WT+IPA: n = 20 mice; 16p11.2 $${}^{+/-}$$ + / - +Vehicle: n = 18 mice; 16p11.2 $${}^{+/-}$$ + / - +IPA: n = 18 mice. Two-way ANOVA). C Schematic diagram of USVs behavioral tests in male-female social interaction. D The total number of USVs in adult male mice during the first phase (female present) of the experiment, with or without IPA treatment. E The total number of USVs in adult male mice during Phase 2 (female removed). F The total number of USVs in adult male mice during Phase 3 (female returned) (WT+Vehicle: n = 10 mice; WT+IPA: n = 10 mice; 16p11.2 $${}^{+/-}$$ + / - +Vehicle: n = 8 mice; 16p11.2 $${}^{+/-}$$ + / - +IPA: n = 10 mice). Data are presented as mean ± SEM. *p &lt; 0.05, ****p &lt; 0.0001 and n.s.: not significant. Detailed statistical information is presented in Additional file 2: Table S1. Figure S6. Neurobiotin-labeling experiment showed that dendritic branching or the number of dendritic spines of granule cells in DG region was not affected by mouse genotypes or IPA administration. A, B Representative confocal images of dendritic branching (A) and quantitative analysis (B) (WT+Vehicle: n = 5 cells from 3 mice; WT+IPA: n = 12 cells from 4 mice; 16p11.2 $${}^{+/-}$$ + / - +Vehicle: n = 14 cells from 4 mice; 16p11.2 $${}^{+/-}$$ + / - +IPA: n = 7 cells from 3 mice). Scale bar: 50μm. C, D Representative confocal images of dendritic spines (C) and statistical results (D) showed no significant change in the density of dendritic spines in 16p11.2 $${}^{+/-}$$ + / -  mice, and IPA had no effect on the density of dendritic spines (WT+Vehicle: n = 10 cells from 5 mice; WT+IPA: n = 11 cells from 4 mice; 16p11.2 $${}^{+/-}$$ + / - +Vehicle: n = 14 cells from 4 mice; 16p11.2 $${}^{+/-}$$ + / - +IPA: n = 8 cells from 3 mice.). Scale bar: 10μm. Data are presented as mean ± SEM. n.s.: not significant. Detailed statistical information is presented in Additional file 2: Table S1. Figure S7. Golgi staining showed that the density of dendritic spines of granule cells in DG region was not affected by mouse genotypes and IPA administration. A Representative images of dendritic spines. Scale bar: 10μm. B There was no significant change in the density of dendritic spines in 16p11.2 $${}^{+/-}$$ + / -  mice, and IPA did not affect the density of dendritic spines (n = 8 slices from 4 mice per group). Data are presented as mean ± SEM. n.s.: not significant. Detailed statistical information is presented in Additional file 2: Table S1.