Fig. 5: L. plantarum and NaB modulated the expression of Cxcl16 in hippocampal astrocytes to affect partial defects in social behavior in ASD mice.

(A) Schematic of viral infection and optical fiber implantation. (B-C) Inject AAV-GFaABC1D-mCherry-5’miR-30a-shRNA1(Cxcl16)-3’miR-30a-WPREs into hippocampal astrocytes to reduce CXCL16 expression, and then compare social behavior of both groups. (B) Left: Statistical analysis of the time that mice interacted with the object and the stranger mouse1. n = 14–17 mice/group. Right: Representative tracks of the mice moved in the three-chamber. (C) Left: Statistical analysis of the time that mice interacted with the mouse1 and the mouse2. n = 14–17 mice/group. Right: Representative tracks of the mice moved in the three-chamber. (D-E) Inject rAAV-GFaABC1D-Cxcl16-P2A-mCherry-WPRE-SV40pA to upregulate Cxcl16 expression in the hippocampal astrocytes of MIA offspring, and then compare social behavior between groups. (D) Left: Statistical analysis of the time that mice interacted with the object and the stranger mouse1. n = 16–17 mice/group. Right: Representative tracks of the mice moved in the three-chamber. (E) Left: Statistical analysis of the time that mice interacted with familiar and novel mice. n = 16–17 mice/group. Right: Representative tracks of the mice moved in the three-chamber. (F-I) Inject rAAV-hSyn-GCaMp6s-WPRE-hGHpA to reveal Ca2+ signaling between neuronal activity. (F) Top: The heatmap illustrates the Ca2+ signals of the PIC offsprings when interacting with the object in the sociability stage. Bottom: The peri-event plot of the mean Ca2+ signals corresponding to the heatmaps (F top). n = 8 mice/group. (G) The same as F but for the stranger mouse1 in the sociability stage. n = 8 mice/group. (H) The same as (G) but in the social novelty stage. n = 8 mice/group. (I) The same as (H) but for the stranger mouse2. n = 8 mice/group. (J-M) The same as (F-I) but for PIC+ Cxcl16 experimental mice. n = 8 mice/group. (N-Q) The same as (F-I) but for wide-type (WT) experimental mice. n = 8 mice/group. Data were presented as mean ± SEM, and statistical significance was determined by Sidak’s multiple comparisons test (B-E), significant differences were classified as follows: ns p > 0.05, *p < 0.05, ***p < 0.001, ****p < 0.0001.