Fig. 8: LpEV treatment improved behavioral deficits of Oxtr heterozygous mice (Oxtr+/−). | Experimental & Molecular Medicine

Fig. 8: LpEV treatment improved behavioral deficits of Oxtr heterozygous mice (Oxtr+/−).

From: Lactobacillus paracasei-derived extracellular vesicles reverse molecular and behavioral deficits in mouse models of autism spectrum disorder

Fig. 8

a, Experimental design. Oxtr+/− mice were orally administered LpEV (2.27 mg/kg/day, or 50 μg per 22 g body weight per day) for 3 weeks, and this treatment continued during behavioral testing. Behavioral tests were conducted sequentially: social interaction, social preference and repetitive tests. b–e, Social behaviors. Time spent exploring a social target compared with an empty cage (b and d) and time spent exploring a novel stranger versus a familiar one (c and e) in the three-chamber social behavior test. Data shown for male (b and c) and female (d and e) Oxtr+/− mice. Male: n = 6 animals per group. Female: WT, 5 animals; Drd2-KO, 5 animals; Drd2-KO + LpEV, 3 animals. Male (b and c). Sociability (b). WT versus Oxtr+/−. Two-way ANOVA, Fisher’s LSD post-hoc test. Genotype, F(1,20) = 23.23, P = 0.0001; social target, F(1,20) = 79.82, P < 0.0001; genotype × social target, F(1,20) = 22.32, P = 0.0001. Oxtr+/− versus Oxtr+/− + LpEV. Two-way ANOVA, Fisher’s LSD post-hoc test. LpEV, F(1,20) = 10.36, P = 0.0032; social target, F(1,20) = 88.54, P < 0.0001; LpEV × social target, F(1,20) = 16.33, P = 0.0006. Social preference (c). WT versus Oxtr+/−. Two-way ANOVA, Fisher’s LSD post-hoc test. Genotype, F(1,20) = 12.16, P = 0.0023; social target, F(1,20) = 15.66, P = 0.0008; genotype × social target, F(1,20) = 11.05, P = 0.0034. Oxtr+/− versus Oxtr+/− + LpEV. Two-way ANOVA, Fisher’s LSD post-hoc test. LpEV, F(1,20) = 4.487, P = 0.0469; social target, F(1,20) = 15.52, P = 0.0008; LpEV × social target, F(1,20) = 10.94, P = 0.0035. Female (d and e). Sociability (d). WT versus Oxtr+/−. Two-way ANOVA, Fisher’s LSD post-hoc test. Genotype, F(1,16) = 2.857, P = 0.1104; social target, F(1,16) = 27.65, P < 0.0001; genotype × social target, F(1,16) = 12.85, P = 0.0025. Oxtr+/− versus Oxtr+/− + LpEV. Two-way ANOVA, Fisher’s LSD post-hoc test. LpEV, F(1,12) = 2.089, P = 0.0704; social target, F(1,12) = 13.79, P = 0.0030; LpEV × social target, F(1,12) = 3.944, P = 0.0704. Social preference (e). WT versus Oxtr+/−. Two-way ANOVA, Fisher’s LSD post-hoc test. Genotype, F(1,16) = 6.901, P = 0.0183; social target, F(1,16) = 16.09, P = 0.0010; genotype × social target, F(1,16) = 5.007, P = 0.0398. Oxtr+/− versus Oxtr+/− + LpEV. Two-way ANOVA, Fisher’s LSD post-hoc test. LpEV, F(1,12) = 0.6644, P = 0.4309; social target, F(1,12) = 11.58, P = 0.0052; LpEV × social target, F(1,12) = 4.622, P = 0.0527. f–k, Repetitive behaviors. Time spent by male (f–h) and female (i–k) Oxtr+/− mice on grooming (f and i), rearing (g and j) and digging (h and k). Male: WT, 5 animals; Drd2-KO, 4 animals; Drd2-KO + LpEV5, 5 animals; Drd2-KO + LpEV15, 5 animals. Female: n = 5 animals per group. Male (f–h). WT versus Oxtr-KO (−/−) versus Oxtr-KO (+/−) versus Oxtr-KO (+/−) + LpEV. One-way ANOVA, Tukey’s post-hoc test. Grooming, F(3,17) = 9.423, P = 0.0007; rearing, F(3,17) = 0.8719, P = 0.4749; digging, F(3,17) = 0.2490, P = 0.8609. Female (i–k). WT versus Oxtr-KO (−/−) versus Oxtr-KO (+/−) versus Oxtr-KO (+/−) + LpEV. One-way ANOVA, Tukey’s post-hoc test. Grooming, F(3,12) = 4.502, P = 0.0245; rearing, F(3,12) = 3.064, P = 0.0692; digging, F(3,12) = 0.4702, P = 0.7086. l–n, RT-PCR analysis of Oxt, Avp and Nr4a3 expression in the dStr, NAc and dHP of Oxtr+/− mice after LpEV treatment. n = 4 animals per group; 4 repeats per group. WT versus Oxtr-KO (+/−) versus Oxtr-KO (+/−) + LpEV. One-way ANOVA, Tukey’s post-hoc test. NAc (l): Oxt, F(2,9) = 26.15, P = 0.0002; Avp, F(2,9) = 13.51, P = 0.0019; Nr4a3, F(2,9) = 2.348, P = 0.1511. dStr (m): Oxt, F(2,9) = 16.57, P = 0.0010; Avp, F(2,8) = 2.927, P = 0.1112; Nr4a3, F(2,9) = 3.725, P = 0.0663. dHP (n): Oxt, F(2,9) = 7.423, P = 0.0125; Avp, F(2,9) = 19.50, P = 0.0005; Nr4a3, F(2,9) = 4.206, P = 0.0513. o–q, RT-PCR analysis of Oxt, Oxtr, Avp and Nr4a3 expression in the dStr, NAc and dHP of Drd2-KO mice after LpEV treatment. n = 4 animals per group; ~4–6 repeats per group. WT versus Oxtr-KO (+/−) versus Oxtr-KO (+/−) + LpEV. One-way ANOVA, Tukey’s post-hoc test. NAc (o): Oxt, F(2,10) = 13.19, P = 0.0016; Oxtr, F(2,11) = 13.69, P = 0.0010; Avp, F(2,9) = 33.49, P < 0.0001; Nr4a3, F(2,9) = 1.993, P = 0.1921. dStr (p): Oxt, F(2,13) = 20.61, P < 0.0001; Oxtr, F(2,11) = 3.297, P = 0.0755; Avp, F(2,15) = 63.59, P < 0.0001; Nr4a3, F(2,15) = 9.502, P = 0.0022. dHP (q): Oxt, F(2,15) = 2.908, P = 0.0856; Oxtr, F(2,12) = 6.850, P = 0.0104; Avp, F(2,15) = 8.754, P = 0.0030; Nr4a3, F(2,15) = 5.093, P = 0.0205.

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