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. 2021 May 31;12:3321. doi: 10.1038/s41467-021-23843-0

Fig. 5. P2X2 receptors in the prefrontal cortex mediate the beneficial effects of ATP on ASD-like behaviors.

Fig. 5

a A schematic of the AAV vector carrying P2X2R-shRNA (top). The viral expression of P2X2R-shRNA (bottom right) and the injection site in the mPFC (bottom left, red box, scale bar: 100 µm). b Western blot showing P2X2R knockdown in the mPFC with AAV-P2X2R-shRNA (two-tailed unpaired t-test, t6 = 12.71, P = 0.15 × 10−4, n = 4). c, d i.p. injection of ATP (125 mg/kg) rescued social interaction in IP3R2 cKO mice, and this effect was blocked by P2X2R knockdown (c F7,45 = 6.973, P = 0.7 × 10−5; Con:Control-shRNAvehicle vs. Con:P2X2R-shRNAvehicle, P = 0.0142; Con:Control-shRNAvehicle vs. cKO:Control-shRNAvehicle, P = 0.0066; Con:Control-shRNAvehicle vs. cKO:P2X2R-shRNAvehicle, P = 0.0002; Con:P2X2R-shRNAvehicle vs. Con:P2X2R-shRNAATP, P = 0.8171; cKO:Control-shRNAvehicle vs. cKO:Control-shRNAATP, P = 0.0037; cKO:P2X2R-shRNAvehicle vs. cKO:P2X2R-shRNAATP, P = 0.8931; d F7,45 = 0.8109, P = 0.5830; n = 6,7,7,7,6,7,7,7). e sIPSC recordings with and without ATPγS (25 µM) treatment in mPFC layer 5 pyramidal neurons from control and IP3R2 cKO mice injected with control-shRNA or P2X2R-shRNA. f, g Bar graphs showing sIPSC frequency (f Con:Control-shRNAACSF vs. Con:P2X2R-shRNAACSF, two-tailed unpaired t-test, t14 = 2.381, P = 0.0320, n = 10/6; Con:Control-shRNAACSF vs. cKO:control-shRNAACSF, two-tailed unpaired t-test, t18 = 3.539, P = 0.0023, n = 10; Con:Control-shRNAACSF vs. cKO:P2X2R-shRNAACSF, two-tailed unpaired t-test, t17 = 3.273, P = 0.0045, n = 9/10; Con:P2X2R-shRNAACSF vs. Con:P2X2R-shRNAATPγs, two-tailed paired t-test, t5 = 2.001, P = 0.1018, n = 6; cKO:Control-shRNAACSF vs. cKO:Control-shRNAATPγs, two-tailed paired t-test, t9 = 2.777, P = 0.0215, n = 10; cKO:P2X2R-shRNAACSF vs. cKO:P2X2R-shRNAATPγs, two-tailed paired t-test, t8 = 0.1624, P = 0.8750, n = 9) and amplitude (g) with or without ATPγS (25 µM) treatment in mPFC layer 5 pyramidal neurons from control and IP3R2 cKO mice injected with control-shRNA or P2X2R-shRNA. P2X2R knockdown prevented the enhancing effect of ATPγS (25 µM) on sIPSC frequency in IP3R2 cKO slices. Scale bars: 20 pA, 2 s. Each data point represents an individual mouse (c, d). Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. ns, no significant difference. One-way ANOVA with Fisher’s LSD multiple comparison post hoc test (c, d). Source data are provided as a Source data file.