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. 2018 Jan 3;38(1):183–199. doi: 10.1523/JNEUROSCI.1640-17.2017

Figure 2.

Figure 2.

PKD1 is essential for synapse formation and function. A, Representative images and quantification of spine density and area in DIV 15 hippocampal neurons cotransfected GFP with his-myc-tagged Vector, hPKD1, or DN-hPKD1 at DIV 8 (n = 20, 16, 16 cells, respectively, in columns shown in the graphs). Scale bars: Top, 50 μm; Bottom, 5 μm. One-way ANOVA with Bonferroni's post hoc test: ***p < 0.001, compared with Vector. ###p < 0.001, compared with hPKD1. B, Representative traces (B1) and plots (B2) of mEPSC frequencies and amplitudes in hippocampal neurons cotransfected GFP with Vector, hPKD1, or DN-hPKD1 (n = 10, 10, 11 cells, respectively). Calibration: 10 pA, 500 ms. One-way ANOVA with Bonferroni's post hoc test: *p < 0.05, compared with Vector. **p < 0.01, compared with Vector. ###p < 0.001, compared with hPKD1. No significant differences in mEPSC amplitude were detected. C, Knockdown effect of myc-tagged rat PKD1 (rat-hPKD1) by shPKD1 in N2a cells. Rat-hPKD1 is an analog of hPKD1 with the rat target sequence of shPKD1 replacing the homologous domain in hPKD1 by site-directed mutagenesis. D, Representative images and quantification of spine density and area of DIV 15 hippocampal neurons transfected with shGFP+Vector, shPKD1+Vector, shPKD1+hPKD1, or shPKD1+DN-hPKD1 at DIV 8 (n = 18, 17, 15, and 16 cells, respectively). Scale bar, 2.5 μm. One-way ANOVA with Bonferroni's post hoc test: *p < 0.05, compared with shGFP+Vector. **p < 0.01, compared with shGFP+Vector. ***p < 0.001 compared with shGFP+Vector. ###p < 0.001, compared with shPKD1+Vector. E, Representative traces (E1) and plots (E2) of the mEPSCs in hippocampal neurons transfected with shGFP+Vector, shPKD1+Vector, shPKD1+hPKD1, or shPKD1+DN-hPKD1 (n = 10, 11, 7, and 7 cells, respectively). Calibration: 10 pA, 500 ms. One-way ANOVA with Bonferroni's post hoc test: **p < 0.01, compared with shGFP+Vector. ***p < 0.001, compared with shGFP+Vector. #p < 0.05 compared with shPKD1+Vector. No significant differences in mEPSC amplitude were detected among the groups. Data are mean ± SEM.