Figure 3.
KIF13A domain analysis for LTP. (A) Time course (mean of the normalized excitatory postsynaptic currents [EPSCs] ± SEM per condition) of the LTP induction in organotypic hippocampal slices (7‒10 DIV) expressing GFP-KIF13A-ST (green) or mRFP-KIF13A-tail (orange) at 24 h after infection. Top left: Inset shows schematic representation of the GFP-KIF13A-ST and mRFP-KIF13A-tail recombinant proteins with known domains (coiled-coil [C-C] and FHA). Top right: Insets show representative traces for uninfected (black lines), GFP-KIF13A-ST (green lines), and mRFP-KIF13A-tail (orange lines) neurons, averaged from baseline (dark lines) or from the last 5 min of the recording (light lines). Scale bars: vertical, 20 pA; horizontal, 10 ms. Bars show mean ± SEM, together with individual values for each experiment, from the last 5 min of the recording and normalized to the baseline. n = 19, 11, or 9 cells for control, GFP-KIF13A-ST– or mRFP-KIF13A-tail–expressing neurons, respectively, for the LTP (paired) pathway. n = 12, 5, or 5 cells for control, GFP-KIF13A-ST–,or mRFP-KIF13A-tail–expressing neurons, respectively, for the control (unpaired) pathway. Uninfected and KIF13A-tail–expressing neurons are significantly potentiated (P = 0.002 and P = 0.01, respectively) with respect to baseline (Wilcoxon test). P values displayed in the panel represent KIF13A-ST significant difference from uninfected (P = 0.001) and KIF13A-tail (P = 0.002), respectively (Mann–Whitney test). (B and C) Basal synaptic responses mediated by AMPARs (B) and NMDARs (C) from organotypic hippocampal slices (7‒10 DIV) infected with a Sindbis virus expressing GFP-KIF13A-ST. Synaptic responses were recorded from pairs of uninfected and infected neighboring CA1 neurons. Gray circles represent values from each individual pair, while green symbols show the mean ± SEM of the measured currents. n = 15 cell pairs for AMPAR currents and n = 12 cell pairs for NMDAR currents; n.s., not significantly different (Wilcoxon test). Insets: Representative traces for uninfected (black lines) and GFP-KIF13A-ST (green lines) cells; scale bars: vertical, 10 pA; horizontal, 10 ms. (D) Time course (mean of the normalized EPSCs ± SEM per condition) of the LTP induction in organotypic hippocampal slices (7‒9 DIV) transfected with a biolistic approach to allow simultaneous expression of shKIF13A and KIF13A-MS-YFP (pink; KIF13A-MS-rescue) for 7–9 d. Top left: Inset shows schematic representation of the KIF13A-MS-YFP recombinant protein. Top right: Insets show representative traces for uninfected (black lines) and KIF13A-MS-rescue (pink lines) neurons, averaged from baseline (dark lines) or from the last 5 min of the recording (light lines). Scale bars: vertical, 10 pA; horizontal, 10 ms. Bars show mean ± SEM, together with individual values for each experiment, from the last 10 min of the recording and normalized to the baseline. n = 9 uninfected cells and n = 10 KIF13A-MS rescue neurons for the LTP (paired) pathway and n = 6 uninfected cells and n = 5 KIF13A-MS rescue neurons for the control (unpaired) pathway. Uninfected and KIF13A-MS-rescue neurons are significantly potentiated (P = 0.008 and P = 0.02, respectively) with respect to baseline (Wilcoxon test).

KIF13A domain analysis for LTP. (A) Time course (mean of the normalized excitatory postsynaptic currents [EPSCs] ± SEM per condition) of the LTP induction in organotypic hippocampal slices (7‒10 DIV) expressing GFP-KIF13A-ST (green) or mRFP-KIF13A-tail (orange) at 24 h after infection. Top left: Inset shows schematic representation of the GFP-KIF13A-ST and mRFP-KIF13A-tail recombinant proteins with known domains (coiled-coil [C-C] and FHA). Top right: Insets show representative traces for uninfected (black lines), GFP-KIF13A-ST (green lines), and mRFP-KIF13A-tail (orange lines) neurons, averaged from baseline (dark lines) or from the last 5 min of the recording (light lines). Scale bars: vertical, 20 pA; horizontal, 10 ms. Bars show mean ± SEM, together with individual values for each experiment, from the last 5 min of the recording and normalized to the baseline. n = 19, 11, or 9 cells for control, GFP-KIF13A-ST– or mRFP-KIF13A-tail–expressing neurons, respectively, for the LTP (paired) pathway. n = 12, 5, or 5 cells for control, GFP-KIF13A-ST–,or mRFP-KIF13A-tail–expressing neurons, respectively, for the control (unpaired) pathway. Uninfected and KIF13A-tail–expressing neurons are significantly potentiated (P = 0.002 and P = 0.01, respectively) with respect to baseline (Wilcoxon test). P values displayed in the panel represent KIF13A-ST significant difference from uninfected (P = 0.001) and KIF13A-tail (P = 0.002), respectively (Mann–Whitney test). (B and C) Basal synaptic responses mediated by AMPARs (B) and NMDARs (C) from organotypic hippocampal slices (7‒10 DIV) infected with a Sindbis virus expressing GFP-KIF13A-ST. Synaptic responses were recorded from pairs of uninfected and infected neighboring CA1 neurons. Gray circles represent values from each individual pair, while green symbols show the mean ± SEM of the measured currents. n = 15 cell pairs for AMPAR currents and n = 12 cell pairs for NMDAR currents; n.s., not significantly different (Wilcoxon test). Insets: Representative traces for uninfected (black lines) and GFP-KIF13A-ST (green lines) cells; scale bars: vertical, 10 pA; horizontal, 10 ms. (D) Time course (mean of the normalized EPSCs ± SEM per condition) of the LTP induction in organotypic hippocampal slices (7‒9 DIV) transfected with a biolistic approach to allow simultaneous expression of shKIF13A and KIF13A-MS-YFP (pink; KIF13A-MS-rescue) for 7–9 d. Top left: Inset shows schematic representation of the KIF13A-MS-YFP recombinant protein. Top right: Insets show representative traces for uninfected (black lines) and KIF13A-MS-rescue (pink lines) neurons, averaged from baseline (dark lines) or from the last 5 min of the recording (light lines). Scale bars: vertical, 10 pA; horizontal, 10 ms. Bars show mean ± SEM, together with individual values for each experiment, from the last 10 min of the recording and normalized to the baseline. n = 9 uninfected cells and n = 10 KIF13A-MS rescue neurons for the LTP (paired) pathway and n = 6 uninfected cells and n = 5 KIF13A-MS rescue neurons for the control (unpaired) pathway. Uninfected and KIF13A-MS-rescue neurons are significantly potentiated (P = 0.008 and P = 0.02, respectively) with respect to baseline (Wilcoxon test).

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