Figure 3.
Increased PI(3)P synthesis upon cLTP regulates the formation of SNX17- and SNX27-positive puncta. (A) Representative confocal images of DIV17 hippocampal neurons that were transfected at DIV16 with GFP-SNX17 and dsRed-EEA1-FYVE. 24 h later, neurons were either fixed in the absence of cLTP (baseline) or treated with cLTP for 5 min and fixed 10 or 30 min after the cLTP stimulus. Scale bar, 5 µm. (B) The number of dsRed-EEA1-FYVE–positive puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.241 ± 0.010, N = 30 neurons; cLTP 10 min: 0.327 ± 0.013, N = 30 neurons; cLTP 30 min: 0.361 ± 0.010, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (C) The number of GFP-SNX17 puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.227 ± 0.009, N = 30 neurons; cLTP 10 min: 0.317 ± 0.013, N = 30 neurons; cLTP 30 min: 0.349 ± 0.010, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (D) The colocalization between dsRed-EEA1-FYVE– and GFP-SNX17–positive puncta was analyzed using Mander’s colocalization coefficient (×100). Baseline: 84.25 ± 2.019%, N = 30 neurons; cLTP 10 min: 84.94 ± 2.041%, N = 30 neurons; cLTP 30 min: 82.87 ± 1.751%, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Error bars are SEM. (E) Representative confocal images of DIV17 hippocampal neurons that were transfected at DIV16 with GFP-SNX27 and dsRed-EEA1-FYVE. 24 h later, neurons were either fixed in the absence of cLTP (baseline) or treated with cLTP for 5 min and fixed 10 or 30 min after the cLTP stimulus. Scale bar, 5 µm. (F) The number of dsRed-EEA1-FYVE–positive puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.268 ± 0.010, N = 30 neurons; cLTP 10 min: 0.353 ± 0.013, N = 30 neurons; cLTP 30 min: 0.389 ± 0.015, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (G) The number of GFP-SNX27 puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.258 ± 0.010, N = 30 neurons; cLTP 10 min: 0.340 ± 0.013, N = 30 neurons; cLTP 30 min: 0.378 ± 0.014, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (H) The colocalization between dsRed-EEA1-FYVE– and GFP-SNX27–positive puncta was analyzed using Mander’s colocalization coefficient (×100). Baseline: 84.60 ± 1.668%, N = 30 neurons; cLTP 10 min: 86.17 ± 1.483%, N = 30 neurons; cLTP 30 min: 85.68 ± 2.027%, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Error bars are SEM. DIV, days in vitro.

Increased PI(3)P synthesis upon cLTP regulates the formation of SNX17- and SNX27-positive puncta. (A) Representative confocal images of DIV17 hippocampal neurons that were transfected at DIV16 with GFP-SNX17 and dsRed-EEA1-FYVE. 24 h later, neurons were either fixed in the absence of cLTP (baseline) or treated with cLTP for 5 min and fixed 10 or 30 min after the cLTP stimulus. Scale bar, 5 µm. (B) The number of dsRed-EEA1-FYVE–positive puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.241 ± 0.010, N = 30 neurons; cLTP 10 min: 0.327 ± 0.013, N = 30 neurons; cLTP 30 min: 0.361 ± 0.010, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (C) The number of GFP-SNX17 puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.227 ± 0.009, N = 30 neurons; cLTP 10 min: 0.317 ± 0.013, N = 30 neurons; cLTP 30 min: 0.349 ± 0.010, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (D) The colocalization between dsRed-EEA1-FYVE– and GFP-SNX17–positive puncta was analyzed using Mander’s colocalization coefficient (×100). Baseline: 84.25 ± 2.019%, N = 30 neurons; cLTP 10 min: 84.94 ± 2.041%, N = 30 neurons; cLTP 30 min: 82.87 ± 1.751%, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Error bars are SEM. (E) Representative confocal images of DIV17 hippocampal neurons that were transfected at DIV16 with GFP-SNX27 and dsRed-EEA1-FYVE. 24 h later, neurons were either fixed in the absence of cLTP (baseline) or treated with cLTP for 5 min and fixed 10 or 30 min after the cLTP stimulus. Scale bar, 5 µm. (F) The number of dsRed-EEA1-FYVE–positive puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.268 ± 0.010, N = 30 neurons; cLTP 10 min: 0.353 ± 0.013, N = 30 neurons; cLTP 30 min: 0.389 ± 0.015, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (G) The number of GFP-SNX27 puncta in the first 30 μm of secondary dendrites was quantified at the indicated time points. Baseline: 0.258 ± 0.010, N = 30 neurons; cLTP 10 min: 0.340 ± 0.013, N = 30 neurons; cLTP 30 min: 0.378 ± 0.014, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test, ****P < 0.001. Error bars are SEM. (H) The colocalization between dsRed-EEA1-FYVE– and GFP-SNX27–positive puncta was analyzed using Mander’s colocalization coefficient (×100). Baseline: 84.60 ± 1.668%, N = 30 neurons; cLTP 10 min: 86.17 ± 1.483%, N = 30 neurons; cLTP 30 min: 85.68 ± 2.027%, N = 30 neurons. Three independent experiments. Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. Error bars are SEM. DIV, days in vitro.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal