Research Article

The Activator Mastoparan-7 Promotes Dendritic Spine Formation in Hippocampal Neurons

Figure 1

Mas-7 activates the subunit and increases the intracellular Ca2+ concentration in hippocampal neurons. (a) Left panel, 14 DIV hippocampal neurons were stimulated with 1 μM Mas-7 for 0, 5, or 30 min. The neurons were lysed and incubated with anti--GTP for 2 h and then analyzed by immunoblotting using an anti- antibody (). The input lane corresponds to a lysate sample before the immunoprecipitation. . Right panel, lysates from untreated (control) hippocampal neurons were incubated with GTPγS as a positive control or with GDP as a negative control for 90 min at RT. Then, the -GTP was immunoprecipitated and analyzed by western blotting to determine the total level of . The IgG band shows that an equal amount of antibody was used for the immunoprecipitation. (b) Representative western blot and quantification of the total level of in 14 DIV neurons incubated for different periods of time with 1 μM Mas-7. GAPDH was used as a loading control (). (c) Ratio images (340/380) of the Fura-2AM probe from hippocampal neurons under basal conditions () or after 3 min of 1 μM Mas-7 treatment. (d) Quantification of measurements of the intracellular Ca2+ increase in hippocampal neurons bathed in a Ca2+-free solution with different concentrations of Mas-7 (, 70–79 neurons, each condition). (e) Area under the curve of the Ca2+ increase after Mas-7 treatment. .