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. 2008 Mar;22(3):546–558. doi: 10.1210/me.2007-0264

Fig. 1.

Fig. 1.

BDNF-Enhanced Outgrowth of Neurites Was Suppressed in the Presence of DEX

A, Cultured hippocampal neurons at DIV4 were immunostained with anti-MAP2 (green, neuronal dendritic marker) and anti-GAD (red, GABAergic neuronal marker) antibodies. a, MAP2 image; b, MAP2 and GAD merged image. Approximately 10% of MAP2-positive cells were GAD-positive GABAergic neurons in our cultures. Bar, 20 μm. B, Experimental schedule of BDNF and DEX application for immunostaining. DEX was applied at DIV1, and then BDNF was also applied at DIV2 in the presence of DEX. Forty-eight hours after BDNF addition, the cultured cells were fixed for immunostaining. C, Influence of DEX treatment on neurite outgrowth of GABAergic neurons. a, d, g, and j, The neurites were revealed by immunostaining with anti-MAP2 antibody; b, e, h, and k, GABAergic neurons were determined by staining with anti-GAD antibody; c, f, i, and l, merged images; d–f, BDNF has a slight tendency to increase the number of neurites compared with control (a–c) (none), although significance was not observed; g–i, DEX pretreatment before BDNF application; j–l, sole DEX treatment. Bar, 20 μm. m, The number of neurites per cell was counted. Data represent mean ± sd (none, n = 60; BDNF, n = 58; BDNF plus DEX, n = 73; DEX, n = 48). The n indicates the number of cells. D, Effect of DEX on neurite outgrowth of glutamatergic neurons. MAP2-positive (a, d, g, and j) and glutamate-positive (b, e, h, and k) glutamatergic neurons are shown. c, f, i, and l, Merged images; d–f, BDNF increased the number of neurites compared with control (a–c); g–i, DEX significantly suppressed the BDNF effect; j–l, sole DEX had no influence compared with control. Bar, 20 μm. m, Quantification indicates that the number of glutamatergic neurites was increased by BDNF, and the increase was suppressed by DEX. Data represent mean ± sd (none, n = 200; BDNF, n = 200; BDNF plus DEX, n = 200; DEX, n = 200). The n indicates the number of cells. Reproducibility was confirmed by three independent cultures. Statistical significance was determined by ANOVA. ***, P < 0.001; **, P < 0.01.