Neural Plasticity / 2011 / Article / Fig 1

Research Article

Neurturin Evokes MAPK-Dependent Upregulation of Egr4 and KCC2 in Developing Neurons

Figure 1

Regulation of KCC2 expression by Neurturin. (a) Representative Western blot analysis of Neurturin-induced ERK1/2 phosphorylation. Lysates of div5-dissociated cultures were collected 5–20 min after Neurturin (50 ng/mL) application. In some cases cultures were pretreated with MEK inhibitor U0126 (20 μM) 30 min before Neurturin application. (b) Egr4 mRNA level in div5–div10 dissociated cultures 1-2 hours after Neurturin (50 ng/mL) application as detected by real time PCR ( 𝑛 = 5 ). Nontreated control value was set to 1. (* 𝑃 < 0 . 0 5 , ** 𝑃 < 0 . 0 1 , *** 𝑃 < 0 . 0 0 1 ). Error bars represent SEM. (c) Representative Western blot analysis and quantification of KCC2 expression in organotypic hippocampal slice cultures treated with NTRN (n = 4–8). Organotypic cultures were treated with 10 ng/mL and 50 ng/mL Neurturin. Data are normalized to the value in nontreated controls (* 𝑃 < 0 . 0 5 , ** 𝑃 < 0 . 0 1 , *** 𝑃 < 0 . 0 0 1 ). Error bars represent SEM. (d) Representative Western blot analysis and quantification of KCC2 expression in dissociated hippocampal cultures treated with Neurturin (50 ng/mL) (n = 3–7). Dissociated cultures were treated with Neurturin at div1, div8 and div15 and analyzed 3 days after the treatment. Data are normalized to the value of non-treated controls of the corresponding age (* 𝑃 < 0 . 0 5 , ** 𝑃 < 0 . 0 1 , *** 𝑃 < 0 . 0 0 1 ). Error bars represent SEM. (e) Representative semiquantitative RT PCR from cDNA of different age cultures for RET, GFRα2, and GAPDH (used as internal standard) and summarized results of 3 similar PCRs. The data show that dissociated hippocampal neurons express detectable levels of growth factors receptors at all ages tested. Data are normalized to div4 value. Error bars represent SEM.
641248.fig.001a
(a)
641248.fig.001b
(b)
641248.fig.001c
(c)
641248.fig.001d
(d)
641248.fig.001e
(e)

Article of the Year Award: Outstanding research contributions of 2020, as selected by our Chief Editors. Read the winning articles.