Neurturin Evokes MAPK-Dependent Upregulation of Egr4 and KCC2 in Developing Neurons
Regulation of KCC2 expression by Neurturin in vivo. (a) RET and GFRα2 mRNA expression was detected by in situ hybridization. Note the accumulation of RET and GFRα2 at P3 and P5 in the CA3 region of the hippocampus (marked with arrowheads). Scale bar = 500 μm. (b) Representative immunofluorescent stainings of KCC2 expression in Neurturin-treated and control hippocampi 3 days after injection. Neurturin (1 μg) was used for injection in right-side hippocampus of P5 rat while contralateral hippocampus was injected with saline solution. (c) Summary results of KCC2 immunostaining intensity for different layers of CA1 and CA3 in Neurturin-injected hemisphere (n = 16–21). KCC2 intensity was measured in 6 sections at various distances from injection site. The effect of Neurturin injection was uneven along the dorsal-ventral axis. Quantification was made for the following layers: CA3 is whole area of CA3; pCA3 and pCA1 are pyramidal layers of CA3 and CA1, respectively; SO is stratum oriens; SR is stratum radiatum and LM is stratum lacunosum-moleculare. In each section and each layer KCC2 intensity in Neurturin-injected hemisphere was normalized to the corresponding value in control hemisphere. Then for each layer the maximum, the minimum, and the average values of KCC2 intensity along the dorsal-ventral axis were calculated (**, ***). Error bars represent SEM. Scale bar = 300 μm.
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