Generation and confirmation of transgenic mice overexpressing the KCNQ2 dominant-negative pore mutant in forebrain. A. Schematic organization of the transgene for forebrain-specific expression of the dominant-negative mutant of rat KCNQ2-G279S (rQ2-G279S) driven by the αCaMKII promoter. B. Identification of rQ2-G279S mRNA in the brain isolated from transgenic (Tg) and wild type (WT) mice by real time RT-PCR (mean ± s.e.m., n = 4, *p < 0.05). The template of rQ2-G279S specific primers for detection of KCNQ2 mutant gene expression is located in the area of SV40 poly A of the 265-plasmid (transgenic construct). The template of genomic DNA primers for detection of genomic DNA contamination is located after SV40 poly A in the 265-plasmid (transgenic construct), which does not undergo transcription. β-actin is used as an internal reference. C. Expression of rQ2-G279S mRNA isolated from 5 different tissues of WT and Tg mice by real time RT-PCR. The transgene rQ2-G279S was adequately expressed in the cortex, hippocampus and thalamus, with little expression in the cerebellum, and was not expressed in the DRG of Tg mice (mean ± s.e.m., n = 4, *p < 0.05). D. Expression of rQ2-G279S mRNA of WT (top) and Tg mice (down) using in situ hybridyzation. The transgene rQ2-G279S was adequately expressed in the cortex, hippocampus, and thalamus, with little expression in the cerebellum (crbl) and pons, and was not expressed in the medulla of Tg mice. The scale bar is 2mm. E. Nissl-stained coronal sections of hippocampus. No obvious structural change was found in adult transgenic mice (a) or wild type littermates (b). Scale bar, 500 μm. F. Locomotor activity test, no significant difference was detected in wild type (WT) and transgenic mice (Tg) in the total distance traveled during a period of 30 minutes (mean ± s.e.m., n = 7).