XB-IMG-127276
Xenbase Image ID: 127276
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Figure 2. Transgenic expression of Venus-GABAARα1.(A) Strategy for Otx1 BAC modification with VGABAARα1. The red line shows the Southern blot probe used in (B). Scale: 2 kb. (B) Correct incorporation of Venus-GABRA1 cDNA into the Otx1 BAC is shown by southern blotting. The modified BAC (middle lane) contains an additional EcoR1 site. The right lane shows correct incorporation of the modified BAC into the mouse genome. The transgenic mouse genome contains a wild-type copy of the Otx1 regulatory region as well as the modified Otx1-Venus-GABRA1 BAC. (C) Cortical protein extract from wild type and Otx1-VGABAARα1 mice immunoblotted with anti-GABAARα1 antibody. Only the transgenic mouse expresses the fusion version of the GABAARα1 subunit (top band). (D) VGABAARα1 expression in cortical layers 5 and 6 pyramidal neurons of Otx1-VGABAARα1 mice is shown by GFP immunoreactivity. The fusion protein is localized to pyramidal cell soma in layers 5/6 and processes in layers 2/3. Scale: 500 µm. (E) Immunofluorescence shows the colocalization of VGABAARα1 (green) and NeuN (red), a neuronal marker, in layers 5 and 6 pyramidal neurons of Otx1- VGABAARα1 transgenic mice (left). VGABAARα1 is mainly localized to the perikarya of the cell soma as well as dendrites. A control Otx1 BAC transgenic mouse expresses soluble eGFP (right), which fills the cell soma. Scale: 100 µm. V: Venus. GAR: GABAA receptor. Image published in: Heller EA et al. (2012) Heller et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |