FIGURE 2.

Generation of tamoxifen-inducible Thy1-Cre mice with GCS deletion in subsets of adult forebrain neurons. (A) The targeting construct in Thy1-CreERT2/EYFP mice comprises one Thy1 copy that drives tamoxifen-inducible Cre recombinase and the second Thy1 copy that drives tamoxifen-independent EYFP in targeted neurons. (B) EYFP fluorescence shows that neurons in the hippocampal regions CA1, CA2, and dentate gyrus are targeted by the construct (bregma –3.08 mm (coronal), scale bars = 200 μm). (C) An in situ hybridization (ISH) of tamoxifen-induced Ugcgf/f//Thy1-CreERT2/EYFP (Thy1-Cre) mice confirms that Cre-targeted neurons are devoid of GCS expression. The original brown ISH dots have been converted to red fluorescence, as described in Materials and Methods section (scale bar = 10 μm). For comparison, the original images are depicted in Supplementary Figure 2C. (D) Immunofluorescence shows that ganglioside GD1a is absent in Cre-targeted and fluorescent neurons of tamoxifen-induced Thy1-Cre mice. On the contrary, non-induced Thy1-Cre mice (n.i. control) mice only receiving solvent injections without tamoxifen display GD1a expression in EYFP-fluorescent neurons, because Cre activity is not induced in this case (scale bar = 5 μm).