PMC:1255741 / 65816-68769 JSONTXT

{"project":"2_test","denotations":[{"id":"16216087-11967539-84723805","span":{"begin":144,"end":146},"obj":"11967539"},{"id":"16216087-9916792-84723806","span":{"begin":232,"end":234},"obj":"9916792"},{"id":"16216087-12195431-84723807","span":{"begin":1227,"end":1229},"obj":"12195431"},{"id":"T3366","span":{"begin":144,"end":146},"obj":"11967539"},{"id":"T37591","span":{"begin":232,"end":234},"obj":"9916792"},{"id":"T45082","span":{"begin":1227,"end":1229},"obj":"12195431"}],"text":"Supporting Information\nFigure S1 Forebrain-Specific Gene Manipulation\n(A) Schematic diagrams depicting Cre under the αCaMKII promoter control [59] and Cre-dependent expression of β-galactosidase (LacZ) by the R26R indicator mouse [85].\n(B) Coronal forebrain sections (left) and sagittal brainstem/cerebellum sections of mice at P42 positive for TgCre4 and R26R Cre-indicator. Cre expression visualized by enzymatic β-galactosidase activity (blue, X-gal, counterstain by eosin) of R26R and by anti-Cre immunostainings (brown, DAB [diaminobenzadine]) was restricted to forebrain regions. Scale bars: 1 mm. A, amygdala; Ce, Cerebellum; Cx, cortex; H, hippocampus; Me, medulla oblongata.\n(C) Olfactory bulb sections of the same mouse to visualize Cre expression by Cre immunoreactivity (left, DAB) and by enzymatic β-galactosidase activity (right, X-gal, counterstain by eosin) in granule cells (GC) and mitral cells (MC) as indicated by arrows. Scale bars: 400 μm (upper panel), 50 μm (lower panel).\n(3.4 MB TIF).\nClick here for additional data file.\nFigure S2 Performance of GluR-BΔFB (n = 10) and Littermate Control (n = 10) Mice in a Spatial Reference Task on the Elevated Y-Maze\nDetails of the methodology are described in [90]. * indicates p \u003c 0.05.\n(126 KB TIF).\nClick here for additional data file.\nFigure S3 Memory Deficit Is Not Due to Increased Extinction\n(A) Memory performance as a function of time for the experiment described in Figure 2E (nine GluR-BΔFB and nine GluR-B2lox control animals). Only four unrewarded “memory +” trials are binned for each data point. Memory of GluR-BΔFB animals was significantly reduced (F(1,33) = 17; p \u003c 0.001). Whereas a weak overall time effect could be observed (F(2,66) = 3.6; p = 0.03), there was no genotype-time interaction effect (F(2,66) = 0.67; p = 0.5), indicating that there is no differential effect of putative extinction on memory performance. * indicates p-values for a Mann-Whitney U test (* \u003c 0.05, ** \u003c 0.01).\n(B) After the memory experiments from Figure 6A, the last set of animals (eight GluR-BRescue, four GluR-BΔFB, and three controls) was further trained on AA versus EB for 900 trials one week after the memory experiment. Subsequently, training continued on AA/EB mixtures for 1,200 trials. Finally, the animals were retrained on AA and EB for 100 trials. Relearning performance during the last retraining task is highly correlated to the original memory performance (R2 = 0.46, p = 0.006, n = 15).\n(264 KB TIF).\nClick here for additional data file.\nFigure S4 Absence of Cre Expression in the Main Olfactory Epithelium of TgCre4 Mice\n(A) Overview of the main olfactory epithelium. (B) Higher magnification of (A) showing all cells with propidium iodide (red) and absence of Cre-positive cells (green, anti-Cre). (C) Positive control showing granule cells of the dentate gyrus stained for nuclear Cre (green) from the same mouse.\n(3.7 MB TIF).\nClick here for additional data file."}