aav expressing egfp cre recombinase  (Addgene inc)

Journal: Cells

Article Title: Perturbation of Cortical Excitability in a Conditional Model of PCDH19 Disorder

doi: 10.3390/cells11121939

Figure Lengend Snippet: ( A ) A focal mosaic of PCDH19 expression was created by injecting an AAV expressing EGFP-Cre recombinase into the right visual cortex (V1) of the PCDH19flox/flox mouse. The opposite, non-injected hemisphere was used as the internal control. ( B ) PCDH19 mosaic mice experienced a transient period of unexpected mortality. Data collected from 70 mice injected at P1 (blue) differs significantly from that of control littermates (N = 48 mice, black; Mantel-Cox test p < 0.01). Yellow area indicates the period of adolescence in mice. ( C ) PCDH19 mice have a normal diurnal alternation of resting phases during the day (light on) and of high locomotor activity during the night (light off, shaded area; darkness from 7 PM till 7 AM). Bars represent the speed averaged in 5 min bins measured in three control and three PCDH19 mice in a 48-h period. Recordings start at 12 AM. ( D ) Density maps showing the arena occupation during a 2-h period of the active phase. ( E ) PCDH19 mice are hyperactive during the night phase. ( F ) PCDH19 mice spend less time than controls in resting states that can be ascribed to sleep (N = 8 for both control and PCDH19 mice; data pooled from 36 and 38 days of video recording). Significant differences are indicated with asterisks (** p < 0.02, Mann–Whitney test).

Article Snippet: Mosaic PCDH19 expression was induced in the occipital cortex of PCDH19 conditional knockout mice by focal injection of an AAV expressing EGFP-Cre recombinase (Addgene #105545-AAV1; pAAV.CMV.HI.eGFP-Cre.WPRE.SV40).

Techniques: Expressing, Injection, Control, Activity Assay, MANN-WHITNEY

Journal: Cells

Article Title: Perturbation of Cortical Excitability in a Conditional Model of PCDH19 Disorder

doi: 10.3390/cells11121939

Figure Lengend Snippet: In vivo 2-photon calcium imaging in PCDH19 mosaic brain. ( A ) Example field of view with Cre-negative red neurons (top grey square; expressing jRGECO1a only) and Cre positive (PCDH19 knockout) green neurons (bottom square) that express jRGECO1a in the cytoplasm and EGFP-Cre recombinase in the nucleus. ( B ) Calcium transients (green trace) measured over the entire field of view. The vertical axis displays the number of pixels affected by calcium activity over time (see Supplemental Information). The black trace shows the LFP recorded from the same hemisphere with an electrode positioned just outside of the imaged field at a depth of 250 µm. The inset shows magnified traces in correspondence of the black bar. Notice how Ca 2+ transients are phase locked to the USs. ( C ) Frequency of calcium transients in red and green cells of mosaic mice and controls. Each dot represents the average value for each active neuron. Significant differences are indicated with asterisks (*** p < 0.0005, ** p < 0.005; Mann–Whitney test). Transient frequency is significantly increased in mosaics compared to control mice both for Cre+ (PCDH19 KO) neurons and for Cre- neurons. ( D ) Box plots of the percentage of Ca 2+ transients occurring within USs, demonstrating a significant reduction in synchronization with the SWA oscillation in mosaic mice compared to control mice (* p < 0.02; Mann–Whitney test, whiskers indicate data range). Each dot represents the value obtained by all neurons in the imaging field of PCDH19 mosaic mice. The filled magenta dots represent the values obtained from mosaic animals with a hyperexcitable (>5 β oscillations per hour) phenotype, the blue dots from all other mosaics.

Article Snippet: Mosaic PCDH19 expression was induced in the occipital cortex of PCDH19 conditional knockout mice by focal injection of an AAV expressing EGFP-Cre recombinase (Addgene #105545-AAV1; pAAV.CMV.HI.eGFP-Cre.WPRE.SV40).

Techniques: In Vivo, Imaging, Expressing, Knock-Out, Activity Assay, MANN-WHITNEY, Control