Journal: The Journal of Neuroscience

Article Title: Cerebellar Learning Properties Are Modulated by the CRF Receptor

doi: 10.1523/JNEUROSCI.3106-15.2018

Figure Lengend Snippet: CRFR1cKO mice show enhanced learning in the EBC procedure compared with control mice. a, Schematic illustration of EBC using the MDMT. b, Over the course of EBC, a 330 ms CS (green line) was paired with a co-terminating 30 ms US (blue line). c, d, Heatmap of group averages of trial-by-trial eyelid closure over the entire course of conditioning for controls (c) and CRFR1cKO mice (d). e, f, Averaged group eyelid traces per session depicting levels of eyelid closure over the entire course of conditioning for controls (e) and CRFR1cKO mice (f). g, Although both groups performed CRs before US presentation within several training sessions, quantitative assessment clearly reveals that CRFR1cKO CR percentage presented during EBC training is higher than controls (control: green, CRFR1cKO: purple, N = 6, 10, respectively; p = 0.002). CR timing histograms for session 10 are presented in Figure 6-1. Similar learning trends are seen for CRFR1cKO and control mice trained with 250 ms interstimulus interval in Figure 6-2. Data are represented as mean ± SEM. ***p < 0.005.

Article Snippet: Lysates were subjected to gel electrophoresis on 10% SDS-PAGE and separated proteins were transferred onto nitrocellulose membranes and probed with AB5453 Rabbit Anti-GABA A Receptor α6 Antibody (Merck-Millipore; RRID: AB_177478 ) or Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783; Abcam; RRID: AB_303296 ).

Techniques: Control

Journal: The Journal of Neuroscience

Article Title: Cerebellar Learning Properties Are Modulated by the CRF Receptor

doi: 10.1523/JNEUROSCI.3106-15.2018

Figure Lengend Snippet: Establishment of a mouse line depleted of CRFR1 specifically in cerebellar GrCs. The α6 subunit levels in Δα6-Cre line cerebella and the Cre line granular specificity are presented in Figure 2-1. a, Schematic representation of cell-specific deletion of CRFR1 using the Cre-lox system. b, CRFR1 mRNA is reduced by >90% in the cerebella of Cre-expressing mice (CRFR1cKO; N = 8, 6; p = 0.002; data are presented as fold change). c, No significant reduction in CRFR1 mRNA levels was detected in extracerebellar regions in CRFR1cKO (mRNA levels presented are normalized to control levels, represented as dashed gray line, N = 3–6 per group, p = 0.373–0.991; data are presented as fold change). Image analysis of colocalization levels of Cre activity in the Δα6-Cre line and CRFR1 expression are presented in Figure 2-2. d–f, Cerebellar structural properties. No differences in cerebellar weight (N = 8, 7; p = 0.284; d) or cerebellar volume (quantification and representative image of MRI scan, N = 7, 8; p = 0.277; e) were detected. f, No differences in GrC soma size or number of dendrites were detected in CRFR1cKO compared with control mice (diameter: n = 43,51 cells, N = 2, p = 0.82; dendrites N = 3, 3; p = 0.45). Data are represented as mean ± SEM. ***p < 0.005.

Article Snippet: Lysates were subjected to gel electrophoresis on 10% SDS-PAGE and separated proteins were transferred onto nitrocellulose membranes and probed with AB5453 Rabbit Anti-GABA A Receptor α6 Antibody (Merck-Millipore; RRID: AB_177478 ) or Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783; Abcam; RRID: AB_303296 ).

Techniques: Expressing, Control, Activity Assay

Journal: The Journal of Neuroscience

Article Title: Cerebellar Learning Properties Are Modulated by the CRF Receptor

doi: 10.1523/JNEUROSCI.3106-15.2018

Figure Lengend Snippet: Whole cerebellum mRNA sequencing reveals significant differences in transcriptome of control and CRFR1cKO mice. a, Log2 mRNA analysis of CRFR1cKO and control cerebella mice revealed 563 significantly upregulated or downregulated genes (light purple dots; N = 6, 6). Ten genes showed log2 change larger than ±0.4 (fold change >30%, dark purple dots; 3 genes upregulated and 7 downregulated). b, Real-time qRT-PCR analysis was used to validate the downregulation of Atp2b4 mRNA levels in CRFR1cKO mice compared with controls (N = 7, 5; p < 0.001). c, Quantification by Western blot (WB) of PMCA4, the protein coded by the Atp2b4 gene, validates its translation downregulation in cerebella of CRFR1cKO (N = 7, 6; p = 0.014). Data are represented as mean ± SEM. *p < 0.05, &p < 0.001.

Article Snippet: Lysates were subjected to gel electrophoresis on 10% SDS-PAGE and separated proteins were transferred onto nitrocellulose membranes and probed with AB5453 Rabbit Anti-GABA A Receptor α6 Antibody (Merck-Millipore; RRID: AB_177478 ) or Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783; Abcam; RRID: AB_303296 ).

Techniques: Sequencing, Control, Quantitative RT-PCR, Western Blot

Journal: The Journal of Neuroscience

Article Title: Cerebellar Learning Properties Are Modulated by the CRF Receptor

doi: 10.1523/JNEUROSCI.3106-15.2018

Figure Lengend Snippet: CRFR1cKO mice do not show any motor phenotype under challenging or neutral conditions. a, b, Home cage locomotion measured in single-caged CRFR1cKO mice and control mice did not reveal any differences between the groups in light or dark phases (N = 7, 6; p = 0.607, 0.289, respectively). c, Mice tested on the rotarod for a single session (4 trials) at 10rpm/min or 20rpm/min (from 0 to 40 rpm in 4 or 2 min) lasted for similar latencies on the rod (10 rpm: N = 12, 21; p = 0.653; 20 rpm: N = 13, 9; p = 0.171). d, Layout of treadmill protocol. e, Mice tested on a treadmill fell onto the grid to a similar extent (N = 12,10, p = 0.849. f–h, CRFR1cKO and control mice do not show a fear- or anxiety-related phenotype. f, Mice do not exhibit differences in startle response (N = 13, 15; p = 0.242). g, CRFR1cKO and control mice tested in the open-field test do not show a significant difference in time spent in the center of the arena, number of visits to or distance in the center, or in total distance walked during the task (N = 14, 14; p = 0.11–0.9). h, After fear conditioning, CRFR1cKO and control mice did not show differences in freezing during cue (N = 6, 4; p = 0.332) or context (N = 6, 6; p = 0.289) presentation. Data are represented as mean ± SEM.

Article Snippet: Lysates were subjected to gel electrophoresis on 10% SDS-PAGE and separated proteins were transferred onto nitrocellulose membranes and probed with AB5453 Rabbit Anti-GABA A Receptor α6 Antibody (Merck-Millipore; RRID: AB_177478 ) or Anti-Calcium Pump PMCA4 ATPase antibody [JA9] (ab2783; Abcam; RRID: AB_303296 ).

Techniques: Control