Journal: The Journal of Biological Chemistry

Article Title: An Antibody Biosensor Establishes the Activation of the M 1 Muscarinic Acetylcholine Receptor during Learning and Memory * ^♦

doi: 10.1074/jbc.M115.681726

Figure Lengend Snippet: M 1 mAChR is activated in the hippocampus following drug treatment and memory acquisition. A, C57/BL6/NTAC mice were injected (i.p.) with vehicle or xanomeline (5 mg/kg). After 30 min, tissues were fixed by transcardial perfusion, and sections were obtained and stained with the phospho-specific serine 228 antibody and with DAPI stain to reveal the nuclei. Shown are representative sections through the CA1 region of the hippocampus. B, C57/BL6/NTAC mice ( Wild-Type ) or M 1 mAChR-knock-out mice ( M1-KO ) were injected (intraperitoneally) with BQCA (15 mg/kg) or vehicle, and after 30 min hippocampal membranes were prepared from which the M 1 mAChR was immunoprecipitated. The sample was then processed in Western blots, which were probed with phospho-specific serine 228 antibody or an M 1 mAChR-specific antibody to detect total M 1 mAChR. C, quantification of Western blots from B . The data are presented as means ± S.E. ( n = 3). Statistical analysis uses Student's paired t test. D, C57/BL6/NTAC mice were subjected to a fear conditioning training protocol or to an unpaired immediate foot shock as a control; 30 min later tissue was fixed by transcardial perfusion, and sections obtained and stained with phosphorylated Ser 228 -specific antibody ( upper panel ) or anti-c-FOS antibody ( lower panel ). All the data shown are typical of at least three independent experiments.

Article Snippet: Male C57Bl6/NTAC mice (8–15 weeks old) were placed in the conditioning chamber (Stoelting ANY-maze fear conditioning system), and after a 2-min adaptation period, the mice received three tone/foot shock pairings, where a tone (conditioned stimulus, 2.8 kH, 85 db, 30 s) always co-terminated with the foot shock (unconditioned stimulus, 2 s, 0.4 mA).

Techniques: Injection, Staining, Knock-Out, Immunoprecipitation, Western Blot, Control

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Phosphodiesterase Inhibition Rescues Chronic Cognitive Deficits Induced by Traumatic Brain Injury

doi: 10.1523/JNEUROSCI.5133-12.2013

Figure Lengend Snippet: Cue and contextual conditioning at 2 weeks after TBI or sham surgery (n = 8/group). Animals received vehicle (5% DMSO in saline) or rolipram (0.03 mg/kg) 30 min before training. Fear conditioning was assessed at 24 h (A) or 4 weeks (B) after training without rolipram treatment. Both cue and contextual freezing were significantly increased in TBI animals treated with rolipram compared with vehicle-treated TBI animals. No significant differences between groups were observed in baseline freezing on the training day (Training) or baseline freezing on the testing day before the cue in the new context (Baseline). *p < 0.05, **p < 0.01, ***p < 0.001 (two-way ANOVA with Bonferroni’s post hoc comparison).

Article Snippet: The fear conditioning apparatus (30.5 × 24.1 × 21 cm, Coulbourn Instruments) had an electric grid floor (1.6 cm spacing, 4.8 mm diameter rods).

Techniques: Saline, Comparison

Journal: The Journal of neuroscience : the official journal of the Society for Neuroscience

Article Title: Phosphodiesterase Inhibition Rescues Chronic Cognitive Deficits Induced by Traumatic Brain Injury

doi: 10.1523/JNEUROSCI.5133-12.2013

Figure Lengend Snippet: Phosphorylated CREB levels were assessed in the hippocampus at 20 min after cue and contextual fear conditioning. A, Representative Western blots. B, Densitometric analysis. CREB phosphorylation significantly increased in sham animals after training, but not in TBI animals treated with vehicle 30 min before training (n = 6/group). Rolipram rescued the TBI-induced deficits in CREB phosphorylation after learning. *p < 0.05 versus untrained sham + vehicle animals, **p < 0.01 versus untrained sham + vehicle animals, (one-way ANOVA with Bonferroni’s post hoc comparison). #p < 0.01 for TBI + vehicle versus TBI + rolipram animals (one-way ANOVA with Bonferroni’s post hoc comparison).

Article Snippet: The fear conditioning apparatus (30.5 × 24.1 × 21 cm, Coulbourn Instruments) had an electric grid floor (1.6 cm spacing, 4.8 mm diameter rods).

Techniques: Western Blot, Phospho-proteomics, Comparison

Journal: PLoS ONE

Article Title: Therapeutic benefits of phosphodiesterase 4B inhibition after traumatic brain injury

doi: 10.1371/journal.pone.0178013

Figure Lengend Snippet: (A) At 24 hrs and 1 month after training, animals were evaluated for contextual fear conditioning. At both 24 hrs and 1 month after training, vehicle-treated TBI animals froze significantly less than sham animals. A33-treated TBI animals froze significantly more than vehicle-treated TBI at 24 hrs, but not at 1 month, after training. A main effect of treatment ( F (2, 72) = 6.996, p = 0.003), a main effect of trial ( F (2, 72) = 169.111, p <0.001) and a significant interaction of treatment x trial ( F (4, 72) = 4.931, p = 0.001) were observed. *p <0.05, ***p <0.001 vs. Sham Day 1, **p <0.01 vs. Sham Day 30, # p <0.001 vs. Training, + p <0.01 vs. TBI+Vehicle Day 1, repeated-measures two-way ANOVA with post-hoc Student-Newman-Keuls. (B) Shock threshold was similar between all treatment groups. Mean ± SEM, n = 12-14/group, one-way ANOVA with post-hoc Student-Newman-Keuls.

Article Snippet: Animals were habituated to the fear conditioning apparatus (30.5 x 24.1 x 21 cm, Coulbourn Instruments) for 10 min to facilitate contextual fear conditioning [ ].

Techniques: