Journal: Journal of Neuroscience

Article Title: Pro-Brain-Derived Neurotrophic Factor Inhibits GABAergic Neurotransmission by Activating Endocytosis and Repression of GABAA Receptors

doi: 10.1523/jneurosci.2069-14.2014

Figure Lengend Snippet: Figure6. ProBDNFhasnoeffectonphospho-CREBactivationbutinducesICERexpressionviatheJAK2/STAT3signalingcascade.A,ImmunofluorescencesignalofCREB(red,top),pCREB(green, middle)inhippocampalneuronsincontrolconditions(CTR),ortreatedwithCR-proBDNFalone(25ng;CR-proBDNF),orCR-proBDNFplusmBDNF(CR-proBDNF mBDNF;25ng),orCR-proBDNFplus mBDNF in the presence of K252a (CR-proBDNF mBDNF K252a; 200 nM). Right, Merged images with MAP2 staining (blue). Scale bar, 40 m. B, Average ratio of pCREB/CREB in the different conditionsnormalizedtocontrol.n15neuronsanalyzedpertreatmentinn3independentexperiments;***p0.001.C,ImmunofluorescencesignalofICER(red;top)inhippocampalneurons in control conditions (CTR), or treated with CR-proBDNF (25 ng) for 30 min (CR-proBDNF 30 min), for 12 h (CR-proBDNF 12 h), or for 30 min in the presence of TAT-pep5 (CR-pro-BDNF 30 min TATpep5;2M),followingacostainingwithanti-MAP2chickenpolyclonalantibody(green;bottom)torevealneuronalcells.Scalebar:20m.D,AveragefluorescenceintensityratioofICER/MAP2 in the different conditions normalized to control. Forty-five neurons analyzed per condition, n 4; ***p 0.001. E, Western blot analysis with the antibody to ICER. ICER expression was upregulatedafteracutetreatmentwithCR-proBDNF(25ng,30min)butnotwithchronicapplicationofCR-proBDNF(25ng,12h).TheincreaseofICERwasantagonizedbyTAT-pep5(2M),AG490 (50 M), and Stattic (10 M). This increase was insensitive to ROCKi (50 M). Proteins were visualized using ECL following incubation with HRP-conjugated secondary (Figure legend continues.)

Article Snippet: TTX was purchased from Abcam; ROCKi (Y-27632) was purchased from Millipore; tPA-Stop was purchased from American Diagnostica; k252a, TAT-pep5, and Stattic were purchased from Calbiochem; nifedipine, aprotinin, leupeptin, and AG490 were purchased from Sigma-Aldrich; CR-proBDNF recombinant (Escherichia coli) mutated human and mBDNF were purchased from Alomone Labs. NBQX, CNQX, and D-APV were obtained from the Molecular, Cellular, and Genomic Neuroscience Research Branch of the National Institute of Mental Health.

Techniques: Staining, Control, Western Blot, Expressing, Incubation

Journal: Journal of Neuroscience

Article Title: Programmed and Induced Phenotype of the Hippocampal Granule Cells

doi: 10.1523/jneurosci.1674-05.2005

Figure Lengend Snippet: Figure 1. Mossy fiber monosynaptic GABAergic transmission can be detected when GCs express the markers of the GABAergic phenotype. A, Synaptic responses recorded in CA3 pyra- midalcellsfromnaiveslicesareblockedbytheperfusionoftheionotropicglutamatereceptors antagonists NBQX and APV. B, In contrast, in preparations exposed to 30 min of BDNF (or to synaptic or direct activation in vivo and in vitro), mossy fiber stimulation provokes bicuculline- sensitive monosynaptic IPSPs in the presence of NBQX and APV. C, BDNF treatment does not induce hyperexcitability, as assessed by field potential responses of CA3 to DG stimulation. D, The Trk receptor inhibitor K252a prevents the effect of BNDF, described in B. E, The pharmaco- logicallyisolatedmonosynapticGABAergicresponsesaredepressedbyL-AP-4,agroupIIImGluR agonist. All traces are an average of six evoked responses. BIC, Bicuculline; LAP-4, L-AP-4.

Article Snippet: The drugs used were diluted in the ACSF, namely, the NMDA receptor antagonist APV (30 M; Tocris Cookson), the nonNMDA receptor antagonist NBQX (10 M; Tocris Cookson), the GABAA receptor antagonist bicuculline methiodide (20 M; Sigma), and the group III metabotropic GluR (mGluR) agonist L( )-2-amino-4phosphonobutyric acid (L-AP-4) (1 M; Tocris Cookson).

Techniques: Transmission Assay, Activation Assay, In Vivo, In Vitro

Journal: Stem cell research

Article Title: Characterization and application of electrically active neuronal networks established from human induced pluripotent stem cell-derived neural progenitor cells for neurotoxicity evaluation.

doi: 10.1016/j.scr.2020.101761

Figure Lengend Snippet: Fig. 5. Modifications of electrical activity by acute pharmacological treatment. Ratio of treatment/control of CINDA-NN and rNN treated with GABA [1 mM], bicuculline [10 μM], glutamate [100 µM], AP5 [20 µM] and NBQX [10 µM]. Results are depicted as A) MFR, B) MBR and C) Spikes/Burst as mean+SEM. hiNPC: n = 7–10, rNN: n = 6–10. D and E Representative 60 s SRP of hNN and rNN of baseline- and GABA (D) or bicuculline treatment (E). *Significant within condition (p < 0.05).

Article Snippet: NBQX disodium salt, DL-AP5 sodium salt and Bicuculline were obtained from Santa Cruz Biotechnologies (Texas, USA).

Techniques: Activity Assay, Control

Journal: International Journal of Molecular Sciences

Article Title: Role of L-Type Voltage-Gated Calcium Channels in Epileptiform Activity of Neurons

doi: 10.3390/ijms221910342

Figure Lengend Snippet: The effects of different VGCC blockers on KCl-induced Ca 2+ response in neurons. ( A , B , D ) Effects of different doses of verapamil, nifedipine, and ML-218 (T-type VGCC blocker) on the amplitude of KCl-induced (35 mM) Ca 2+ response in the presence of NMDAR (D-AP5, 10 μM) and AMPAR/KAR (NBQX, 10 μM) antagonists. The pauses between KCl applications were 15 min. The traces of some representative neurons are shown in each panel. N = 100, n = 4 for each experiment. ( C , D′ ) Diagrams showing dose-dependent changes in the mean amplitude ratio in the presence of the blocker to mean amplitude in control. One-way ANOVA followed by Tukey’s multiple comparisons test. Insignificant changes are marked as n/s; p < 0.05 (*), p < 0.01 (**), p < 0.001 (***). ( E , E′ ) Traces of neurons ( E ) and diagram ( E′ ) demonstrating changes in the ratio of mean amplitude in control to mean amplitude in the presence of nifedipine, ML-218, ω-conotoxin MVIIC (blocker of P/Q- and N-type VGCC). N = 100, n = 4. One-way ANOVA followed by Tukey’s multiple comparisons test. p < 0.01 (**).

Article Snippet: The reagents that were used in experiments are listed below: Paraformaldehyde (P6148), Poly(ethyleneimine) solution (P3143), penicillin–streptomycin (P4333), (+)-cis-Diltiazem hydrochloride (D2521), (±)-Verapamil hydrochloride (V4629), Isradipine (I6658), Nifedipine (N7634) (Sigma-Aldrich, Saint Louis, MO, USA), Neurobasal-A medium (10888022), B-27 supplement (17504044), Trypsin 2.5% (15090046), Goat serum New Zeland origin (16210072) (Life Technologies, Grand Island, NY, USA), Fura-2 AM (F1221), Hoechst 33,342 Trihydrochloride Trihydrate (H1399) (Molecular Probes, Eugene, OR, USA), ML 218 hydrochloride (4507) (Tocris Bioscience, Bristol, UK), NBQX disodium salt (N-186), D-AP5 (D-145) (Alomone Labs, Jerusalem, Israel), Bicuculline (11727) (Cayman Chemical, Ann Arbor, MI, USA), goat anti-mouse Alexa Fluor 647 antibody (ab150115) (Abcam, Cambridge, UK), monoclonal mouse anti-GFAP antibodies (GF1 clone, L18/03) (Bialexa, Moscow, Russian Federation); Triton X-100 (Am-O694) (Amresco LLC, Solon, OH, USA); EGTA (A-0878), EDTA (A5097), (AppliChem, Darmstadt, Germany); HEPES (Cat. No 3350) (Dia-M, Moscow, Russian Federation).

Techniques: