Journal: Antioxidants (Basel, Switzerland)

Article Title: A Novel Gastrodin Derivative with Neuroprotection Promotes NGF-Mimic Activity by Targeting INSR and ACTN4 to Activate PI3K/Akt Signaling Pathway in PC12 Cells.

doi: 10.3390/antiox14030344

Figure Lengend Snippet: Figure 3. Screening NGF-mimic activity of GAD037-related potential targets and signaling pathways. (A–I) Effects of inhibitors of TrkA, TrkB, Ras, Raf, ERK, GR, IGF-1R, INSR, PI3K (K252a, ANA-12, FTA, AZ628, U0126, RU486, T9576, HNMPA-(AM)3, and LY294002, respectively) on neurite outgrowth in- duced by GAD037 treatment for 48 h. All experiments with inhibitors were conducted simultaneously in 3rd, 4th, and 9th generations of cells. Data are presented as mean ± SEM. *** p < 0.001 represents significant differences compared with control group; ns indicate no significant differences; # and ### indicate significant differences at p < 0.05 and p < 0.001, respectively.

Article Snippet: The pronase E (CAT No.: HY-114158) was bought from MedChemExpress, Shanghai, China, while tyrosine kinase B (TrkB) inhibitor (ANA-12, CAT No.: S7745) was acquired from Selleck, Shanghai, China.

Techniques: Activity Assay, Protein-Protein interactions, Control

Journal: Nature Communications

Article Title: BDNF rescues BAF53b-dependent synaptic plasticity and cocaine-associated memory in the nucleus accumbens

doi: 10.1038/ncomms11725

Figure Lengend Snippet: ( a ) Bath application of 500 nM ANA12 blocked theta burst-induced LTP (TBS 1 ) in NAc ( n =6) in slices from WT mice. A 60-min period of ANA12 washout restored LTP after a second bout of theta stimulation (TBS 2 ) in the same stimulated pathway. Inset, representative traces collected during baseline (grey line), 60 min post-TBS 1 (blue line) and 60 min post-TBS 2 (dark blue line). Scale bar, 0.4 mV, 5 ms. ( b ) Infusion of BDNF (2 nM) began after a 15-min stable baseline recording. BDNF produced a rapid increase in glutamatergic transmission (∼35%) within the NAc in slices from both WT ( n =6) and Baf53bΔHD ( n =5) mice. Before delivery of theta bursts, stimulation intensity was reduced to reset baseline to pre-drug levels, to evaluate the effects of BDNF on the magnitude of LTP (dotted downward arrow). Five theta trains (five theta bursts each, separated by 1 min) delivered 5 min after re-establishing stable baseline recordings (upward arrow) produced robust and stable LTP in WT slices and completely reversed the LTP impairment previously seen in slices from Baf53bΔHD mice (see ). Inset, representative traces collected during baseline (grey line), 30 min after start of BDNF infusion (blue line) and 60 min post TBS (dark blue line). Scale bar. 0.4 mV, 5 ms. ( c ) Pretreatment with 500 nM ANA12 completely blocked BDNF-induced increases in baseline synaptic transmission. The delivery of TBS produced a brief short-term potentiation that rapidly returned towards baseline levels ( n =6). Inset, representative traces collected during baseline (grey line) and 60 min post TBS (black line). Scale bar, 0.5 mV, 5 ms.

Article Snippet: The AMPA receptor antagonist, DNQX (Tocris) and human recombinant form of BDNF (Chemicon) were prepared fresh in aCSF, whereas the TrkB receptor antagonist, ANA12 (Tocris), stock was dissolved in dimethyl sulfoxide and then diluted to working concentration in aCSF (dimethyl sulfoxide <0.01%).

Techniques: Produced, Transmission Assay

Journal: Scientific Reports

Article Title: Blockade of soluble epoxide hydrolase attenuates post-ischemic neuronal hyperexcitation and confers resilience against stroke with TrkB activation

doi: 10.1038/s41598-017-18558-6

Figure Lengend Snippet: The functional relevance of TrkB overactivation in the sEH knockout (sEH KO) mice evaluated by pretreatment with the selective TrkB antagonist ANA12 before oxygen-glucose deprivation (OGD) ex vivo and before middle cerebral artery occlusion (MCAO) in vivo . ( A ) Representative traces of the field excitatory postsynaptic potentials (fEPSPs) in the hippocampus at different time points at baseline, during OGD induction and during post-ischemic long-term potentiation (iLTP) expression. The averaged time course of fEPSP changes over 60 minutes following OGD induction is shown for WT and sEH KO mice with either vehicle or ANA12 pretreatment. ( B ) During iLTP expression in the last 10 minutes, the average fEPSP amplitude significantly decreased in the sEH KO/vehicle group relative to the WT/vehicle group; this effect was abolished by ANA12 pretreatment in the sEH KO group (sEH KO/ANA12, n = 6/group). ( C ) Vital staining of representative brain slices at 48 hours after MCAO from WT and sEH KO mice with either vehicle or ANA12 pretreatment. ( D ) ANA12 pretreatment (sEH KO/ANA12, n = 6) significantly eliminated the infarct reduction in the sEH KO/vehicle relative to the WT/vehicle (n = 6/group), suggesting that the protective effects of sEH deletion on the attenuation of iLTP and infarction were mediated by TrkB activation. ( E ) A mechanistic framework summarizing the effects of sEH blockade. Blocking sEH induces TrkB overactivation and prevents EET degradation, leading to neuroprotection, iLTP reduction, vasodilatation, and, finally, reductions in infarcts and their associated deficits. Calibration: 0.2 mV, 5 ms. The values represent the mean ± standard error of the mean. *, # p < 0.05, ***, ### p < 0.001.

Article Snippet: WT and sEH KO mice were pretreated with intraperitoneal injection of ANA12 (0.5 mg/kg, MedKoo; n = 5/group) or vehicle (1% DMSO, Sigma-Aldrich; n = 3/group) every 12 hours for 3 times before the MCAO surgery to ensure TrkB inhibition.

Techniques: Functional Assay, Knock-Out, Ex Vivo, In Vivo, Expressing, Staining, Activation Assay, Blocking Assay