Journal: Cell reports

Article Title: Retrograde adenosine/A 2A receptor signaling facilitates excitatory synaptic transmission and seizures

doi: 10.1016/j.celrep.2024.114382

Figure Lengend Snippet: (A) Left, Diagram illustrating the recording configuration. MC and MPP EPSCs were recorded from the same GC and evoked with stimulation electrodes placed in the inner and middle molecular layer, respectively. Right, Current clamp recording showing GC theta-burst firing (GC TBF). LTP induction protocol (GC TBF) was composed of 10 bursts at 5 Hz of 5 action potentials at 50 Hz, repeated 4 times every 5 s. (B) Left, Representative traces before (1) and after (2) GC TBF delivery. Right, Time-course plot showing that GC TBF induced LTP at MC-GC but not at MPP-GC synapses. (C) GC TBF-induced LTP was associated with significant reduction in PPR and CV (n = 13 cells). ** p < 0.01, *** p < 0.001. (D) LTP was abolished when TrkB was conditionally knocked out from from GCs (Post TrkB cKO, TrkBfl/fl mice injected in the dorsal blade with AAV5.CaMKII.Cre.GFP). LTP was unaffected in control animals (Control, TrkBfl/fl mice injected in the dorsal blade with AAV5.CaMKII.eGFP). (E) LTP was normally induced when loading PKI6-22 (2.5 μM) in GCs via the recording pipette but completely blocked when the cell-permeable PKA inhibitor PKI14-22 myristoylated (1 μM) was bath applied. (F) Summary bar graph showing the magnitude of GC TBF-induced LTP in the presence of DGC-IV (1 μM), when TrkB was conditionally knocked out from MCs (Pre TrkB cKO), when loading the PKI6-22 (2.5 μM) in GCs, and in the presence of D-APV (50 μM). LTP was abolished in the presence of the TrkB antagonist ANA-12 (15 μM), when Botox (0.5 μM) was loaded postsynaptically, in postsynaptic BDNF and TrkB cKO mice, and during bath application of the PKA inhibitors H89 (10 μM) or myristoylated PKI14-22 μM). Time-course summary plots are shown in Figure S1. ** p < 0.01, *** p < 0.001. (G) Scheme illustrating the emerging model for the mechanism underlying GC TBF-LTP. GC TBF triggers postsynaptic BDNF release and subsequent TrkB activation in GCs (1). Presynaptic PKA is then engaged downstream of postsynaptic BDNF/TrkB signaling (2), suggesting the requirement of a retrograde signal. Lastly, presynaptic PKA activation resulted in a long-lasting increase in glutamate release (3). Numbers in parentheses indicate the number of cells. Data are presented as mean ± SEM.

Article Snippet: H89 dihydrochloride , Tocris Bioscience , Cat#2910.

Techniques: Injection, Control, Transferring, Activation Assay

Journal: Cell reports

Article Title: Retrograde adenosine/A 2A receptor signaling facilitates excitatory synaptic transmission and seizures

doi: 10.1016/j.celrep.2024.114382

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: H89 dihydrochloride , Tocris Bioscience , Cat#2910.

Techniques: Virus, Plasmid Preparation, Recombinant, Software

Journal: Signal Transduction and Targeted Therapy

Article Title: Protein C receptor maintains cancer stem cell properties via activating lipid synthesis in nasopharyngeal carcinoma

doi: 10.1038/s41392-021-00866-z

Figure Lengend Snippet: Signaling cascade induced by activated PROCR in NPC. a Ca 2+ flux dynamics labeled by fluo-4 in PROCR overexpression or knockout cells. Living cells were observed right after the dye supplementation. The corresponding right panels show the statistical analysis of fluorescence intensity. b , c Western blotting analysis of lipid metabolism related genes expression and stem cell markers expression in vector or PROCR overexpressing cells with different drugs treatment. 8-Bromo-cAMP, PKA agonist; H 89 2HCl, PKA antagonist; PDTC, NFκB inhibitor. d ELISA detection of the cellular cAMP content in PROCR overexpressing cells. e Sphere forming assay of the sorted CD45 - EPCAM + PROCR + cells treated with different drugs. f , g The cell content of triglycerides and cholesterol in PROCR overexpressing cells treated with different drugs. h ChIP-PCR detection of the direct transcriptional regulation of NFκB on FASN or PTGS2 expression. i Subcutaneous xenograft tumor volume of PROCR overexpressing cells treated with different drugs; n = 5 for each group. All mice received APC activation. j Percentages of GFP + cells from xenograft tumor nodules in the lungs of mice treated with different drugs. The drugs were administered every week; ns Not significant, * P < 0.05, *** P < 0.001

Article Snippet: For cell culture, the drugs treatment dosages were as follows: APC (100 nM, 55-3-08 A, Pepmic, Suzhou, China), PKA agonist (8-Bromo-cAMP, 10 μM, S7857, Selleck, Houston, TX, USA), PKA antagonist (H 89 2HCl, 10 μM, S1582, Selleck), NFκB inhibitor PDTC (Pyrrolidinedithiocarbamate ammonium, 10 μM, S3633, Selleck), Ca 2+ -ATPase inhibitor (Thapsigargin, 1 μM, ab120286, Abcam), FASN inhibitor (C75, 50 μM, S8915, Selleck), PTGS1 and PTGS2 inhibitor (Ketoprofen, 50 μM, S1645, Selleck), cAMP agonist (Forskolin, 10 μM, S2449, Selleck) and calcium chelator (BAPTA-AM, 10 μM, HY-100545, MedChemExpress).

Techniques: Labeling, Over Expression, Knock-Out, Fluorescence, Western Blot, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Activation Assay