Journal: bioRxiv

Article Title: Dorsal peduncular cortex activity modulates affective behaviors in mice

doi: 10.1101/2023.04.25.538301

Figure Lengend Snippet: A-B. Experimental timeline. C57BL/6J mice were injected with AAV5-hSyn-mCherry, AAV5-hSyn-hM3D-mCherry or AAV5-hSyn-hM4D-mCherry into the DP/DTT. Following 2 weeks, mice underwent a battery of behavioral tests comprised of the open field test (OFT), elevated plus maze (EPM), tail suspension test (TST), forced swim test (FST) and auditory fear conditioning (Fear) with a washout period of three days between each test. C. Representative images of an mCherry control virus-infused brain stained for mCherry (red) and the nuclear dye Hoechst (blue). D-F. Diagrams depicting overlaid viral spread for all mCherry ( D ), hM4D ( E ) and hM3D-injected ( F ) mice. Scale bar = 500µm.

Article Snippet: The fear conditioning apparatus was located within a sound-attenuated chamber (63.5cm wide, 36.8cm, high, 74.9cm deep; catalog #NIR-022MD, Med Associates).

Techniques: Injection, Battery, Suspension, Control, Virus, Staining

Journal: bioRxiv

Article Title: Dorsal peduncular cortex activity modulates affective behaviors in mice

doi: 10.1101/2023.04.25.538301

Figure Lengend Snippet: A.-B. Experimental timeline. Mice previously injected with hM3D, hM4D or control mCherry constructs in the DP/DTT were injected with the DREADD agonist C21 one hour prior to undergoing auditory fear conditioning. During acquisition, mice were placed in a sound-attenuated fear conditioning chamber and received 6 tone-shock pairings (0.5mA). Twenty-four hours later, auditory fear memory was evaluated in a different context in the absence of shocks. B. During fear acquisition, DP/DTT manipulations did not affect average tone freezing. C. hM4D mice displayed significantly more freezing than hM3D mice at tone presentations 4 and 6 during acquisition. D-E. DP/DTT manipulations did not affect average or individual post shock freezing episodes. F-I. During fear retrieval, DP/DTT manipulations did not affect freezing during average tone ( F ), individual tone ( G ), average post tone ( H ) or individual post tone ( I ) periods. indicates a significant difference between hM3D and hM4D groups ( p <0.05). Male (square) and female (circle) individual datapoints are displayed for transparency (see text for details).

Article Snippet: The fear conditioning apparatus was located within a sound-attenuated chamber (63.5cm wide, 36.8cm, high, 74.9cm deep; catalog #NIR-022MD, Med Associates).

Techniques: Injection, Control, Construct

Journal: Communications Biology

Article Title: Membrane pools of phosphatidylinositol-4-phosphate regulate KCNQ1/KCNE1 membrane expression

doi: 10.1038/s42003-021-02909-1

Figure Lengend Snippet: a Schematic figure showing Pseudojanin (PJ), PJ-Sac, PJ-INPP5E, and PJ-dead constructs and the membrane targeted Lyn-11-FRB-mCherry construct. b Schematic figure showing Pseudojanin (PJ) and the membrane targeted Lyn-11-FRB-mCherry construct together with the PI4P biosensor SidM-GFP before and after rapamycin treatment. c Representative confocal images in HEK293T cells expressing PLCδ1-PH-GFP and the PJ constructs indicated (PJ, PJ-Sac, PJ-INPP5E or PJ-DEAD) before and after rapamycin treatment (1 μM, 30 min). d Summary data of PLCδ1-PH-GFP (top) and SidM-GFP (bottom) membrane to cytoplasmic fluorescence ratio (M/C) measured. Cells were treated as indicated. Scale bars, 5 μM. * p < 0.05. number of cells indicated in parenthesis.* p < 0.05.

Article Snippet: Additional constructs used were Pseudojanin (PJ), a fusion protein of Sac and INPP5E phosphatase domains with FKBP (Addgene, #37999); PJ-Sac, a PJ based construct containing an inactivating mutation (Asp1263Ala) in the INPP5E domain (Addgene, #38000); PJ-INPP5E, a PJ based construct with an inactivating mutation (Cys779Ser) in the SAC1 domain (Addgene, #38001); PJ-DEAD, a PJ based construct containing an inactivating mutation (Asp1263Ala) in the INPP5E domain and an inactivating mutation (Cys779Ser) in the SAC1 domain (Addgene, #38002) , ; PH-PLCδ1-GFP (Addgene, #51407); LYN11-FRB-mcherry (Addgene, #38004); GFP-P4M-SidM (Addgene, #51469), hKCNH2-EGFP (VectorBuilder); EGFP-Kir2.1 (Addgene, #107184); Kv7.4-EGFP (Addgene, #111453); Kv2.1-EGFP (Addgene, #111531); constitutively active PI4K (Addgene, #139311); dominant negative PI4K-dead (Addgene, #139312); KCNE1-KCNQ1 tandem .

Techniques: Construct, Membrane, Expressing, Fluorescence

Journal: Communications Biology

Article Title: Membrane pools of phosphatidylinositol-4-phosphate regulate KCNQ1/KCNE1 membrane expression

doi: 10.1038/s42003-021-02909-1

Figure Lengend Snippet: a Schematic figure showing Pseudojanin (PJ), PJ-Sac, PJ-INPP5E, and PJ-dead constructs and the membrane targeted Lyn-11-FRB-mCherry construct together with the channel before and after rapamycin treatment. b Left: representative HEK293T cells expressing KCNQ1-GFP, KCNE1, and the PJ constructs indicated after rapamycin treatment (1 μM, 30 min). Right: summary data of membrane to cytoplasmic fluorescence ratio (M/C) of KCNQ1-GFP measured in experiments as the left panel. Cells were treated as indicated. Scale bars, 5 µM. * p < 0.05, number of cells at least 27.

Article Snippet: Additional constructs used were Pseudojanin (PJ), a fusion protein of Sac and INPP5E phosphatase domains with FKBP (Addgene, #37999); PJ-Sac, a PJ based construct containing an inactivating mutation (Asp1263Ala) in the INPP5E domain (Addgene, #38000); PJ-INPP5E, a PJ based construct with an inactivating mutation (Cys779Ser) in the SAC1 domain (Addgene, #38001); PJ-DEAD, a PJ based construct containing an inactivating mutation (Asp1263Ala) in the INPP5E domain and an inactivating mutation (Cys779Ser) in the SAC1 domain (Addgene, #38002) , ; PH-PLCδ1-GFP (Addgene, #51407); LYN11-FRB-mcherry (Addgene, #38004); GFP-P4M-SidM (Addgene, #51469), hKCNH2-EGFP (VectorBuilder); EGFP-Kir2.1 (Addgene, #107184); Kv7.4-EGFP (Addgene, #111453); Kv2.1-EGFP (Addgene, #111531); constitutively active PI4K (Addgene, #139311); dominant negative PI4K-dead (Addgene, #139312); KCNE1-KCNQ1 tandem .

Techniques: Construct, Membrane, Expressing, Fluorescence