Journal: Science Advances

Article Title: Cl − -dependent amplification of excitatory synaptic potentials at distal dendrites revealed by voltage imaging

doi: 10.1126/sciadv.adj2547

Figure Lengend Snippet: ( A ) An image of a direct gramicidin patch recording from a dendrite. Magnified view of the patched branch in yellow (left). ( B ) V m trace recorded from a dendritic branch [shown in (A)]. ( C ) V m rest measured by gramicidin patch from dendrites ( n = 8 cells) and soma ( n = 10 cells) is plotted against the distance from the soma. ( D ) Somatic (blue, n = 19 cells) and dendritic (green, n = 11 cells) V m rest with or without 10 μM VU0463271 (soma: yellow, n = 9 cells; dendrite: red, n = 11 cells). Dunnett’s test, ** P < 0.01; n.s., not significant. ( E and F ) V m rest (E) and leak conductance (F) plotted against distance from the soma. VU, VU0463271. ( G ) Representative gramicidin patch currents recorded from the soma (left) or a dendritic branch (right) in response to spot uncaging of DPNI-caged GABA at the recording sites, with altered holding V m indicated by different colors. ( H ) GABA currents at a dendrite (red) or at the soma (blue) plotted against the holding V m . ( I ) Images of immunostained KCC2 (magenta) in green fluorescent protein (GFP)–labeled hippocampal neurons (green). Three rectangular areas (a to c) were expanded with merged colors (middle), and the fluorescent signals for KCC2 and GFP along a line in each area are plotted on the right. ( J ) Relative fluorescent intensity of GFP (green) and KCC2 (red) plotted against the distance from the soma.

Article Snippet: In some experiments, TTX (1 μM, FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan), lidocaine (5 mM, Nacalai Tesque), D-AP5 (100 μM, Tocris Bioscience, Bio-Techne SRL, Minneapolis, MN, USA), Cd 2+ (100 μM, FUJIFILM Wako Pure Chemical Corporation), ZD7288 (100 μM, Tocris bioscience), VU0463271 (10 μM, Tocris Bioscience), and NPPB (50 μM, Tocris Bioscience) were used to inhibit Na + channels, NMDARs, Ca 2+ channels, HCN channels, KCC2, and Cl − channels, respectively.

Techniques: Labeling

Journal: Science Advances

Article Title: Cl − -dependent amplification of excitatory synaptic potentials at distal dendrites revealed by voltage imaging

doi: 10.1126/sciadv.adj2547

Figure Lengend Snippet: ( A to C ) Left: Images for dendritic segment of interest (ROIs 1 to 5) and Δ F / F eEPSP traces measured in the presence of VU0463271 [10 μM, (A)], lidocaine [5 mM, (B)], or both (C). The magenta points indicate the locations of glutamate uncaging. Middle: Relative Δ F / F eEPSP plotted against the distance from the uncaged site [17 cells: red, 44; black, 52 regions for (A); 17 cells: red, 55; black, 56 regions for (B); 15 cells: red, 38; black, 44 regions for (C)]. Square plots indicate averages (±SEM). Right: Absolute Δ F / F eEPSP sizes in the distal region of dendrites and uncaged sites [(A), n = 17 cells; (B), n = 17 cells; (C), n = 15 cells]. Paired t test. Square plots indicate averages (±SEM). ( D ) Statistical summary of Δ F / F eEPSP amplification in distal branches with or without VU0463271 (10 μM), lidocaine (5 mM), tetrodotoxin (TTX; 1 μM), Cd 2+ (100 μM), D-AP5 (100 μM), and ZD7288 (100 μM). Dunnett’s test, * P < 0.05. Error bars are the SEM. See also fig. S4.

Article Snippet: In some experiments, TTX (1 μM, FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan), lidocaine (5 mM, Nacalai Tesque), D-AP5 (100 μM, Tocris Bioscience, Bio-Techne SRL, Minneapolis, MN, USA), Cd 2+ (100 μM, FUJIFILM Wako Pure Chemical Corporation), ZD7288 (100 μM, Tocris bioscience), VU0463271 (10 μM, Tocris Bioscience), and NPPB (50 μM, Tocris Bioscience) were used to inhibit Na + channels, NMDARs, Ca 2+ channels, HCN channels, KCC2, and Cl − channels, respectively.

Techniques: Amplification

Journal: Science Advances

Article Title: Cl − -dependent amplification of excitatory synaptic potentials at distal dendrites revealed by voltage imaging

doi: 10.1126/sciadv.adj2547

Figure Lengend Snippet: ( A ) Left: Image of an ASAP3β-expressing neuron, with a yellow rectangular area expanded in the right, showing 10 sites of local glutamate uncaging (magenta, 35-ms intervals) in three patterns: to distal (a to j), to soma (j to a), or random. ( B ) Averaged Δ F / F eEPSP traces (±SEM, 17 cells) at three ROIs (uncaged branch: black or gray; proximal: blue; distal: red or orange as indicated in (A), right] upon three patterns of glutamate uncagings. The magenta lines indicate spot laser illumination. ( C ) Δ F / F eEPSP traces (means ± SEM) at ROI distal in three conditions: control (red or orange, 17 cells), with lidocaine (lido; 5 mM) and VU0463271 (10 μM) (purple, 13 cells), and with Cd 2+ (100 μM) and D-AP5 (100 μM) (yellow, 14 cells). ( D ) Comparison of synaptic integration at ROI distal upon two patterns of glutamate inputs in the absence (ctrl, top) or presence of lidocaine and VU0463271 (bottom). Two-way ANOVA, ** P < 0.01. ( E ) Ratio of Δ F / F eEPSP signals (means ± SEM) upon opposite directions of glutamate uncagings (“to soma” relative to “to distal”) at ROI distal in the absence (ctrl) or presence of lidocaine and VU0463271. Two-way ANOVA, ** P < 0.01. See also fig. S6.

Article Snippet: In some experiments, TTX (1 μM, FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan), lidocaine (5 mM, Nacalai Tesque), D-AP5 (100 μM, Tocris Bioscience, Bio-Techne SRL, Minneapolis, MN, USA), Cd 2+ (100 μM, FUJIFILM Wako Pure Chemical Corporation), ZD7288 (100 μM, Tocris bioscience), VU0463271 (10 μM, Tocris Bioscience), and NPPB (50 μM, Tocris Bioscience) were used to inhibit Na + channels, NMDARs, Ca 2+ channels, HCN channels, KCC2, and Cl − channels, respectively.

Techniques: Expressing, Control, Comparison

Journal: Science Advances

Article Title: Cl − -dependent amplification of excitatory synaptic potentials at distal dendrites revealed by voltage imaging

doi: 10.1126/sciadv.adj2547

Figure Lengend Snippet: ( A and B ) Left: Images of jGCaMP7f-expressing neuronal distal dendrites, the middle of which received local glutamate uncaging (magenta) five times at 50 Hz with different laser energies (0.65, 1.2, 1.8, 2.3, or 2.9 μJ) in the absence [control, (A)] or presence of lidocaine and VU0463271 (B). Right: Δ F / F of jGCaMP7f at six ROIs (indicated in the left image) upon 405-nm laser illumination at 1.2 or 2.9 μJ. ( C ) Δ F / F of jGCaMP7f (means ± SEM) in response to five different powers of laser illumination with (right) or without lidocaine and VU0463271 (left) is plotted against the distance from the uncaged site. n = 9 (control) and 15 cells (lidocaine and VU0463271).

Article Snippet: In some experiments, TTX (1 μM, FUJIFILM Wako Pure Chemical Corporation, Tokyo, Japan), lidocaine (5 mM, Nacalai Tesque), D-AP5 (100 μM, Tocris Bioscience, Bio-Techne SRL, Minneapolis, MN, USA), Cd 2+ (100 μM, FUJIFILM Wako Pure Chemical Corporation), ZD7288 (100 μM, Tocris bioscience), VU0463271 (10 μM, Tocris Bioscience), and NPPB (50 μM, Tocris Bioscience) were used to inhibit Na + channels, NMDARs, Ca 2+ channels, HCN channels, KCC2, and Cl − channels, respectively.

Techniques: Expressing, Control

Journal: The Journal of Physiology

Article Title: Ionotropic and metabotropic kainate receptor signalling regulates Cl − homeostasis and GABAergic inhibition

doi: 10.1113/JP276901

Figure Lengend Snippet: Kainate receptors activated by 1 μm KA modulate IPSCs in CA3 pyramidal cells in the presence of zinc chelator ZX1

Article Snippet: The sources of the agonists and antagonists used in the experiments were: KA (Sigma‐Aldrich, St Louis, MO, USA), NEM (Sigma‐Aldrich), GDP‐β‐S (Sigma‐Aldrich), dl ‐APV (Tocris Bioscience, St Louis, MO, USA), GYKI 52466 (Tocris Bioscience), {"type":"entrez-protein","attrs":{"text":"CGP55845","term_id":"875097176","term_text":"CGP55845"}} CGP55845 (Tocris Bioscience), dl ‐AP3 (Tocris Bioscience), ZX1 (Strem Chemicals, Newbury Port, MA, USA) and VU 0463271 (Tocris Bioscience).

Techniques:

Journal: The Journal of Physiology

Article Title: Ionotropic and metabotropic kainate receptor signalling regulates Cl − homeostasis and GABAergic inhibition

doi: 10.1113/JP276901

Figure Lengend Snippet: Kainate receptors activated by 0.1 μm KA modulate IPSCs in CA3 pyramidal cells in the presence of zinc chelator ZX1

Article Snippet: The sources of the agonists and antagonists used in the experiments were: KA (Sigma‐Aldrich, St Louis, MO, USA), NEM (Sigma‐Aldrich), GDP‐β‐S (Sigma‐Aldrich), dl ‐APV (Tocris Bioscience, St Louis, MO, USA), GYKI 52466 (Tocris Bioscience), {"type":"entrez-protein","attrs":{"text":"CGP55845","term_id":"875097176","term_text":"CGP55845"}} CGP55845 (Tocris Bioscience), dl ‐AP3 (Tocris Bioscience), ZX1 (Strem Chemicals, Newbury Port, MA, USA) and VU 0463271 (Tocris Bioscience).

Techniques: