dpni  (Tocris)

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: Fig. 2. Symmetric attenuative propagation of GABAergic PSPs. (A and B) Left: Dendritic segments of interest used for voltage imaging of local GABA-evoked hyperpo- larizing (A) or depolarizing (B) postsynaptic potentials (PSPGABA). The magenta point or line indicates the locations of GABA uncaging. Middle: ΔF/F traces for PSPGABA at five ROIs [4 to 8 for (A) and 2 to 6 for (B) indicated in the left image]. Right: Relative ΔF/F for PSPGABA plotted against distance from the uncaged site [16 cells: red, 56; black, 59 regions for (A); 4 cells: red, 11; black, 12 regions for (B)]. Square plots indicate averages (±SEM). See also fig. S3.

Article Snippet: For glutamate or GABA uncaging, MNI- caged l- glutamate (300 μM, Tocris Bioscience) or DPNI- caged GABA (200 to 500 μM, Tocris Bioscience) was added to the external solution.

Techniques: Imaging

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: Fig. 3. [Cl−]in-dependent more negative membrane potential in dendrites. (A) An image of a direct gramicidin patch recording from a dendrite. Magnified view of the patched branch in yellow (left). (B) Vm trace recorded from a dendritic branch [shown in (A)]. (C) Vmrest 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) Vmrest 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) Vmrest (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 Vm indicated by different colors. (H) GABA currents at a dendrite (red) or at the soma (blue) plotted against the holding Vm. (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 inten- sity of GFP (green) and KCC2 (red) plotted against the distance from the soma.

Article Snippet: For glutamate or GABA uncaging, MNI- caged l- glutamate (300 μM, Tocris Bioscience) or DPNI- caged GABA (200 to 500 μM, Tocris Bioscience) was added to the external solution.

Techniques: Membrane, 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: Fig. 6. Validation of a model prediction: Critical role of Cl− conductance in EPSP amplification. (A and B) Left: Stimulated dendritic segment in the presence of NPPB (50 μM) and lidocaine [5 mM, (A)] or GABA [50 μM, (B)]. The magenta points indicate the locations of glutamate uncaging. Middle: ΔF/FeEPSP traces at five ROIs (1 to 5) in- dicated in the left image. Right: Relative ΔF/FeEPSP plotted against distance from the uncaged site (13 cells: red, 37; black, 34 regions for (A); 10 cells: red, 29; black, 31 re- gions for (B)]. Square plots indicate averages (±SEM). (C) Statistical summary of ΔF/FeEPSP amplification in the distal region with or without NPPB and lidocaine, and GABA (**P < 0.01 and ***P < 0.001, Dunnett’s test). Error bars are the SEM.

Article Snippet: For glutamate or GABA uncaging, MNI- caged l- glutamate (300 μM, Tocris Bioscience) or DPNI- caged GABA (200 to 500 μM, Tocris Bioscience) was added to the external solution.

Techniques: Biomarker Discovery, Amplification

Journal: Cell Reports

Article Title: KCC2-Mediated Cl − Extrusion Modulates Spontaneous Hippocampal Network Events in Perinatal Rats and Mice

doi: 10.1016/j.celrep.2019.01.011

Figure Lengend Snippet: Inhibition of KCC2 Increases GABA-Driven Spontaneous Spiking in CA3 Pyramidal Neurons, but Not in Interneurons (A) KCC2 expression in E18.5 GAD67-GFP mouse hippocampal CA3 area. The areas indicated with rectangles (left) are shown in more detail on the right. Examples of KCC2-expressing interneurons and pyramidal neurons are indicated with arrowheads and stars, respectively. (B) Scheme of experimental paradigm to record interneuronal activity from a pyramidal neuron (left). Sample traces of a whole-cell voltage-clamp recording (middle) and median of the normalized frequency values (right) of sIPSCs recorded from pyramidal neurons before and after the application of VU0463271 (10 μM) are shown. (C) Scheme of experimental paradigm to record pyramidal neuron activity in loose cell-attached configuration (left). Sample traces of a loose cell-attached recording of CA3 pyramidal neuron spiking (middle) and median of the normalized spike frequency (right) in the presence and absence of VU0463271 and picrotoxin (100 μM) are shown. Recordings in (B) and (C) were done in the presence of CNQX (10 μM) and d -AP5 (20 μM), indicated in the figure as iGluR block. Data are presented as median and IQR; Wilcoxon signed-rank test was used for statistical analysis; ∗ p < 0.05; ∗∗ p < 0.01; n values are indicated in the bar diagrams.

Article Snippet: DPNI-gaged GABA , Tocris , Cat #: 2991.

Techniques: Inhibition, Expressing, Activity Assay, Blocking Assay

Journal: Cell Reports

Article Title: KCC2-Mediated Cl − Extrusion Modulates Spontaneous Hippocampal Network Events in Perinatal Rats and Mice

doi: 10.1016/j.celrep.2019.01.011

Figure Lengend Snippet: VU0463271-Sensitive Chloride Extrusion in Neonatal CA3 Pyramidal Neurons (A) An Alexa Fluor 488-filled CA3 pyramidal neuron from a P1 rat representing the position of the whole-cell patch pipette imposing a 19 mM Cl − load onto the neuron and the sites of UV photolysis of caged GABA. (B) Chloride extrusion efficacy in P1 rat CA3 pyramidal neurons determined using local UV photolysis of caged GABA at the soma (n = 4) and apical dendrite at 50 μm (n = 4) and 200 μm (n = 5) distance from the somatic Cl − load, under control conditions (black) and in the presence of VU0463271 (10 μM, gray). Stippled line delimits the approximate residual negative deflection in E GABA from E GABA-GHK at 50 μm that is insensitive to VU0463271 (see Spoljaric et al., 2017 ) and present in KCC2 −/− neurons (see Li et al., 2007 ). (C) Sample E GABA recordings and their corresponding I-V plots in the presence of iGluR block and bumetanide (2.5 μM). (D) Sample traces of gramicidin-perforated patch recordings (left) and median of the normalized amplitude of miniature inhibitory postsynaptic currents (mIPSCs; right) recorded from pyramidal neurons before and after the application of VU0463271 or DMSO. Recordings were done in the presence of iGluR block and TTX (0.5 μM). Data in (B) and (D) are presented as median and IQR; Wilcoxon signed-rank test were used for statistical analysis; ∗ p < 0.05; n values are indicated in the bar diagram.

Article Snippet: DPNI-gaged GABA , Tocris , Cat #: 2991.

Techniques: Transferring, Control, Blocking Assay

Journal: Cell Reports

Article Title: KCC2-Mediated Cl − Extrusion Modulates Spontaneous Hippocampal Network Events in Perinatal Rats and Mice

doi: 10.1016/j.celrep.2019.01.011

Figure Lengend Snippet:

Article Snippet: DPNI-gaged GABA , Tocris , Cat #: 2991.

Techniques: Recombinant, Software