qx 314  (Alomone Labs)


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    Structured Review

    Alomone Labs qx 314
    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including <t>QX-314</t> in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Qx 314, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    2) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    3) Product Images from "Generation of rhythmic patterns of activity by ventral interneurones in rat organotypic spinal slice culture"

    Article Title: Generation of rhythmic patterns of activity by ventral interneurones in rat organotypic spinal slice culture

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.1999.0459t.x

    Rhythmic activity induced by high-K + solution under current clamp recording A , rhythmic episodes evoked by 7 mM K + recorded at three different levels of resting membrane potential (indicated by each trace) from the same cell patch clamped with an intracellular solution containing 4 mM Cl − and 0.5 mM QX-314 (see Methods). At a resting membrane potential of -42 mV, depolarizing potentials (▾) preceded hyperpolarizing components (▵). At -66 mV, only large depolarizing events were manifested while at -12 mV events consisted mainly of large hyperpolarizing potentials. B , EPSP cycle period versus IPSP cycle period during rhythmic bursting evoked by K + at a resting membrane potential of -42 mV. Data are from the cell shown in A. C , representative current clamp trace of spike activity recorded from a ventral interneurone (at -56 mV) following intracellular injection of a 500 ms depolarizing current pulse. In this case, QX-314 was omitted from the intracellular solution. Note that spike activity was maintained throughout the train without apparent accommodation.
    Figure Legend Snippet: Rhythmic activity induced by high-K + solution under current clamp recording A , rhythmic episodes evoked by 7 mM K + recorded at three different levels of resting membrane potential (indicated by each trace) from the same cell patch clamped with an intracellular solution containing 4 mM Cl − and 0.5 mM QX-314 (see Methods). At a resting membrane potential of -42 mV, depolarizing potentials (▾) preceded hyperpolarizing components (▵). At -66 mV, only large depolarizing events were manifested while at -12 mV events consisted mainly of large hyperpolarizing potentials. B , EPSP cycle period versus IPSP cycle period during rhythmic bursting evoked by K + at a resting membrane potential of -42 mV. Data are from the cell shown in A. C , representative current clamp trace of spike activity recorded from a ventral interneurone (at -56 mV) following intracellular injection of a 500 ms depolarizing current pulse. In this case, QX-314 was omitted from the intracellular solution. Note that spike activity was maintained throughout the train without apparent accommodation.

    Techniques Used: Activity Assay, Injection, Mass Spectrometry

    4) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    5) Product Images from "Presynaptic Actions of D2-Like Receptors in the Rat Cortico-Striato-Globus Pallidus Disynaptic Connection In Vitro"

    Article Title: Presynaptic Actions of D2-Like Receptors in the Rat Cortico-Striato-Globus Pallidus Disynaptic Connection In Vitro

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.90806.2008

    Triple stimulation with 3-ms interpulse interval of the cortex evoked inhibitory postsynaptic currents (IPSCs) in globus pallidus (GPe) neurons. The IPSCs in this and all subsequent figures were recorded with pipettes containing Cs 2 SO 4 (110 mM), tetraethylammonium (5 mM), and QX-314 (3 mM) to block potassium channels and from neurons voltage clamped at 0 mV. The stimulus artifacts in this and subsequent figures were truncated. A : examples of IPSCs evoked by 2 different stimulus intensities. Aa : stimulations with 63 μA evoked ∼7-pA IPSCs. Ab : increase in the stimulus intensity increased the amplitude of IPSCs and decreased the latency. B : plots of the amplitude (○) and the latency (•) of the IPSCs against the stimulus intensity. The data were obtained from the same neuron shown in A . Plots indicate that amplitudes and latencies of the IPSCs changed with the change in stimulus intensity. C : IPSCs from another GPe neuron. Ca : control in standard artificial cerebrospinal fluid (ACSF). Cb : bath application of 1,2,3,4-tetrahydro-6-nitro-2,3-dioxobenzo(f) quinoxaline-7sulfonamide (NBQX; 10 μM) and 3-(2-carboxypiperzin-4-yl)-propyl-1-phosponic acid (CPP; 30 μM) completely blocked the IPSCs. Cc : IPSCs partially recovered after a 20-min wash. Cd : subsequent application of gabazine (10 μM) also completely blocked the IPSCs.
    Figure Legend Snippet: Triple stimulation with 3-ms interpulse interval of the cortex evoked inhibitory postsynaptic currents (IPSCs) in globus pallidus (GPe) neurons. The IPSCs in this and all subsequent figures were recorded with pipettes containing Cs 2 SO 4 (110 mM), tetraethylammonium (5 mM), and QX-314 (3 mM) to block potassium channels and from neurons voltage clamped at 0 mV. The stimulus artifacts in this and subsequent figures were truncated. A : examples of IPSCs evoked by 2 different stimulus intensities. Aa : stimulations with 63 μA evoked ∼7-pA IPSCs. Ab : increase in the stimulus intensity increased the amplitude of IPSCs and decreased the latency. B : plots of the amplitude (○) and the latency (•) of the IPSCs against the stimulus intensity. The data were obtained from the same neuron shown in A . Plots indicate that amplitudes and latencies of the IPSCs changed with the change in stimulus intensity. C : IPSCs from another GPe neuron. Ca : control in standard artificial cerebrospinal fluid (ACSF). Cb : bath application of 1,2,3,4-tetrahydro-6-nitro-2,3-dioxobenzo(f) quinoxaline-7sulfonamide (NBQX; 10 μM) and 3-(2-carboxypiperzin-4-yl)-propyl-1-phosponic acid (CPP; 30 μM) completely blocked the IPSCs. Cc : IPSCs partially recovered after a 20-min wash. Cd : subsequent application of gabazine (10 μM) also completely blocked the IPSCs.

    Techniques Used: Mass Spectrometry, Blocking Assay, Conditioned Place Preference

    6) Product Images from "Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones"

    Article Title: Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.2000.t01-1-00231.x

    Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P
    Figure Legend Snippet: Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P

    Techniques Used: Mass Spectrometry

    7) Product Images from "Delimiting the Binding Site for Quaternary Ammonium Lidocaine Derivatives in the Acetylcholine Receptor Channel "

    Article Title: Delimiting the Binding Site for Quaternary Ammonium Lidocaine Derivatives in the Acetylcholine Receptor Channel

    Journal: The Journal of General Physiology

    doi:

    The effect of QX-314 on the irreversible inhibition of the ACh-induced current in two mutants. ( A and B ) αL258C. ( C and D ) αS248C. A and C show the test responses to ACh between applications of MTSEA in the presence of ACh and in the presence or absence of QX-314. B and D show the test currents as a function of cumulative exposure time to MTSEA. (▪) Initial responses and, in C and D , also extra final responses without any further application of MTSEA. (▵) MTSEA in the presence of ACh. (○) MTSEA in the presence of both ACh and QX-314. ACh was applied at 25 μM to αL258C and at 100 μM to αS248C. 1 mM MTSEA was applied in 3-s pulses to αL258C, and 1.5 mM MTSEA was applied in 15-s pulses to αS248C. QX-314 was applied at 1 mM to αL258C and at 0.2 mM to αS248C. The calibration bars are 3 s and 5 μA.
    Figure Legend Snippet: The effect of QX-314 on the irreversible inhibition of the ACh-induced current in two mutants. ( A and B ) αL258C. ( C and D ) αS248C. A and C show the test responses to ACh between applications of MTSEA in the presence of ACh and in the presence or absence of QX-314. B and D show the test currents as a function of cumulative exposure time to MTSEA. (▪) Initial responses and, in C and D , also extra final responses without any further application of MTSEA. (▵) MTSEA in the presence of ACh. (○) MTSEA in the presence of both ACh and QX-314. ACh was applied at 25 μM to αL258C and at 100 μM to αS248C. 1 mM MTSEA was applied in 3-s pulses to αL258C, and 1.5 mM MTSEA was applied in 15-s pulses to αS248C. QX-314 was applied at 1 mM to αL258C and at 0.2 mM to αS248C. The calibration bars are 3 s and 5 μA.

    Techniques Used: Inhibition

    The protection by QX-314 and QX-222 of Cys-substituted residues in M2 against reaction with MTSEA. For each mutant, the rate constant of the reaction in the presence of channel blocker and ACh ( k blocked ) and the rate constant of the reaction in the presence of ACh alone ( k open ) were determined as described in methods . The extent of protection is taken as 1 − ( k blocked / k open ). The means of two to four determinations and average errors or SEM are plotted. The lighter bars represent the protection by QX-314, and the darker bars represent the protection by QX-222. *ND.
    Figure Legend Snippet: The protection by QX-314 and QX-222 of Cys-substituted residues in M2 against reaction with MTSEA. For each mutant, the rate constant of the reaction in the presence of channel blocker and ACh ( k blocked ) and the rate constant of the reaction in the presence of ACh alone ( k open ) were determined as described in methods . The extent of protection is taken as 1 − ( k blocked / k open ). The means of two to four determinations and average errors or SEM are plotted. The lighter bars represent the protection by QX-314, and the darker bars represent the protection by QX-222. *ND.

    Techniques Used: Mutagenesis

    The dependence of protection on blocker concentration. The rate constant for the reaction of MTSEA in the presence of ACh at 10× EC 50 and of the indicated concentration of QX-314 is divided by the rate constant in the presence of ACh but absence of QX-314. The mutants are αV255C (•) and αS248C (▪). The arrows mark the interpolated values of the relative rate constants at the IC 50 s for QX-314 inhibition of the ACh-induced current (Table I ).
    Figure Legend Snippet: The dependence of protection on blocker concentration. The rate constant for the reaction of MTSEA in the presence of ACh at 10× EC 50 and of the indicated concentration of QX-314 is divided by the rate constant in the presence of ACh but absence of QX-314. The mutants are αV255C (•) and αS248C (▪). The arrows mark the interpolated values of the relative rate constants at the IC 50 s for QX-314 inhibition of the ACh-induced current (Table I ).

    Techniques Used: Concentration Assay, Relative Rate, Inhibition

    A model of QX-314 binding in the channel between two αM2 segments. The M2 segments are drawn as α helices. The dots represent the van der Waals surfaces. The drawing was made on a Silicon Graphics workstation running Insight II.
    Figure Legend Snippet: A model of QX-314 binding in the channel between two αM2 segments. The M2 segments are drawn as α helices. The dots represent the van der Waals surfaces. The drawing was made on a Silicon Graphics workstation running Insight II.

    Techniques Used: Binding Assay

    Structures of QX-314 and QX-222.
    Figure Legend Snippet: Structures of QX-314 and QX-222.

    Techniques Used:

    Inhibition of ACh-induced current in αL251C by QX-314 at different holding potentials. The top trace shows the holding potential. Both before and ∼3 s after the application of 25 μM ACh, the holding potential was stepped from −50 mV to −125, −75, −25, and back to −50 mV. The bottom trace shows the current. ( inset ) The peak ACh-induced current during the voltage steps minus the current during the voltage steps before ACh was added, with an expanded time axis. ( A ) No QX-314, ( B ) 0.1 mM QX-314, ( C ) 0.3 mM QX-314, ( D ) 1 mM QX-314. The four sections are part of a single experiment. Between the sections shown, the oocyte was washed and QX-314 at the indicated concentration was applied for 45 s before, and in addition to, the time indicated in the traces.
    Figure Legend Snippet: Inhibition of ACh-induced current in αL251C by QX-314 at different holding potentials. The top trace shows the holding potential. Both before and ∼3 s after the application of 25 μM ACh, the holding potential was stepped from −50 mV to −125, −75, −25, and back to −50 mV. The bottom trace shows the current. ( inset ) The peak ACh-induced current during the voltage steps minus the current during the voltage steps before ACh was added, with an expanded time axis. ( A ) No QX-314, ( B ) 0.1 mM QX-314, ( C ) 0.3 mM QX-314, ( D ) 1 mM QX-314. The four sections are part of a single experiment. Between the sections shown, the oocyte was washed and QX-314 at the indicated concentration was applied for 45 s before, and in addition to, the time indicated in the traces.

    Techniques Used: Inhibition, Concentration Assay

    The dependence on ACh of the protection by QX-314 against the irreversible inhibition by MTSEA. The mutant was αT244C. The log of the ACh-induced current is plotted against the cumulative time of application of MTSEA. ( A ) QX-314 added in the presence of ACh. ( B ) QX-314 added in the absence of ACh. (▪) Responses to 60 μM ACh before MTSEA was added. (○) Responses to ACh after 25-s applications of 2.5 μM MTSEA plus 60 μM ACh plus 0.1 mM QX-314. (▵) Responses to ACh after 25-s applications of 2.5 μM MTSEA plus 60 μM ACh. (▴) Responses to ACh after 25-s applications of 250 μM MTSEA. (•) Responses to ACh after 25-s applications of 250 μM MTSEA plus 1 mM QX-314.
    Figure Legend Snippet: The dependence on ACh of the protection by QX-314 against the irreversible inhibition by MTSEA. The mutant was αT244C. The log of the ACh-induced current is plotted against the cumulative time of application of MTSEA. ( A ) QX-314 added in the presence of ACh. ( B ) QX-314 added in the absence of ACh. (▪) Responses to 60 μM ACh before MTSEA was added. (○) Responses to ACh after 25-s applications of 2.5 μM MTSEA plus 60 μM ACh plus 0.1 mM QX-314. (▵) Responses to ACh after 25-s applications of 2.5 μM MTSEA plus 60 μM ACh. (▴) Responses to ACh after 25-s applications of 250 μM MTSEA. (•) Responses to ACh after 25-s applications of 250 μM MTSEA plus 1 mM QX-314.

    Techniques Used: Inhibition, Mutagenesis

    8) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    9) Product Images from "Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb"

    Article Title: Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.20-05-02011.2000

    Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.
    Figure Legend Snippet: Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.

    Techniques Used: Generated

    10) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    11) Product Images from "Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones"

    Article Title: Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.2000.t01-1-00231.x

    Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P
    Figure Legend Snippet: Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P

    Techniques Used: Mass Spectrometry

    12) Product Images from "Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb"

    Article Title: Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.20-05-02011.2000

    Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.
    Figure Legend Snippet: Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.

    Techniques Used: Generated

    13) Product Images from "Generation of rhythmic patterns of activity by ventral interneurones in rat organotypic spinal slice culture"

    Article Title: Generation of rhythmic patterns of activity by ventral interneurones in rat organotypic spinal slice culture

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.1999.0459t.x

    Rhythmic activity induced by high-K + solution under current clamp recording A , rhythmic episodes evoked by 7 mM K + recorded at three different levels of resting membrane potential (indicated by each trace) from the same cell patch clamped with an intracellular solution containing 4 mM Cl − and 0.5 mM QX-314 (see Methods). At a resting membrane potential of -42 mV, depolarizing potentials (▾) preceded hyperpolarizing components (▵). At -66 mV, only large depolarizing events were manifested while at -12 mV events consisted mainly of large hyperpolarizing potentials. B , EPSP cycle period versus IPSP cycle period during rhythmic bursting evoked by K + at a resting membrane potential of -42 mV. Data are from the cell shown in A. C , representative current clamp trace of spike activity recorded from a ventral interneurone (at -56 mV) following intracellular injection of a 500 ms depolarizing current pulse. In this case, QX-314 was omitted from the intracellular solution. Note that spike activity was maintained throughout the train without apparent accommodation.
    Figure Legend Snippet: Rhythmic activity induced by high-K + solution under current clamp recording A , rhythmic episodes evoked by 7 mM K + recorded at three different levels of resting membrane potential (indicated by each trace) from the same cell patch clamped with an intracellular solution containing 4 mM Cl − and 0.5 mM QX-314 (see Methods). At a resting membrane potential of -42 mV, depolarizing potentials (▾) preceded hyperpolarizing components (▵). At -66 mV, only large depolarizing events were manifested while at -12 mV events consisted mainly of large hyperpolarizing potentials. B , EPSP cycle period versus IPSP cycle period during rhythmic bursting evoked by K + at a resting membrane potential of -42 mV. Data are from the cell shown in A. C , representative current clamp trace of spike activity recorded from a ventral interneurone (at -56 mV) following intracellular injection of a 500 ms depolarizing current pulse. In this case, QX-314 was omitted from the intracellular solution. Note that spike activity was maintained throughout the train without apparent accommodation.

    Techniques Used: Activity Assay, Injection, Mass Spectrometry

    14) Product Images from "Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones"

    Article Title: Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.2000.t01-1-00231.x

    Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P
    Figure Legend Snippet: Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P

    Techniques Used: Mass Spectrometry

    15) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    16) Product Images from "Generation of rhythmic patterns of activity by ventral interneurones in rat organotypic spinal slice culture"

    Article Title: Generation of rhythmic patterns of activity by ventral interneurones in rat organotypic spinal slice culture

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.1999.0459t.x

    Rhythmic activity induced by high-K + solution under current clamp recording A , rhythmic episodes evoked by 7 mM K + recorded at three different levels of resting membrane potential (indicated by each trace) from the same cell patch clamped with an intracellular solution containing 4 mM Cl − and 0.5 mM QX-314 (see Methods). At a resting membrane potential of -42 mV, depolarizing potentials (▾) preceded hyperpolarizing components (▵). At -66 mV, only large depolarizing events were manifested while at -12 mV events consisted mainly of large hyperpolarizing potentials. B , EPSP cycle period versus IPSP cycle period during rhythmic bursting evoked by K + at a resting membrane potential of -42 mV. Data are from the cell shown in A. C , representative current clamp trace of spike activity recorded from a ventral interneurone (at -56 mV) following intracellular injection of a 500 ms depolarizing current pulse. In this case, QX-314 was omitted from the intracellular solution. Note that spike activity was maintained throughout the train without apparent accommodation.
    Figure Legend Snippet: Rhythmic activity induced by high-K + solution under current clamp recording A , rhythmic episodes evoked by 7 mM K + recorded at three different levels of resting membrane potential (indicated by each trace) from the same cell patch clamped with an intracellular solution containing 4 mM Cl − and 0.5 mM QX-314 (see Methods). At a resting membrane potential of -42 mV, depolarizing potentials (▾) preceded hyperpolarizing components (▵). At -66 mV, only large depolarizing events were manifested while at -12 mV events consisted mainly of large hyperpolarizing potentials. B , EPSP cycle period versus IPSP cycle period during rhythmic bursting evoked by K + at a resting membrane potential of -42 mV. Data are from the cell shown in A. C , representative current clamp trace of spike activity recorded from a ventral interneurone (at -56 mV) following intracellular injection of a 500 ms depolarizing current pulse. In this case, QX-314 was omitted from the intracellular solution. Note that spike activity was maintained throughout the train without apparent accommodation.

    Techniques Used: Activity Assay, Injection, Mass Spectrometry

    17) Product Images from "Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones"

    Article Title: Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.2000.t01-1-00231.x

    Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P
    Figure Legend Snippet: Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P

    Techniques Used: Mass Spectrometry

    18) Product Images from "Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb"

    Article Title: Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.20-05-02011.2000

    Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.
    Figure Legend Snippet: Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.

    Techniques Used: Generated

    19) Product Images from "The weaver Mutation Reverses the Function of Dopamine and GABA in Mouse Dopaminergic Neurons"

    Article Title: The weaver Mutation Reverses the Function of Dopamine and GABA in Mouse Dopaminergic Neurons

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.20-16-06013.2000

    The cation channel blockers QX-314 and ZD 7288 inhibit the DA- and baclofen-induced inward currents in wv/wv neurons. A , B , The DA-induced (100 μ m ) current was strongly inhibited by QX-314 (100 μ m ) and ZD 7288 (50 μ m ). C , D , QX-314
    Figure Legend Snippet: The cation channel blockers QX-314 and ZD 7288 inhibit the DA- and baclofen-induced inward currents in wv/wv neurons. A , B , The DA-induced (100 μ m ) current was strongly inhibited by QX-314 (100 μ m ) and ZD 7288 (50 μ m ). C , D , QX-314

    Techniques Used:

    20) Product Images from "Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb"

    Article Title: Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.20-05-02011.2000

    Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.
    Figure Legend Snippet: Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.

    Techniques Used: Generated

    21) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    22) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    23) Product Images from "Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology"

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    Journal: Frontiers in Neural Circuits

    doi: 10.3389/fncir.2012.00101

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.
    Figure Legend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Techniques Used: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Techniques Used: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.
    Figure Legend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Techniques Used: Injection, Activity Assay

    24) Product Images from "Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb"

    Article Title: Long-Lasting Depolarizations in Mitral Cells of the Rat Olfactory Bulb

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.20-05-02011.2000

    Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.
    Figure Legend Snippet: Somatic voltage-clamp recordings from a single mitral cell, depicting responses to orthodromic stimulation of the olfactory nerve ( A ), antidromic stimulation below the mitral cell layer ( B ), and spontaneous LLDs ( E ). Antidromic and orthodromic stimulation evoke highly reproducible and nearly identical LLDs (compare A , B ). This is consistent with LLDs being generated by interactions among M/T cells. In this cell, recorded without QX-314, antidromically evoked LLDs are preceded by an antidromic spike ( B ). To quantify the variance of the evoked LLDs, we plotted the SDs ( SDev ) of the averaged traces ( C , D ). The largest variance occurs during the late component of the rising phase of the LLDs (delineated by the vertical dashed lines ). This is also consistent with the hypothesis that LLDs are generated by multiple interactions among M/T cells. The plot in C is shown at higher magnification in the inset . Filled circles correspond to the time points used in the text to represent the group variance data (at 20 msec after stimulus, the peak of the SD, and the peak of the LLD). F , Averaged spontaneous LLDs and their variance obtained from the traces depicted in E . Individual traces were aligned relative to the onset of their fastest component ( F, first vertical dashed line ); as in the evoked LLDs, variability is largest before the peak. G , Autocorrelogram of spontaneous LLDs ( thick line ) and peristimulus histogram ( bars ) of evoked LLDs (excluding the initial LLD) shows the similar refractory period for both spontaneous and evoked responses.

    Techniques Used: Generated

    25) Product Images from "Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones"

    Article Title: Novel action of BAPTA series chelators on intrinsic K+ currents in rat hippocampal neurones

    Journal: The Journal of Physiology

    doi: 10.1111/j.1469-7793.2000.t01-1-00231.x

    Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P
    Figure Legend Snippet: Action of BAPTA series chelators on holding current The histogram plots maximal outward current recorded in the presence of BAPTA series chelators under various conditions compared with control data without buffer (con, n = 10) and the I sAHP evoked by an 80 ms depolarization from −60 to −10 mV ( n = 21). The dibromo BAPTA (1 mM) data set includes 50 % Ca 2+ loaded (Ca 2+ , n = 5), with 10 mM EGTA (EGTA, n = 5), with 100 μM 8-Br cAMP (cAMP, n = 7), without depolarization (-depol) plus and minus 10 mM QX-314 ( n = 9 and 5, respectively). Data for diazo-2 (2 mM, n = 20) and BAPTA itself (2 mM, n = 6) are also shown. The cAMP and QX-314 data are not significantly different from control data. Values of total conductance ( G ) were calculated after correcting for voltage offsets and are presented under the bars for the chelator data which are significantly different from control ( P

    Techniques Used: Mass Spectrometry

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    Alomone Labs qx314
    Effects of <t>QX314</t> on Dopamine modulation of Glutamatergic EPSCs. A : Comparison of the average change in amplitude of the early component of the glutamatergic EPSCs (in pA) measured at −60 mV, on superfusion of DA in the presence and absence of QX314. The solid bar represents the average response in 80 neurons (n = 80), in the presence of QX314 and the dashed bar represents the average of 8 neurons (n = 8) in the absence of QX314. No statistically significant difference was found between the two groups (Paired Student’s t test p
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    Effects of QX314 on Dopamine modulation of Glutamatergic EPSCs. A : Comparison of the average change in amplitude of the early component of the glutamatergic EPSCs (in pA) measured at −60 mV, on superfusion of DA in the presence and absence of QX314. The solid bar represents the average response in 80 neurons (n = 80), in the presence of QX314 and the dashed bar represents the average of 8 neurons (n = 8) in the absence of QX314. No statistically significant difference was found between the two groups (Paired Student’s t test p

    Journal: PLoS ONE

    Article Title: Dopamine Preferentially Inhibits NMDA Receptor-Mediated EPSCs by Acting on Presynaptic D1 Receptors in Nucleus Accumbens during Postnatal Development

    doi: 10.1371/journal.pone.0086970

    Figure Lengend Snippet: Effects of QX314 on Dopamine modulation of Glutamatergic EPSCs. A : Comparison of the average change in amplitude of the early component of the glutamatergic EPSCs (in pA) measured at −60 mV, on superfusion of DA in the presence and absence of QX314. The solid bar represents the average response in 80 neurons (n = 80), in the presence of QX314 and the dashed bar represents the average of 8 neurons (n = 8) in the absence of QX314. No statistically significant difference was found between the two groups (Paired Student’s t test p

    Article Snippet: QX314 (5 mM; Alomone Laboratories, Jerusalem, Israel) was routinely added to the recording pipette solution to prevent voltage-sensitive Na+ channels from generating action potentials.

    Techniques:

    Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Journal: Frontiers in Neural Circuits

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    doi: 10.3389/fncir.2012.00101

    Figure Lengend Snippet: Setup and experimental strategies used to prevent spikes during functional mapping of synaptic inputs in vivo . (A) Schematic of the preparation showing simultaneous intracellular recording and dendritic calcium imaging in a layer 2/3 pyramidal neuron in vivo . (B) The electrode enters the cortex through a hole in the coverglass. (C) Spikes are suppressed either by injection of a constant negative current to hyperpolarize the neuron below spike threshold (left), or by including QX-314 in the pipette solution (middle), or by iontophoresis of GABA from an adjacent patch pipette glued to the sharp microelectrode (right). (D) Fabrication of the double barrel electrode used for simultaneous intracellular recording and extracellular iontophoresis of GABA: a bent patch pipette and a straight sharp microelectrode are bonded together by UV-sensitive glue. Bottom right, micrograph of the two tips of a completed double barrel electrode (inter-tip distance: 15 μm). Also see Figure A4 in Appendix for detailed steps on the assembly of double barrel electrodes.

    Article Snippet: In a given neuron, visually evoked spikes were suppressed by one of the following strategies: (1) hyperpolarization by injection of a constant negative current (n = 5 neurons); (2) blockade of voltage-sensitive sodium channels by including QX-314 in the electrode solution (n = 7 neurons); or (3) GABA iontophoresis near the soma of the intracellularly recorded neuron from an iontophoresis electrode glued to the sharp electrode (n = 4 neurons).

    Techniques: Functional Assay, In Vivo, Imaging, Injection, Transferring

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Journal: Frontiers in Neural Circuits

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    doi: 10.3389/fncir.2012.00101

    Figure Lengend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314. (A) Spikes were suppressed by QX-314 (100 mM) contained in the intra-electrode solution. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 265 μm below the pia, and visual responses were imaged in a basal dendrite (red box, depth: 275 μm below the pia). (C) Membrane potential responses to eight stimulus directions. (D) Corresponding calcium responses imaged in the dendrite highlighted in (B) . Thick red trace: average of three repeats. (E) Expanded time course of the membrane potential response to the preferred stimulus direction. Each cycle of the drifting grating visual stimulus elicited QX-insensitive spikelets (arrowheads). (F) Expanded time course showing the shape of the QX-insensitive spikelets, which are smaller and slower than sodium spikes. No current was injected during this recording.

    Article Snippet: In a given neuron, visually evoked spikes were suppressed by one of the following strategies: (1) hyperpolarization by injection of a constant negative current (n = 5 neurons); (2) blockade of voltage-sensitive sodium channels by including QX-314 in the electrode solution (n = 7 neurons); or (3) GABA iontophoresis near the soma of the intracellularly recorded neuron from an iontophoresis electrode glued to the sharp electrode (n = 4 neurons).

    Techniques: Injection

    Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Journal: Frontiers in Neural Circuits

    Article Title: Strategies for mapping synaptic inputs on dendrites in vivo by combining two-photon microscopy, sharp intracellular recording, and pharmacology

    doi: 10.3389/fncir.2012.00101

    Figure Lengend Snippet: Dendritic visual responses during pharmacological blockade of spikes in a neuron loaded with QX-314 and simultaneous injection of hyperpolarizing current. (A) 50 mM QX-314 was included in the sharp micropipette, and −0.7 nA current was injected. (B) Z-projection of the reconstructed somato-dendritic tree. The soma was located 280 μm below the pia. Visual responses were imaged in an apical dendrite (red box, depth: 185 μm below the pia). (C) Membrane potential responses to visual stimuli moving in eight different directions (five repeats). I DC = −0.7 nA. Note the absence of spikes. (D) Corresponding visual responses in the apical dendrite highlighted in (B) . Thick red trace: average of five repeats. (E) Spontaneous membrane potential activity for different amplitudes of constant negative current. QX-insensitive spikelets are progressively suppressed as the neuron is further hyperpolarized with −0.4 to −0.7 nA current injection. (F) Expanded time course of the membrane potential response at the preferred direction during negative current injection (−0.7 nA). Note the absence of QX-insensitive spikelets.

    Article Snippet: In a given neuron, visually evoked spikes were suppressed by one of the following strategies: (1) hyperpolarization by injection of a constant negative current (n = 5 neurons); (2) blockade of voltage-sensitive sodium channels by including QX-314 in the electrode solution (n = 7 neurons); or (3) GABA iontophoresis near the soma of the intracellularly recorded neuron from an iontophoresis electrode glued to the sharp electrode (n = 4 neurons).

    Techniques: Injection, Activity Assay