nbqx  (Alomone Labs)


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    Alomone Labs nbqx
    Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM <t>NBQX.</t> Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o <t>+</t> <t>mannitol,</t> mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.
    Nbqx, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium"

    Article Title: Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2022.930384

    Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM NBQX. Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o + mannitol, mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.
    Figure Legend Snippet: Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM NBQX. Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o + mannitol, mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.

    Techniques Used: Blocking Assay, Patch Clamp

    Addition of DL-AP5 attenuates volume-related effects on neuronal excitability. (A) Generally, the amount of holding current to maintain voltage-clamp at –70 mV was reduced in ∧ [K + ] o + mannitol compared to ∧ [K + ] o alone in all experiments. Holding currents for mixed AMPA + NMDA receptor experiments (blue, n = 9) and those isolating NMDA receptor currents (+NBQX) (green, n = 13) appeared remarkably similar and had no points during which they were significantly different. Holding currents recorded during the NMDA receptor inhibition experiments (+NBQX/+DL-AP5) were less negative overall and were significantly less negative following the second ∧ [K + ] o + mannitol co-application period (yellow, n = 8). (B) Comparison of resting membrane potentials for the NMDA receptor isolation and NMDA receptor inhibition experiments. Shifts in resting membrane potential indicated that cells became depolarized relative to baseline during application of ∧ [K + ] o , while co-application of ∧ [K + ] o + mannitol triggered slight hyperpolarizing shifts. Resting membrane potentials recorded during the NMDAR inhibition experiments (yellow, n = 8) indicated significantly less depolarization compared to the NMDAR isolation experiments following the initial application of ∧ [K + ] o alone, the mannitol wash period, and the second co-application of ∧ [K + ] o + mannitol (green, n = 13). * p < 0.05 and *** p < 0.001.
    Figure Legend Snippet: Addition of DL-AP5 attenuates volume-related effects on neuronal excitability. (A) Generally, the amount of holding current to maintain voltage-clamp at –70 mV was reduced in ∧ [K + ] o + mannitol compared to ∧ [K + ] o alone in all experiments. Holding currents for mixed AMPA + NMDA receptor experiments (blue, n = 9) and those isolating NMDA receptor currents (+NBQX) (green, n = 13) appeared remarkably similar and had no points during which they were significantly different. Holding currents recorded during the NMDA receptor inhibition experiments (+NBQX/+DL-AP5) were less negative overall and were significantly less negative following the second ∧ [K + ] o + mannitol co-application period (yellow, n = 8). (B) Comparison of resting membrane potentials for the NMDA receptor isolation and NMDA receptor inhibition experiments. Shifts in resting membrane potential indicated that cells became depolarized relative to baseline during application of ∧ [K + ] o , while co-application of ∧ [K + ] o + mannitol triggered slight hyperpolarizing shifts. Resting membrane potentials recorded during the NMDAR inhibition experiments (yellow, n = 8) indicated significantly less depolarization compared to the NMDAR isolation experiments following the initial application of ∧ [K + ] o alone, the mannitol wash period, and the second co-application of ∧ [K + ] o + mannitol (green, n = 13). * p < 0.05 and *** p < 0.001.

    Techniques Used: Inhibition, Isolation

    DL-AP5 substantially attenuates NMDA receptor currents during application of ∧ [K + ] o . Approximately 1-min section of recording taken from the mannitol wash period for an experiment conducted in NBQX without DL-AP5 (A) compared to the mannitol wash period with 50 μM DL-AP5 (B) . Note the number of large SIC-like events in the absence, but not the presence, of DL-AP5. (C) Frequency of all NMDA receptor events was significantly lower in DL-AP5 (yellow, n = 8) compared to + NBQX alone (green, n = 13) during ∧ [K + ] o application both with and without mannitol. (D) When separating events into individual 10-min recording periods, NMDA receptor events were significantly reduced during the mannitol wash period and second ∧ [K + ] o + mannitol application period. (E) DL-AP5 also significantly inhibited the occurrence of SICs either with or without mannitol. (F) When grouping frequencies of SICs into 10-min time bins, DL-AP5 significantly inhibited SICs during the mannitol wash period and second ∧ [K + ] o + mannitol co-application period. SICs were blocked completely during the second ∧ [K + ] o + mannitol co-application period. * p < 0.05, ** p < 0.01, and *** p < 0.001.
    Figure Legend Snippet: DL-AP5 substantially attenuates NMDA receptor currents during application of ∧ [K + ] o . Approximately 1-min section of recording taken from the mannitol wash period for an experiment conducted in NBQX without DL-AP5 (A) compared to the mannitol wash period with 50 μM DL-AP5 (B) . Note the number of large SIC-like events in the absence, but not the presence, of DL-AP5. (C) Frequency of all NMDA receptor events was significantly lower in DL-AP5 (yellow, n = 8) compared to + NBQX alone (green, n = 13) during ∧ [K + ] o application both with and without mannitol. (D) When separating events into individual 10-min recording periods, NMDA receptor events were significantly reduced during the mannitol wash period and second ∧ [K + ] o + mannitol application period. (E) DL-AP5 also significantly inhibited the occurrence of SICs either with or without mannitol. (F) When grouping frequencies of SICs into 10-min time bins, DL-AP5 significantly inhibited SICs during the mannitol wash period and second ∧ [K + ] o + mannitol co-application period. SICs were blocked completely during the second ∧ [K + ] o + mannitol co-application period. * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Techniques Used:

    nbqx  (Alomone Labs)


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    Alomone Labs nbqx
    Nbqx, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM <t>NBQX.</t> Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o <t>+</t> <t>mannitol,</t> mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.
    Nbqx, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium"

    Article Title: Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium

    Journal: Frontiers in Cellular Neuroscience

    doi: 10.3389/fncel.2022.930384

    Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM NBQX. Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o + mannitol, mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.
    Figure Legend Snippet: Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM NBQX. Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o + mannitol, mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.

    Techniques Used: Blocking Assay, Patch Clamp

    Addition of DL-AP5 attenuates volume-related effects on neuronal excitability. (A) Generally, the amount of holding current to maintain voltage-clamp at –70 mV was reduced in ∧ [K + ] o + mannitol compared to ∧ [K + ] o alone in all experiments. Holding currents for mixed AMPA + NMDA receptor experiments (blue, n = 9) and those isolating NMDA receptor currents (+NBQX) (green, n = 13) appeared remarkably similar and had no points during which they were significantly different. Holding currents recorded during the NMDA receptor inhibition experiments (+NBQX/+DL-AP5) were less negative overall and were significantly less negative following the second ∧ [K + ] o + mannitol co-application period (yellow, n = 8). (B) Comparison of resting membrane potentials for the NMDA receptor isolation and NMDA receptor inhibition experiments. Shifts in resting membrane potential indicated that cells became depolarized relative to baseline during application of ∧ [K + ] o , while co-application of ∧ [K + ] o + mannitol triggered slight hyperpolarizing shifts. Resting membrane potentials recorded during the NMDAR inhibition experiments (yellow, n = 8) indicated significantly less depolarization compared to the NMDAR isolation experiments following the initial application of ∧ [K + ] o alone, the mannitol wash period, and the second co-application of ∧ [K + ] o + mannitol (green, n = 13). * p < 0.05 and *** p < 0.001.
    Figure Legend Snippet: Addition of DL-AP5 attenuates volume-related effects on neuronal excitability. (A) Generally, the amount of holding current to maintain voltage-clamp at –70 mV was reduced in ∧ [K + ] o + mannitol compared to ∧ [K + ] o alone in all experiments. Holding currents for mixed AMPA + NMDA receptor experiments (blue, n = 9) and those isolating NMDA receptor currents (+NBQX) (green, n = 13) appeared remarkably similar and had no points during which they were significantly different. Holding currents recorded during the NMDA receptor inhibition experiments (+NBQX/+DL-AP5) were less negative overall and were significantly less negative following the second ∧ [K + ] o + mannitol co-application period (yellow, n = 8). (B) Comparison of resting membrane potentials for the NMDA receptor isolation and NMDA receptor inhibition experiments. Shifts in resting membrane potential indicated that cells became depolarized relative to baseline during application of ∧ [K + ] o , while co-application of ∧ [K + ] o + mannitol triggered slight hyperpolarizing shifts. Resting membrane potentials recorded during the NMDAR inhibition experiments (yellow, n = 8) indicated significantly less depolarization compared to the NMDAR isolation experiments following the initial application of ∧ [K + ] o alone, the mannitol wash period, and the second co-application of ∧ [K + ] o + mannitol (green, n = 13). * p < 0.05 and *** p < 0.001.

    Techniques Used: Inhibition, Isolation

    DL-AP5 substantially attenuates NMDA receptor currents during application of ∧ [K + ] o . Approximately 1-min section of recording taken from the mannitol wash period for an experiment conducted in NBQX without DL-AP5 (A) compared to the mannitol wash period with 50 μM DL-AP5 (B) . Note the number of large SIC-like events in the absence, but not the presence, of DL-AP5. (C) Frequency of all NMDA receptor events was significantly lower in DL-AP5 (yellow, n = 8) compared to + NBQX alone (green, n = 13) during ∧ [K + ] o application both with and without mannitol. (D) When separating events into individual 10-min recording periods, NMDA receptor events were significantly reduced during the mannitol wash period and second ∧ [K + ] o + mannitol application period. (E) DL-AP5 also significantly inhibited the occurrence of SICs either with or without mannitol. (F) When grouping frequencies of SICs into 10-min time bins, DL-AP5 significantly inhibited SICs during the mannitol wash period and second ∧ [K + ] o + mannitol co-application period. SICs were blocked completely during the second ∧ [K + ] o + mannitol co-application period. * p < 0.05, ** p < 0.01, and *** p < 0.001.
    Figure Legend Snippet: DL-AP5 substantially attenuates NMDA receptor currents during application of ∧ [K + ] o . Approximately 1-min section of recording taken from the mannitol wash period for an experiment conducted in NBQX without DL-AP5 (A) compared to the mannitol wash period with 50 μM DL-AP5 (B) . Note the number of large SIC-like events in the absence, but not the presence, of DL-AP5. (C) Frequency of all NMDA receptor events was significantly lower in DL-AP5 (yellow, n = 8) compared to + NBQX alone (green, n = 13) during ∧ [K + ] o application both with and without mannitol. (D) When separating events into individual 10-min recording periods, NMDA receptor events were significantly reduced during the mannitol wash period and second ∧ [K + ] o + mannitol application period. (E) DL-AP5 also significantly inhibited the occurrence of SICs either with or without mannitol. (F) When grouping frequencies of SICs into 10-min time bins, DL-AP5 significantly inhibited SICs during the mannitol wash period and second ∧ [K + ] o + mannitol co-application period. SICs were blocked completely during the second ∧ [K + ] o + mannitol co-application period. * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Techniques Used:

    nbqx  (Alomone Labs)


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    Alomone Labs nbqx
    Nbqx, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs nbqx
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    Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM <t>NBQX.</t> Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o <t>+</t> <t>mannitol,</t> mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.
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    Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM NBQX. Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o + mannitol, mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium

    doi: 10.3389/fncel.2022.930384

    Figure Lengend Snippet: Application of hyperosmolar ∧ [K + ] o triggers changes in neuron membrane excitability despite block of AMPARs. (A) Representative recording of a neuron voltage-clamped at –70 mV in Mg 2+ -free ACSF + 1 μM TTX and 10 μM NBQX. Gaps in recordings coincide with periodic measurement membrane potential and holding current in order to assess patch-clamp quality between alternating 10-min ACSF treatments: control ACSF, ∧ [K + ] o , ∧ [K + ] o + mannitol, mannitol wash, and ∧ [K + ] o + mannitol. (B) Section of recording showing 10-min mannitol wash and 10-min second application of ∧ [K + ] o + mannitol. Inset shows the value of the holding current measured at the end of each application period ( n = 13). (C) An ∼ 4-min section of the recording in (B) , showing events during the mannitol wash period in ∧ [K + ] o . (D) An ∼ 4-min section of the recording in (B) , showing events during the second ∧ [K + ] o + mannitol application. Note the large SICs during the mannitol wash period compared to the smaller mEPSCs present during the second ∧ [K + ] o + mannitol co-application. (E) A comparison of averaged SIC traces from the first application of ∧ [K + ] o with the first ∧ [K + ] o + mannitol co-application (left) and between the mannitol wash and the second ∧ [K + ] o + mannitol co-application (right). * p < 0.05.

    Article Snippet: Additional experiments in ∧ [K + ] o ACSF + mannitol as described above were performed in the presence of 10 μM NBQX (Alomone Labs) to block AMPA receptor currents (NMDA receptor isolation), or with the addition of 50 μL DL-AP5 (Abcam) to block remaining NMDA receptor currents (NMDA receptor inhibition).

    Techniques: Blocking Assay, Patch Clamp

    Addition of DL-AP5 attenuates volume-related effects on neuronal excitability. (A) Generally, the amount of holding current to maintain voltage-clamp at –70 mV was reduced in ∧ [K + ] o + mannitol compared to ∧ [K + ] o alone in all experiments. Holding currents for mixed AMPA + NMDA receptor experiments (blue, n = 9) and those isolating NMDA receptor currents (+NBQX) (green, n = 13) appeared remarkably similar and had no points during which they were significantly different. Holding currents recorded during the NMDA receptor inhibition experiments (+NBQX/+DL-AP5) were less negative overall and were significantly less negative following the second ∧ [K + ] o + mannitol co-application period (yellow, n = 8). (B) Comparison of resting membrane potentials for the NMDA receptor isolation and NMDA receptor inhibition experiments. Shifts in resting membrane potential indicated that cells became depolarized relative to baseline during application of ∧ [K + ] o , while co-application of ∧ [K + ] o + mannitol triggered slight hyperpolarizing shifts. Resting membrane potentials recorded during the NMDAR inhibition experiments (yellow, n = 8) indicated significantly less depolarization compared to the NMDAR isolation experiments following the initial application of ∧ [K + ] o alone, the mannitol wash period, and the second co-application of ∧ [K + ] o + mannitol (green, n = 13). * p < 0.05 and *** p < 0.001.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium

    doi: 10.3389/fncel.2022.930384

    Figure Lengend Snippet: Addition of DL-AP5 attenuates volume-related effects on neuronal excitability. (A) Generally, the amount of holding current to maintain voltage-clamp at –70 mV was reduced in ∧ [K + ] o + mannitol compared to ∧ [K + ] o alone in all experiments. Holding currents for mixed AMPA + NMDA receptor experiments (blue, n = 9) and those isolating NMDA receptor currents (+NBQX) (green, n = 13) appeared remarkably similar and had no points during which they were significantly different. Holding currents recorded during the NMDA receptor inhibition experiments (+NBQX/+DL-AP5) were less negative overall and were significantly less negative following the second ∧ [K + ] o + mannitol co-application period (yellow, n = 8). (B) Comparison of resting membrane potentials for the NMDA receptor isolation and NMDA receptor inhibition experiments. Shifts in resting membrane potential indicated that cells became depolarized relative to baseline during application of ∧ [K + ] o , while co-application of ∧ [K + ] o + mannitol triggered slight hyperpolarizing shifts. Resting membrane potentials recorded during the NMDAR inhibition experiments (yellow, n = 8) indicated significantly less depolarization compared to the NMDAR isolation experiments following the initial application of ∧ [K + ] o alone, the mannitol wash period, and the second co-application of ∧ [K + ] o + mannitol (green, n = 13). * p < 0.05 and *** p < 0.001.

    Article Snippet: Additional experiments in ∧ [K + ] o ACSF + mannitol as described above were performed in the presence of 10 μM NBQX (Alomone Labs) to block AMPA receptor currents (NMDA receptor isolation), or with the addition of 50 μL DL-AP5 (Abcam) to block remaining NMDA receptor currents (NMDA receptor inhibition).

    Techniques: Inhibition, Isolation

    DL-AP5 substantially attenuates NMDA receptor currents during application of ∧ [K + ] o . Approximately 1-min section of recording taken from the mannitol wash period for an experiment conducted in NBQX without DL-AP5 (A) compared to the mannitol wash period with 50 μM DL-AP5 (B) . Note the number of large SIC-like events in the absence, but not the presence, of DL-AP5. (C) Frequency of all NMDA receptor events was significantly lower in DL-AP5 (yellow, n = 8) compared to + NBQX alone (green, n = 13) during ∧ [K + ] o application both with and without mannitol. (D) When separating events into individual 10-min recording periods, NMDA receptor events were significantly reduced during the mannitol wash period and second ∧ [K + ] o + mannitol application period. (E) DL-AP5 also significantly inhibited the occurrence of SICs either with or without mannitol. (F) When grouping frequencies of SICs into 10-min time bins, DL-AP5 significantly inhibited SICs during the mannitol wash period and second ∧ [K + ] o + mannitol co-application period. SICs were blocked completely during the second ∧ [K + ] o + mannitol co-application period. * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Contributions of Astrocyte and Neuronal Volume to CA1 Neuron Excitability Changes in Elevated Extracellular Potassium

    doi: 10.3389/fncel.2022.930384

    Figure Lengend Snippet: DL-AP5 substantially attenuates NMDA receptor currents during application of ∧ [K + ] o . Approximately 1-min section of recording taken from the mannitol wash period for an experiment conducted in NBQX without DL-AP5 (A) compared to the mannitol wash period with 50 μM DL-AP5 (B) . Note the number of large SIC-like events in the absence, but not the presence, of DL-AP5. (C) Frequency of all NMDA receptor events was significantly lower in DL-AP5 (yellow, n = 8) compared to + NBQX alone (green, n = 13) during ∧ [K + ] o application both with and without mannitol. (D) When separating events into individual 10-min recording periods, NMDA receptor events were significantly reduced during the mannitol wash period and second ∧ [K + ] o + mannitol application period. (E) DL-AP5 also significantly inhibited the occurrence of SICs either with or without mannitol. (F) When grouping frequencies of SICs into 10-min time bins, DL-AP5 significantly inhibited SICs during the mannitol wash period and second ∧ [K + ] o + mannitol co-application period. SICs were blocked completely during the second ∧ [K + ] o + mannitol co-application period. * p < 0.05, ** p < 0.01, and *** p < 0.001.

    Article Snippet: Additional experiments in ∧ [K + ] o ACSF + mannitol as described above were performed in the presence of 10 μM NBQX (Alomone Labs) to block AMPA receptor currents (NMDA receptor isolation), or with the addition of 50 μL DL-AP5 (Abcam) to block remaining NMDA receptor currents (NMDA receptor inhibition).

    Techniques: