α pompilidotoxin  (Alomone Labs)


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    Structured Review

    Alomone Labs α pompilidotoxin
    Neuronal stimulation triggers glycolysis in response to energy demand from ion pumping. a. Left: Representative trace of Peredox and RCaMP1h lifetimes simultaneously recorded in a DGC from an acute hippocampal slice. The slice was superfused with 2 µM GSK-2837808A for at least 30 min before the experiment, and the LDH inhibitor was kept in the ACSF during the experiment. The ACSF also contained 1 mM EGTA to reinforce Ca 2+ removal in the nominal 0Ca 2+ condition (but [Ca 2+ ] in the control ACSF was accordingly adjusted to a free concentration of 2 mM, as in any other experiment). Effective Ca 2+ removal was confirmed by the absence of a RCaMP1h spike upon stimulation. The Peredox lifetime at baseline, and the metabolic transients in response to neuronal stimulation, were recorded after substituting the bath solution with a 0Ca 2+ ACSF (to obtain Na + -only NADH CYT responses), and the further application of 10 µM <t>α-pompilidotoxin</t> (α-Pmtx, a toxin that prevents voltage-gated Na + channel inactivation). Right: The Na + -only NADH CYT transient was increased in the presence of α-pompilidotoxin. The Peredox lifetime change in response to stimulation was diminished in the absence of Ca 2+ but bounced back to higher amplitudes by increasing Na + influx. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test (N neurons = 35, N slices = 5 and N mice = 3). For all panels, only neurons showing an initial ΔPeredox lifetime response ≥0.05 ns were included for analysis. b. Representative trace of Peredox and RCaMP1h lifetimes in a DGC stimulated with trains of 100 and 200 electrical pulses. The two-stimulation protocol was also performed in 0Ca 2+ ACSF before and after the application of 3 µM α-pompilidotoxin. The latter condition was followed by the application of the Na + /K + ATPase inhibitor strophanthidin. As in (a) , the slices were exposed to the LDH inhibitor GSK-2837808A from 30 min prior, until the end of the experiment. Likewise, all the solutions contained 1 mM EGTA. c. Comparison of the Peredox lifetime changes in response to both stimulation paradigms (100 or 200 pulses) among the conditions in (b) . The NADH CYT transients in control condition was different from the other conditions (the discontinuous line for the associated p-value applies to all comparisons). The Na + -only NADH CYT responses recorded in 0Ca 2+ ACSF were increased slightly but significantly increased by the application of 3 µM α-pompilidotoxin, an effect that was reversed by strophanthidin. The data were compared using a non-parametric repeated measures ANOVA (Friedman test) with a Dunn post-test (N neurons = 66, N slices = 10 and N mice = 5).
    α Pompilidotoxin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/α pompilidotoxin/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    α pompilidotoxin - by Bioz Stars, 2022-07
    93/100 stars

    Images

    1) Product Images from "The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle"

    Article Title: The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle

    Journal: bioRxiv

    doi: 10.1101/2020.11.16.385526

    Neuronal stimulation triggers glycolysis in response to energy demand from ion pumping. a. Left: Representative trace of Peredox and RCaMP1h lifetimes simultaneously recorded in a DGC from an acute hippocampal slice. The slice was superfused with 2 µM GSK-2837808A for at least 30 min before the experiment, and the LDH inhibitor was kept in the ACSF during the experiment. The ACSF also contained 1 mM EGTA to reinforce Ca 2+ removal in the nominal 0Ca 2+ condition (but [Ca 2+ ] in the control ACSF was accordingly adjusted to a free concentration of 2 mM, as in any other experiment). Effective Ca 2+ removal was confirmed by the absence of a RCaMP1h spike upon stimulation. The Peredox lifetime at baseline, and the metabolic transients in response to neuronal stimulation, were recorded after substituting the bath solution with a 0Ca 2+ ACSF (to obtain Na + -only NADH CYT responses), and the further application of 10 µM α-pompilidotoxin (α-Pmtx, a toxin that prevents voltage-gated Na + channel inactivation). Right: The Na + -only NADH CYT transient was increased in the presence of α-pompilidotoxin. The Peredox lifetime change in response to stimulation was diminished in the absence of Ca 2+ but bounced back to higher amplitudes by increasing Na + influx. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test (N neurons = 35, N slices = 5 and N mice = 3). For all panels, only neurons showing an initial ΔPeredox lifetime response ≥0.05 ns were included for analysis. b. Representative trace of Peredox and RCaMP1h lifetimes in a DGC stimulated with trains of 100 and 200 electrical pulses. The two-stimulation protocol was also performed in 0Ca 2+ ACSF before and after the application of 3 µM α-pompilidotoxin. The latter condition was followed by the application of the Na + /K + ATPase inhibitor strophanthidin. As in (a) , the slices were exposed to the LDH inhibitor GSK-2837808A from 30 min prior, until the end of the experiment. Likewise, all the solutions contained 1 mM EGTA. c. Comparison of the Peredox lifetime changes in response to both stimulation paradigms (100 or 200 pulses) among the conditions in (b) . The NADH CYT transients in control condition was different from the other conditions (the discontinuous line for the associated p-value applies to all comparisons). The Na + -only NADH CYT responses recorded in 0Ca 2+ ACSF were increased slightly but significantly increased by the application of 3 µM α-pompilidotoxin, an effect that was reversed by strophanthidin. The data were compared using a non-parametric repeated measures ANOVA (Friedman test) with a Dunn post-test (N neurons = 66, N slices = 10 and N mice = 5).
    Figure Legend Snippet: Neuronal stimulation triggers glycolysis in response to energy demand from ion pumping. a. Left: Representative trace of Peredox and RCaMP1h lifetimes simultaneously recorded in a DGC from an acute hippocampal slice. The slice was superfused with 2 µM GSK-2837808A for at least 30 min before the experiment, and the LDH inhibitor was kept in the ACSF during the experiment. The ACSF also contained 1 mM EGTA to reinforce Ca 2+ removal in the nominal 0Ca 2+ condition (but [Ca 2+ ] in the control ACSF was accordingly adjusted to a free concentration of 2 mM, as in any other experiment). Effective Ca 2+ removal was confirmed by the absence of a RCaMP1h spike upon stimulation. The Peredox lifetime at baseline, and the metabolic transients in response to neuronal stimulation, were recorded after substituting the bath solution with a 0Ca 2+ ACSF (to obtain Na + -only NADH CYT responses), and the further application of 10 µM α-pompilidotoxin (α-Pmtx, a toxin that prevents voltage-gated Na + channel inactivation). Right: The Na + -only NADH CYT transient was increased in the presence of α-pompilidotoxin. The Peredox lifetime change in response to stimulation was diminished in the absence of Ca 2+ but bounced back to higher amplitudes by increasing Na + influx. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test (N neurons = 35, N slices = 5 and N mice = 3). For all panels, only neurons showing an initial ΔPeredox lifetime response ≥0.05 ns were included for analysis. b. Representative trace of Peredox and RCaMP1h lifetimes in a DGC stimulated with trains of 100 and 200 electrical pulses. The two-stimulation protocol was also performed in 0Ca 2+ ACSF before and after the application of 3 µM α-pompilidotoxin. The latter condition was followed by the application of the Na + /K + ATPase inhibitor strophanthidin. As in (a) , the slices were exposed to the LDH inhibitor GSK-2837808A from 30 min prior, until the end of the experiment. Likewise, all the solutions contained 1 mM EGTA. c. Comparison of the Peredox lifetime changes in response to both stimulation paradigms (100 or 200 pulses) among the conditions in (b) . The NADH CYT transients in control condition was different from the other conditions (the discontinuous line for the associated p-value applies to all comparisons). The Na + -only NADH CYT responses recorded in 0Ca 2+ ACSF were increased slightly but significantly increased by the application of 3 µM α-pompilidotoxin, an effect that was reversed by strophanthidin. The data were compared using a non-parametric repeated measures ANOVA (Friedman test) with a Dunn post-test (N neurons = 66, N slices = 10 and N mice = 5).

    Techniques Used: Concentration Assay, Mouse Assay

    The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution, and it is not recovered by α-pompilidotoxin application. a. Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 3 – Supplement 2 ), the late increase (“overshoot”) in the NAD(P)H autofluorescence and the late decrease (“undershoot”) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. b—c. Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.
    Figure Legend Snippet: The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution, and it is not recovered by α-pompilidotoxin application. a. Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 3 – Supplement 2 ), the late increase (“overshoot”) in the NAD(P)H autofluorescence and the late decrease (“undershoot”) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. b—c. Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.

    Techniques Used: Mouse Assay, Derivative Assay

    2) Product Images from "The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle"

    Article Title: The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle

    Journal: eLife

    doi: 10.7554/eLife.64821

    The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution and is not recovered by α-pompilidotoxin application. ( a ) Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 4—figure supplement 2 ), the late increase (‘overshoot’) in the NAD(P)H autofluorescence and the late decrease (‘undershoot’) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. ( b–c ) Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.
    Figure Legend Snippet: The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution and is not recovered by α-pompilidotoxin application. ( a ) Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 4—figure supplement 2 ), the late increase (‘overshoot’) in the NAD(P)H autofluorescence and the late decrease (‘undershoot’) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. ( b–c ) Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.

    Techniques Used: Mouse Assay, Derivative Assay

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    Alomone Labs α pompilidotoxin
    Neuronal stimulation triggers glycolysis in response to energy demand from ion pumping. a. Left: Representative trace of Peredox and RCaMP1h lifetimes simultaneously recorded in a DGC from an acute hippocampal slice. The slice was superfused with 2 µM GSK-2837808A for at least 30 min before the experiment, and the LDH inhibitor was kept in the ACSF during the experiment. The ACSF also contained 1 mM EGTA to reinforce Ca 2+ removal in the nominal 0Ca 2+ condition (but [Ca 2+ ] in the control ACSF was accordingly adjusted to a free concentration of 2 mM, as in any other experiment). Effective Ca 2+ removal was confirmed by the absence of a RCaMP1h spike upon stimulation. The Peredox lifetime at baseline, and the metabolic transients in response to neuronal stimulation, were recorded after substituting the bath solution with a 0Ca 2+ ACSF (to obtain Na + -only NADH CYT responses), and the further application of 10 µM <t>α-pompilidotoxin</t> (α-Pmtx, a toxin that prevents voltage-gated Na + channel inactivation). Right: The Na + -only NADH CYT transient was increased in the presence of α-pompilidotoxin. The Peredox lifetime change in response to stimulation was diminished in the absence of Ca 2+ but bounced back to higher amplitudes by increasing Na + influx. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test (N neurons = 35, N slices = 5 and N mice = 3). For all panels, only neurons showing an initial ΔPeredox lifetime response ≥0.05 ns were included for analysis. b. Representative trace of Peredox and RCaMP1h lifetimes in a DGC stimulated with trains of 100 and 200 electrical pulses. The two-stimulation protocol was also performed in 0Ca 2+ ACSF before and after the application of 3 µM α-pompilidotoxin. The latter condition was followed by the application of the Na + /K + ATPase inhibitor strophanthidin. As in (a) , the slices were exposed to the LDH inhibitor GSK-2837808A from 30 min prior, until the end of the experiment. Likewise, all the solutions contained 1 mM EGTA. c. Comparison of the Peredox lifetime changes in response to both stimulation paradigms (100 or 200 pulses) among the conditions in (b) . The NADH CYT transients in control condition was different from the other conditions (the discontinuous line for the associated p-value applies to all comparisons). The Na + -only NADH CYT responses recorded in 0Ca 2+ ACSF were increased slightly but significantly increased by the application of 3 µM α-pompilidotoxin, an effect that was reversed by strophanthidin. The data were compared using a non-parametric repeated measures ANOVA (Friedman test) with a Dunn post-test (N neurons = 66, N slices = 10 and N mice = 5).
    α Pompilidotoxin, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/α pompilidotoxin/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    α pompilidotoxin - by Bioz Stars, 2022-07
    93/100 stars
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    Neuronal stimulation triggers glycolysis in response to energy demand from ion pumping. a. Left: Representative trace of Peredox and RCaMP1h lifetimes simultaneously recorded in a DGC from an acute hippocampal slice. The slice was superfused with 2 µM GSK-2837808A for at least 30 min before the experiment, and the LDH inhibitor was kept in the ACSF during the experiment. The ACSF also contained 1 mM EGTA to reinforce Ca 2+ removal in the nominal 0Ca 2+ condition (but [Ca 2+ ] in the control ACSF was accordingly adjusted to a free concentration of 2 mM, as in any other experiment). Effective Ca 2+ removal was confirmed by the absence of a RCaMP1h spike upon stimulation. The Peredox lifetime at baseline, and the metabolic transients in response to neuronal stimulation, were recorded after substituting the bath solution with a 0Ca 2+ ACSF (to obtain Na + -only NADH CYT responses), and the further application of 10 µM α-pompilidotoxin (α-Pmtx, a toxin that prevents voltage-gated Na + channel inactivation). Right: The Na + -only NADH CYT transient was increased in the presence of α-pompilidotoxin. The Peredox lifetime change in response to stimulation was diminished in the absence of Ca 2+ but bounced back to higher amplitudes by increasing Na + influx. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test (N neurons = 35, N slices = 5 and N mice = 3). For all panels, only neurons showing an initial ΔPeredox lifetime response ≥0.05 ns were included for analysis. b. Representative trace of Peredox and RCaMP1h lifetimes in a DGC stimulated with trains of 100 and 200 electrical pulses. The two-stimulation protocol was also performed in 0Ca 2+ ACSF before and after the application of 3 µM α-pompilidotoxin. The latter condition was followed by the application of the Na + /K + ATPase inhibitor strophanthidin. As in (a) , the slices were exposed to the LDH inhibitor GSK-2837808A from 30 min prior, until the end of the experiment. Likewise, all the solutions contained 1 mM EGTA. c. Comparison of the Peredox lifetime changes in response to both stimulation paradigms (100 or 200 pulses) among the conditions in (b) . The NADH CYT transients in control condition was different from the other conditions (the discontinuous line for the associated p-value applies to all comparisons). The Na + -only NADH CYT responses recorded in 0Ca 2+ ACSF were increased slightly but significantly increased by the application of 3 µM α-pompilidotoxin, an effect that was reversed by strophanthidin. The data were compared using a non-parametric repeated measures ANOVA (Friedman test) with a Dunn post-test (N neurons = 66, N slices = 10 and N mice = 5).

    Journal: bioRxiv

    Article Title: The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle

    doi: 10.1101/2020.11.16.385526

    Figure Lengend Snippet: Neuronal stimulation triggers glycolysis in response to energy demand from ion pumping. a. Left: Representative trace of Peredox and RCaMP1h lifetimes simultaneously recorded in a DGC from an acute hippocampal slice. The slice was superfused with 2 µM GSK-2837808A for at least 30 min before the experiment, and the LDH inhibitor was kept in the ACSF during the experiment. The ACSF also contained 1 mM EGTA to reinforce Ca 2+ removal in the nominal 0Ca 2+ condition (but [Ca 2+ ] in the control ACSF was accordingly adjusted to a free concentration of 2 mM, as in any other experiment). Effective Ca 2+ removal was confirmed by the absence of a RCaMP1h spike upon stimulation. The Peredox lifetime at baseline, and the metabolic transients in response to neuronal stimulation, were recorded after substituting the bath solution with a 0Ca 2+ ACSF (to obtain Na + -only NADH CYT responses), and the further application of 10 µM α-pompilidotoxin (α-Pmtx, a toxin that prevents voltage-gated Na + channel inactivation). Right: The Na + -only NADH CYT transient was increased in the presence of α-pompilidotoxin. The Peredox lifetime change in response to stimulation was diminished in the absence of Ca 2+ but bounced back to higher amplitudes by increasing Na + influx. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test (N neurons = 35, N slices = 5 and N mice = 3). For all panels, only neurons showing an initial ΔPeredox lifetime response ≥0.05 ns were included for analysis. b. Representative trace of Peredox and RCaMP1h lifetimes in a DGC stimulated with trains of 100 and 200 electrical pulses. The two-stimulation protocol was also performed in 0Ca 2+ ACSF before and after the application of 3 µM α-pompilidotoxin. The latter condition was followed by the application of the Na + /K + ATPase inhibitor strophanthidin. As in (a) , the slices were exposed to the LDH inhibitor GSK-2837808A from 30 min prior, until the end of the experiment. Likewise, all the solutions contained 1 mM EGTA. c. Comparison of the Peredox lifetime changes in response to both stimulation paradigms (100 or 200 pulses) among the conditions in (b) . The NADH CYT transients in control condition was different from the other conditions (the discontinuous line for the associated p-value applies to all comparisons). The Na + -only NADH CYT responses recorded in 0Ca 2+ ACSF were increased slightly but significantly increased by the application of 3 µM α-pompilidotoxin, an effect that was reversed by strophanthidin. The data were compared using a non-parametric repeated measures ANOVA (Friedman test) with a Dunn post-test (N neurons = 66, N slices = 10 and N mice = 5).

    Article Snippet: Stock solutions of MgCl2 (1M) were purchased from Teknova (Hollister, CA). dorsomorphin dihydrochloride (Compound C) and GSK-2837808A were obtained from Tocris (Bristol, UK), EGTA-AM from Anaspec Inc (Fremont, CA) and α-pompilidotoxin from Alomone Labs (Jerusalem, Israel).

    Techniques: Concentration Assay, Mouse Assay

    The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution, and it is not recovered by α-pompilidotoxin application. a. Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 3 – Supplement 2 ), the late increase (“overshoot”) in the NAD(P)H autofluorescence and the late decrease (“undershoot”) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. b—c. Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.

    Journal: bioRxiv

    Article Title: The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle

    doi: 10.1101/2020.11.16.385526

    Figure Lengend Snippet: The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution, and it is not recovered by α-pompilidotoxin application. a. Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 3 – Supplement 2 ), the late increase (“overshoot”) in the NAD(P)H autofluorescence and the late decrease (“undershoot”) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. b—c. Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.

    Article Snippet: Stock solutions of MgCl2 (1M) were purchased from Teknova (Hollister, CA). dorsomorphin dihydrochloride (Compound C) and GSK-2837808A were obtained from Tocris (Bristol, UK), EGTA-AM from Anaspec Inc (Fremont, CA) and α-pompilidotoxin from Alomone Labs (Jerusalem, Israel).

    Techniques: Mouse Assay, Derivative Assay

    The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution and is not recovered by α-pompilidotoxin application. ( a ) Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 4—figure supplement 2 ), the late increase (‘overshoot’) in the NAD(P)H autofluorescence and the late decrease (‘undershoot’) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. ( b–c ) Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.

    Journal: eLife

    Article Title: The distinct roles of calcium in rapid control of neuronal glycolysis and the tricarboxylic acid cycle

    doi: 10.7554/eLife.64821

    Figure Lengend Snippet: The late overshoot in the NAD(P)H signal disappears in a Ca 2+ -free solution and is not recovered by α-pompilidotoxin application. ( a ) Left: Representative trace of the autofluorescence and RCaMP signals in the DGC layer of an acute hippocampal slice. These experiments were performed with samples from juvenile wild-type mice. When Ca 2+ was removed from the ACSF (as in Figure 4—figure supplement 2 ), the late increase (‘overshoot’) in the NAD(P)H autofluorescence and the late decrease (‘undershoot’) in the FAD + signal were almost abolished. Reducing voltage-gate Na + channel inactivation with 25 µM α-pompilidotoxin (α-Pmtx), which increases Na + influx during action potentials and results in higher energy demand due to ion pumping, did not rescue any of these signals. As expected, the application of 1 µM Tetrodotoxin (TTX), a blocker of voltage-gate Na + channels that prevents the firing of action potentials, inhibited all the signals elicited upon stimulation. Right: Average traces of the NAD(P)H and FAD + signals, expressed as the ΔF/F baseline-i ratio (mean ± SD of number of slices, N slices = 5, N mice = 3). The initial segment of the signals was zoomed in to better visualize the effects of the manipulations on the initial transient phases of both the NAD(P)H and the FAD + signals. ( b–c ) Comparisons of the parameters derived from the NAD(P)H and the FAD + signals, respectively. While Ca 2+ removal recapitulated the effects on the NAD(P)H signal, the decrease in the FAD + undershoot was more prominent than the observed in DGCs lacking MCU. The data were compared using a repeated measures ANOVA with a Student-Newman-Keuls post-test when all the groups presented Gaussian distributions, otherwise, a non-parametric Friedman test with a Dunn post-test was performed.

    Article Snippet: Stock solutions of MgCl2 (1M) were purchased from Teknova (Hollister, CA). dorsomorphin dihydrochloride (Compound C) and GSK-2837808A were obtained from Tocris (Bristol, UK), EGTA-AM from Anaspec Inc (Fremont, CA) and α-pompilidotoxin from Alomone Labs (Jerusalem, Israel).

    Techniques: Mouse Assay, Derivative Assay