Monitoring ATP dynamics in electrically active white matter tracts
- Max-Planck-Institute for Experimental Medicine, Germany
- University of Zurich, Switzerland
- University of Leipzig, Germany
- Kyoto University, Japan
- Center Nanoscale Microscopy and Molecular Physiology of the Brain, Germany
Figures
Figure 1
Characterization of the expression pattern of the newly generated B6-Tg(Thy1.2-ATeam1.03YEMK)AJhi (ThyAT)-mouse line.
(A) Sagittal section of the brain highlights broad ATeam1.03YEMK expression in neurons in almost all brain regions with the exception of the olfactory bulb. Scale bar: 1 mm. (B,C) ThyAT expression …
Figure 2
Imaging of ATP combined with electrophysiology in acutely isolated optic nerves of ThyAT-mice.
(A) Binding of ATP induces a conformational change in the genetically encoded ATP-sensor ATeam1.03YEMK thus increasing the FRET effect (YFP emission upon CFP excitation) and simultaneous decreased …
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Figure 2—source data 1
Table containing data for Figure 2.
This xlsx-data file contains the data shown in Figure 2D,F and I.
- https://doi.org/10.7554/eLife.24241.005
Figure 3 with 2 supplements
Impairment of axonal ATP and CAP by glucose deprivation and/or inhibition of mitochondrial respiration.
(A) Removal of glucose from the aCSF (glucose deprivation, GD, 45 min) induces similar ATP (red) and CAP (black) decays starting at around 13 min after onset of the treatment. Red and black dashed, …
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Figure 3—source data 1
Table containing data for Figure 3.
This xlsx-data file contains the data shown in Figure 3D–H and Figure 3—Figure supplement 2.
- https://doi.org/10.7554/eLife.24241.007
Figure 3—figure supplement 1
Example of progression of CAP traces’ decay during energy deprivation.
Shown are single traces obtained under control conditions (baseline, dashed line) as well as during application of GD (A, total 45 min), MB (B, total 5 min) or MB+GD (C, total 5 min). Single traces …
Figure 3—figure supplement 2
Analysis of fluorescence changes of the ATP sensor during application of different models of energy deprivation to optic nerves.
Changes of the fluorescence signal relative to the baseline signal during glucose deprivation (A; GD) or mitochondrial blockage (B; MB). Fluorescence intensities of the three recorded channels …
Figure 4 with 5 supplements
Comparison of ATP and CAP dynamics during high frequency stimulation.
(A) The CAP area decreases over time during high-frequency stimulation (HFS). The decay amplitude deviates from the absence of HFS, indicated by the dashed line (0.1 Hz, used for normalization to …
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Figure 4—source data 1
Table containing data for Figure 4.
This xlsx-data file contains the data shown in Figure 4C,D,F and Figure 4—Figure supplement 5.
- https://doi.org/10.7554/eLife.24241.011
Figure 4—figure supplement 1
Example of progression of CAP traces’ decay during high frequency stimulation (HFS).
(A) The three peaks recognizable in the baseline trace (dashed line) are differently affected by increasing stimulation frequency and for CAP analysis only the first two are considered. Shown are …
Figure 4—figure supplement 2
Stimulation of optic nerves with progressively increasing frequencies.
Frequency-dependent changes in relative signal amplitude of ATP and CAP, during progressively increasing stimulation frequencies (1 Hz to 100 Hz) and following recovery. Nerves incubated in aCSF …
Figure 4—figure supplement 3
Correlation of the rates and amplitudes of CAP and ATP changes during HFS in different glucose concentrations.
(A) Correlation of the velocity of the initial decay of CAP at the beginning of HFS and the amplitude of CAP decay at the end of HFS. The faster the CAP drops, the larger the CAP amplitude is. (B) …
Figure 4—figure supplement 4
Example of CAP traces before and after high-frequency stimulation (HFS) of optic nerves incubated in aCSF with different concentrations of glucose.
Shown are mean CAP wave forms (n = 3 nerves for each condition) incubated in aCSF containing 10 mM glucose (A), 3.3 mM glucose (B) and 2 mM glucose (C) prior to high- frequency stimulation …
Figure 4—figure supplement 5
Analysis of fluorescence changes of the ATP sensor during high-frequency stimulation (HFS) of optic nerves incubated in aCSF with different concentrations of glucose.
Changes of the fluorescence signal at the end of the 2.5 min HFS (100 Hz) period relative to the baseline signal prior to stimulation of optic nerves incubated in aCSF containing 10 mM glucose (A), …
Figure 5 with 2 supplements
Energy metabolism of optic nerves depends on the type and concentration of substrates and involves lactate metabolism.
(A) The comparison between CAP area decay of optic nerves incubated in 10 mM glucose aCSF (n = 5 nerves) versus optic nerves incubated either in 10 mM lactate or 10 mM pyruvate (n = 3 nerves) during …
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Figure 5—source data 1
Table containing data for Figure 5.
This xlsx-data file contains the data shown in Figure 5A–D,F and Figure 5—Figure supplement 1B.
- https://doi.org/10.7554/eLife.24241.018
Figure 5—figure supplement 1
Correlation of the amplitudes of CAP and ATP changes in the presence of lactate and pyruvate as exogenous energy substrates.
(A) Correlation of ATP and CAP decay amplitude during HFS of nerves bathed in aCSF containing glucose, lactate or pyruvate (each 10 mM) as energy substrates. n = 5, 3, 3 nerves for glucose, lactate …
Figure 5—figure supplement 2
Example of CAP traces before and after high-frequency stimulation (HFS) of optic nerves in the presence of inhibitors of lactate metabolism.
Optic nerves were incubated in aCSF containing 3.3 mM glucose in the absence of inhibitors (A) or in the presence of either 20 mM D-lactate (B) or 10 µM AR-C155858 (C). Shown are mean CAP wave forms …
