AMPAR/TARP stoichiometry differentially modulates channel properties
- Laboratori de Neurofisiologia, Departament de Biomedicina, Facultat de Medicina i Ciències de la Salut, Institut de Neurociències, Universitat de Barcelona, Spain
- Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Spain
Figures
Figure 1
CP-AMPAR kinetics are differentially affected by AMPAR-TARP stoichiometry.
(A) Traces evoked at −60 mV by rapid application of 10 mM glutamate to outside-out patches from cells expressing GluA1 alone (black; average of 39 responses) or together with 2 (blue; average of 37 …
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Figure 1—source data 1
Kinetic properties of CP-AMPARs.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig1-data1-v3.xlsx
Figure 2
Recovery from desensitization of CP-AMPARs is enhanced in a graded manner with increased γ2.
(A) Representative traces of a two-pulse protocol with increasing time interval between pulses for CP-AMPAR without γ2 TARP (GluA1 homomers; black), with 2 γ2 TARPs (blue) and with 4 γ2 TARPs (red). …
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Figure 2—source data 1
Recovery from desensitization of CP-AMPARs.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig2-data1-v3.xlsx
Figure 3
CP-AMPAR polyamine block attenuation is dependent on TARP dosage.
(A) Representative glutamate-evoked currents from outside-out patches at different membrane potentials from −80 to +80 in 20 mV increments from cells expressing CP-AMPARs, GluA1 (black), …
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Figure 3—source data 1
Polyamine block attenuation of CP-AMPARs.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig3-data1-v3.xlsx
Figure 4 with 1 supplement
Four TARPs are required to increase CP-AMPAR channel conductance.
(A) Typical responses at a holding potential of −60 mV to rapid application of 10 mM glutamate to excised patches from cells expressing homomeric GluA1 alone (black; average of 84 responses) or …
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Figure 4—source data 1
Channel conductance of CP-AMPARs.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig4-data1-v3.xlsx.xlsx
Figure 4—figure supplement 1
Short and long isoforms of GluA4 show the same electrophysiological behaviour.
(A) Pooled data showing single channel conductance from GluA4 (light grey) and GluA4c (dark grey) homotetramers. Box-and-whiskers plots represent percentiles, median and average as stated in Figure …
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Figure 4—figure supplement 1—source data 1
Source data.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig4-figsupp1-data1-v3.xlsx
Figure 5
Single channel conductance of CI-AMPARs is modulated differently by TARPs depending on their location within the complex.
(A) Evoked currents by rapid application of 10 mM glutamate from membrane patches at +60 mV (upward traces) and −60 mV (downward traces) with their corresponding RI. The experimental conditions are …
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Figure 5—source data 1
KInetic properties of CI-AMPARs.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig5-data1-v3.xlsx
Figure 6 with 2 supplements
CI-AMPAR kinetics differ upon γ2 attachment to GluA4c or GluA2 subunit.
(A) Representative traces of currents at −60 mV from cells expressing CI-AMPARs without or with TARP γ2 linked to GluA subunits. Under the traces a scheme of the subunits forming the receptors with …
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Figure 6—source data 1
Channel conductance of CI-AMPARs.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig6-data1-v3.xlsx
Figure 6—figure supplement 1
The block of the non-competitive antagonist perampanel varies with AMPAR-TARP stoichiometry.
(A). Representative whole-cell recording showing the protocol used to determine the percentage of perampanel block. This trace corresponds to a cell expressing GluA2 and GluA4c. Cells were clamped …
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Figure 6—figure supplement 1—source data 1
Source data.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig6-figsupp1-data1-v3.xlsx
Figure 6—figure supplement 2
Effect of C549L and C550L AMPAR mutations in AMPAR-TARP modulation.
(A–B) Pooled data showing C550L mutation effect in GluA4c homotetramers in decreasing the weighted time constant of desensitization (τw, des) and single channel conductance. (C–D) C549L and C550L …
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Figure 6—figure supplement 2—source data 1
Source data.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig6-figsupp2-data1-v3.xlsx
Figure 7 with 1 supplement
Somatic currents from CGCs exhibit properties of GluA2:γ2 + GluA4c CI-AMPARs.
(A) CGCs in culture after 7 days in vitro. (B) Traces at +60 mV and −60 mV evoked with 100 μM AMPA from a CGC somatic patch showing the typical lineal response of a CI-AMPAR. (C) Representative …
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Figure 7—source data 1
Cerebellar granule cells properties.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig7-data1-v3.xlsx
Figure 7—figure supplement 1
Recovery of desensitization kinetics showing no differences between agonists used to evoke currents in 2T(A2) condition.
Data from this figure containing statistical tests applied, exact sample number, p values and details of replicates is available in ‘Figure 7—figure supplement 1—source data 1’.
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Figure 7—figure supplement 1—source data 1
Source data.
- https://cdn.elifesciences.org/articles/53946/elife-53946-fig7-figsupp1-data1-v3.xlsx
Author response image 1
