Blue bar indicates light activation. b) AMPA and NMDA receptor currents in adult (15 ... in adolescent IPN neurons that failed to respond to the application of ...
Cell Reports, Volume 20
Supplemental Information
A Cooperative Mechanism Involving Ca2+-Permeable AMPA Receptors and Retrograde Activation of GABAB Receptors in Interpeduncular Nucleus Plasticity Peter Koppensteiner, Riccardo Melani, and Ipe Ninan
Supplemental Figures
Figure S1 (related to Figure 4) vMHb-IPN glutamatergic transmission in adult and adolescent ChAT-ChR2-EYFP mice. a) Examples of EPSCs recorded at -60 and +40 mV in adult and adolescent mice. Vertical lines represent measurement of peak AMPA currents at -60 mV and NMDA currents at 50 ms from the peak current at +40 mV. Blue bar indicates light activation. b) AMPA and NMDA receptor currents in adult (15 cells/4 mice) and adolescent (16 cells/4 mice) groups. AMPA and NMDA receptor current amplitudes were not significantly different between ages. c) Adolescent mice showed significantly higher AMPA/NMDA ratios.
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Figure S2 (related to Figure 5) a) Blockade of CPARs reduces excitatory transmission in IPN neurons. Left panel shows example traces of EPSCs before and 10 min after the perfusion of NASPM (100 µM). Scale bar 100 pA/25 ms. Right panel shows EPSC amplitude before and 10 min after the perfusion of NASPM (n=9 cells/ 6 mice). b) Inclusion of EGTA (10 mM) in the electrode solution blocked activity-induced plasticity in IPN neurons. Left panel shows examples traces of EPSCs before and 20 min after the 100 Hz stimulation. Scale bar 50 pA/25 ms. Right panel shows percentage changes in EPSC amplitude before and after the 100 Hz stimulation (n=9 cells/5 mice). c) Application of 100Hz stimulation did not potentiate NMDA EPSCs recorded at +50mV in the presence of DNQX, hexamethonium, mecamylamine and bicuculline. Left panel shows example traces of NMDA EPSCs before and 20 min after the 100 Hz stimulation. Scale bar 100 pA/25 ms. Right panel shows percentage changes in EPSC amplitude before and after the 100 Hz stimulation (n=9 cells/6 mice).
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Figure S3 (related to Figure 6) Pharmacological activation of GABAB receptors enhances glutamatergic transmission in IPN neurons. a) Inclusion of GABAB agonist, baclofen, produced robust increase in EPSC amplitude in IPN neurons of adult mice. Left panel shows example traces of EPSCs evoked at an interval of 50 ms before, during and after the application of baclofen (scale bar, 10 ms/50 pA). Middle left panel shows an example of a single experiment. Middle right panel shows the average EPSC amplitude before, during and after the perfusion of baclofen (9 neurons/6 mice). Right panel shows paired pulse ratio before, during and after the application of baclofen. b) Change in EPSC amplitude in adolescent IPN neurons that responded to the application of baclofen (8 neurons out of 16 neurons from 10 mice). Left panel shows an example of a single experiment. c) EPSC amplitude in adolescent IPN neurons that failed to respond to the application of baclofen (8 neurons out of 16 neurons from 10 mice). Left panel shows an example of a single experiment. Error bars represent SEM.
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Figure S4 (related to Figure 6) Inclusion of EGTA (10 mM) in the electrode solution blocked optogenetically (10 Hz for 15 sec) -induced plasticity in the IPN neurons of VGAT-ChR2-EYFP mice. Left panel shows examples traces of EPSCs before and 20 min after the light stimulation. Scale bar 10 pA/25 ms. Right panel shows percentage changes in EPSC amplitude before and after the light stimulation (n=10 cells/7 mice).
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Figure S5 (related to Figure 7) Average freezing behavior on day 1 (habituation and fear conditioning), days 2,3,4 (fear extinction) and day 5 (test) in TA (n=21), FC (n=15) and FE (n=20) groups.
