Journal: eLife

Article Title: Attenuated dopamine signaling after aversive learning is restored by ketamine to rescue escape actions

doi: 10.7554/eLife.64041

Figure Lengend Snippet: ( a ). Schematic illustrating viral transduction strategy and two-photon Ca 2+ imaging of VTA DA neurons in acute brain slices. ( b ). Example 2PLSM image of VTA DA neurons expressing GCaMP6f. Scale bar, 20 µm. ( c ). Spontaneous Ca 2+ oscillations in one neuron with and without ketamine bath application (50 µM). Black, ACSF; blue, with ketamine. ( d ). Power spectral density of Ca 2+ transients for the neuron in ( c ). ( e ). Left, quantification of max power with and without ketamine treatment (n = 410 neurons). Right, quantification of frequency at max power. Paired two-tailed t test, ACSF vs KET, Max power, p = 0.8865, Frequency at max power, p = 0.3779. ( f ). Left, histogram showing the distribution of changes in max power with ketamine application. Right, same but for frequency at max power. n = 410 neurons. ( g ). Schematic illustrating viral transduction strategy and electrophysiological recording of VTA DA neurons in acute brain slices. ( h ). Spontaneous action potentials recorded in one neuron with and without ketamine bath application (50 µM). Black, ACSF; blue, with ketamine. ( i ). Quantification of neuronal firing rate with and without ketamine treatment (n = 9 neurons from three animals). Paired two-tailed t test, ACSF vs KET, p = 0.2561. ( j ). Spontaneous EPSCs recorded in one neuron with and without ketamine bath application (50 µM). Black, ACSF; blue, with ketamine. Holding membrane potential at –70 mV, 10 µM Gabazine in ASCF for both conditions. ( k ). Left, cumulative frequency distribution of sEPSCs amplitudes. Right, quantification of sEPSC amplitude in recorded neurons. Paired two-tailed t test, ACSF vs KET, p = 0.1958. n = 17 neurons from three animals. ( l ). Same as ( k ), but for sEPSCs inter-event intervals (IEI). Paired two-tailed t test, ACSF vs KET, p = 0.8413. ( m ). Spontaneous IPSCs recorded in one neuron with and without ketamine bath application (50 µM). Black, ACSF; blue, with ketamine. Holding membrane potential at –70 mV with high chloride internal solution, 10 µM CNQX in ASCF for both conditions. ( n ). Left, cumulative frequency distribution of sIPSCs amplitudes. Right, quantification of sIPSC amplitude in recorded neurons. Paired two-tailed t test, ACSF vs KET, p = 0.9164. n = 12 neurons from two animals. ( o ). Same as ( n ), but for sIPSCs inter-event intervals (IEI). Paired two-tailed t test, ACSF vs KET, p = 0.5675. Figure 4—source data 1. Numerical data for the graphs in .

Article Snippet: In vivo injections included intraperitoneal and subcutaneous injections of ketamine (10 mg/kg, Vedco, St. Joseph, MO) and Clozapine N-oxide (3 mg/kg in vivo, 1 μM in vitro, Sigma-Aldrich); intracranial injections of ketamine (12.5 μg in 500 nl ACSF) and Chicago Sky Blue 6B (50 μg in 500 nl ACSF, Tocris, Bristol, United Kingdom).

Techniques: Transduction, Imaging, Expressing, Two Tailed Test, Membrane

Journal: eLife

Article Title: Attenuated dopamine signaling after aversive learning is restored by ketamine to rescue escape actions

doi: 10.7554/eLife.64041

Figure Lengend Snippet: ( a ). Top, schematic for selected glutamatergic input regions to VTA. Bottom, experimental timeline. ( b ). Summary data showing the percentage of failures after local infusion of ketamine or ACSF in mPFC, and ketamine in PAG, PPTg, and VTA. Two-way ANOVA, Sidak’s multiple comparison test, LH vs LH+ KET, mPFC, p< 0.0001, PAG, p = 0.4965, PPTg, p = 0.9998, VTA, p = 0.9986. LH vs LH+ ACSF (mPFC), p = 0.9993. ( c ). Schematic for viral transduction and VTA photometry recording of Ca 2+ transients with local ketamine delivery in mPFC. ( d ). Left, average Ca 2+ transients (mean ± SEM) in response to foot shocks at the start of induction, at the end of induction, and following local ketamine infusion. Traces are aligned to shock start time (20 trials/animal, 6 animals). Right, quantification of peak Ca 2+ transient amplitude during and after foot shock stimuli across conditions. Both positive and negative values are quantified. n = 6 animals, repeated measures one-way ANOVA, Holm-Sidak’s multiple comparison test, Peak: During shock, F (1.948, 9.739) = 5.547, p = 0.0252, Induction start vs Induction end p = 0.0433, Induction end vs LH + KET, p = 0.0823. After shock, F (1.468, 7.341) = 10.05, p = 0.0105, Induction start vs Induction end, p = 0.0462, Induction end vs LH + KET, p = 0.0147. *p < 0.05, *** p < 0.001, **** p < 0.0001. Error bars reflect SEM. Figure 5—source data 1. Numerical data for the graphs in .

Article Snippet: In vivo injections included intraperitoneal and subcutaneous injections of ketamine (10 mg/kg, Vedco, St. Joseph, MO) and Clozapine N-oxide (3 mg/kg in vivo, 1 μM in vitro, Sigma-Aldrich); intracranial injections of ketamine (12.5 μg in 500 nl ACSF) and Chicago Sky Blue 6B (50 μg in 500 nl ACSF, Tocris, Bristol, United Kingdom).

Techniques: Comparison, Transduction

Journal: eLife

Article Title: Attenuated dopamine signaling after aversive learning is restored by ketamine to rescue escape actions

doi: 10.7554/eLife.64041

Figure Lengend Snippet:

Article Snippet: In vivo injections included intraperitoneal and subcutaneous injections of ketamine (10 mg/kg, Vedco, St. Joseph, MO) and Clozapine N-oxide (3 mg/kg in vivo, 1 μM in vitro, Sigma-Aldrich); intracranial injections of ketamine (12.5 μg in 500 nl ACSF) and Chicago Sky Blue 6B (50 μg in 500 nl ACSF, Tocris, Bristol, United Kingdom).

Techniques: Recombinant, Plasmid Preparation, RNAscope, Fluorescence, Multiplex Assay, Software