Journal: bioRxiv
Article Title: Differential Control of Inhibitory and Excitatory Nerve Terminal Function by Mitochondria
doi: 10.1101/2024.05.19.594864
Figure Lengend Snippet: A) Synaptic Ca 2+ imaged with synaptophysin-GCaMP6f in excitatory neurons with MCU knockdown (KD) and wild type (WT) during stimulation with 20 action potentials (AP), 20 Hz (n=9 WT, n=9 MCU KD, error bands SEM). B) Synaptic Ca 2+ in inhibitory neurons with MCU KD in comparison with WT inhibitory neurons, during 20 AP, 20 Hz stimulation (n=8 WT, n=10 MCU KD, error bands SEM). C) Quantification of change in synaptic Ca 2+ from A) and B). (n=9 excitatory WT, mean=99.2. n=9 excitatory MCU KD, mean=102.2. n=8 inhibitory WT, mean=75.01. n=10 inhibitory MCU KD, mean=193.8. Error bars SEM, n.s., p>0.99, **p=0.0014, Mann-Whitney test). D) Resting level of Ca 2+ detected with physin-GCaMP6f in excitatory and inhibitory neurons with and without MCU KD (n=9 excitatory WT, mean=114.2. n=8 excitatory MCU KD, mean=114.6. n=8 inhibitory WT, mean=101.9. n=10 inhibitory MCU KD, mean=117.2. Error bars SEM, comparisons n.s., Kruskal-Wallis test). E) Synaptic vesicle cycling traces from excitatory neurons with MCU KD or WT controls (n=13 WT, n=15 MCU KD, error bands SEM). F) Synaptic vesicle cycling fluorescence traces from MCU KD and WT inhibitory neurons (n=10 WT, n=15 MCU KD, error bands SEM). G) Quantification of response amplitude from E) and F) (n=13 WT excitatory, mean= 0.07869. n=15 MCU KD excitatory, mean=0.0666. n=10 WT inhibitory, mean=0.0572. n=15 MCU KD inhibitory, mean=0.1327. Error bars SEM. **p=0.0044, n.s. p=0.9471, Mann-Whitney test). H) Proposed pathway of activity-dependent mitochondrial Ca 2+ handling in inhibitory neurons.
Article Snippet: Mitochondrial and synaptic resting free Ca 2+ concentrations for cells expressing Mito4x-jRCaMP1b or synaptophysin-GCaMP6f were calculated by measuring the fluorescence at saturating [Ca 2+ ], F max , by applying ionomycin (Alomone Labs) at 500 μM in Tyrode’s buffer containing (in mM) 119 mM NaCl, 2.5 KCl, 4 CaCl 2 , 55 HEPES (pH 6.9).
Techniques: Knockdown, Comparison, MANN-WHITNEY, Fluorescence, Activity Assay
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