DOI: 10.5061/dryad.z8w9ghxnq
title: Mitochondrial pyruvate transport regulates presynaptic metabolism and neurotransmission

description: Glucose has long been considered the primary fuel source for the brain.
 However, glucose levels fluctuate in the brain during sleep or circuit
 activity, posing significant metabolic stress. Here, we demonstrate that
 the mammalian brain utilizes pyruvate as a fuel source, and pyruvate can
 support neuronal viability in the absence of glucose. Nerve terminals are
 sites of metabolic vulnerability, and we show that mitochondrial pyruvate
 uptake is a critical step in oxidative ATP production in hippocampal
 terminals. We find that the mitochondrial pyruvate carrier is
 post-translationally modified by lysine acetylation which in turn
 modulates mitochondrial pyruvate uptake. Our data reveal that the
 mitochondrial pyruvate carrier regulates distinct steps in
 neurotransmission, namely, the spatiotemporal pattern of synaptic vesicle
 release and the efficiency of vesicle retrieval, functions that have
 profound implications for synaptic plasticity. In summary, we identify
 pyruvate as a potent neuronal fuel and mitochondrial pyruvate uptake as a
 critical node for the metabolic control of neurotransmission in
 hippocampal terminals.
description: # Mitochondrial pyruvate transport regulates presynaptic metabolism and
 neurotransmission Anupama Tiwari, Jongyun Myeong, Arsalan Hashemiaghdam,
 Marion I. Stunault, Hao Zhang, Xianfeng Niu, Marissa A. Laramie, Jasmin
 Sponagel, Leah Shriver, Gary J. Patti, Vitaly A. Klyachko and Ghazaleh
 Ashrafi Correspondence: [ghazaleh@wustl.edu](mailto:ghazaleh@wustl.edu)
 [https://doi.org/10.5061/dryad.z8w9ghxnq](https://doi.org/10.5061/dryad.z8w9ghxnq) ## Description of method All the recorded movies from synapses expressing pHluroin. \- Tyrode_Lactate_Pyruvate 36 files: each file include multi synapse movies (37x37xtimeframe) \- Tyrode_Lactate_Pyruvate_UK5099 9 files: each file include multi synapse movies (37x37xtimeframe) Preprocessing: VGlut1-pHluorin- release event detection and localization at subpixel resolution were performed using MATLAB and the uTrack software package, which was kindly made available by Dr Gaudenz Danuser’s lab. Localization precision was determined directly from least-squares Gaussian fits of individual events. Event detection using ROI thresholding: To determine synapse locations, the stack of all 4,000 frames in each movie was summed to create a combined image, and localizations of individual synapses were defined as local peaks in the combined image using ImageJ. Whole-synapse VGlut1-pHluorin intensity was measured over a 0.95 mm diameter circle (11 pixels) region of interest (ROI) centered on each bouton. A threshold on the ROI intensity was set at 0.15 dF/F. Only ROIs with a peak amplitude greater than 2 standard deviations for the previous 1 s were accepted. Finally, instantaneous slopes of ROI fluorescence changes were calculated frame-by-frame, and detection threshold was set at 0.1 a.u./frame. An event was accepted only when all three criteria were met: the ROI intensity, slope, and 2 standard deviations criteria. ## Description of data and file structure Each experimental condition corresponds to a different folder. Inside each folder, one file (.m extension) corresponds to a different coverslip named after the recording date and Data number. Inside each .m file, there is a MATLAB cell array where each item corresponds to a movie (uint16) recorded from an individual synapse while the stimulation protocol is applied. ## Sharing/Access information please visit the following github repository, pHluorin directory: [https://github.com/JongyunMyeong85/mitochondrial-pyruvate-carrier-MPC-.git](https://github.com/JongyunMyeong85/mitochondrial-pyruvate-carrier-MPC-.git)