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Glucose, Lactate and Glutamine but not Glutamate Support Depolarization-Induced Increased Respiration in Isolated Nerve Terminals

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Abstract

Synaptosomes prepared from various aged and gene modified experimental animals constitute a valuable model system to study pre-synaptic mechanisms. Synaptosomes were isolated from whole brain and the XFe96 extracellular flux analyzer (Seahorse Bioscience) was used to study mitochondrial respiration and glycolytic rate in presence of different substrates. Mitochondrial function was tested by sequentially exposure of the synaptosomes to the ATP synthase inhibitor, oligomycin, the uncoupler FCCP (carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone) and the electron transport chain inhibitors rotenone and antimycin A. The synaptosomes exhibited intense respiratory activity using glucose as substrate. The FCCP-dependent respiration was significantly higher with 10 mM glucose compared to 1 mM glucose. Synaptosomes also readily used pyruvate as substrate, which elevated basal respiration, activity-dependent respiration induced by veratridine and the respiratory response to uncoupling compared to that obtained with glucose as substrate. Also lactate was used as substrate by synaptosomes but in contrast to pyruvate, mitochondrial lactate mediated respiration was comparable to respiration using glucose as substrate. Synaptosomal respiration using glutamate and glutamine as substrates was significantly higher compared to basal respiration, whereas oligomycin-dependent and FCCP-induced respiration was lower compared to the responses obtained in the presence of glucose as substrate. We provide evidence that synaptosomes are able to use besides glucose and pyruvate also the substrates lactate, glutamate and glutamine to support their basal respiration. Veratridine was found to increase respiration supported by glucose, pyruvate, lactate and glutamine and FCCP was found to increase respiration supported by glucose, pyruvate and lactate. This was not the case when glutamate was the only energy substrate.

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Acknowledgments

Michaela C. Hohnholt would like to thank the “Deutsche Forschungsförderung (DFG)” for financial support (HO 5204/2-1). Helle S. Waagepetersen thanks the Carlsberg Foundation for financial support (Grant Number 108559100). The authors are grateful to Prof. Brian M. Polster for helpful discussions during the experimental study. We thank Prof. Arne Schousboe for critically reading the manuscript and many helpful suggestions. Gert H. Hansen and Lise-Lotte Niels-Christiansen are kindly acknowledged for providing the electron micrographs of synaptosomes.

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  1. Department of Drug Design and Pharmacology, Faculty of Health and Medical Science, University of Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark

    Michaela C. Hohnholt, Vibe H. Andersen, Lasse K. Bak & Helle S. Waagepetersen

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  1. Michaela C. Hohnholt
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  2. Vibe H. Andersen
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  3. Lasse K. Bak
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  4. Helle S. Waagepetersen
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Correspondence to Michaela C. Hohnholt or Helle S. Waagepetersen.

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Hohnholt, M.C., Andersen, V.H., Bak, L.K. et al. Glucose, Lactate and Glutamine but not Glutamate Support Depolarization-Induced Increased Respiration in Isolated Nerve Terminals. Neurochem Res 42, 191–201 (2017). https://doi.org/10.1007/s11064-016-2036-4

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  • DOI: https://doi.org/10.1007/s11064-016-2036-4

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