Single-Cell Imaging of Bioenergetic Responses to Neuronal Excitotoxicity and Oxygen and Glucose Deprivation
Open accessAbstract
Excitotoxicity is a condition occurring during cerebral ischemia, seizures, and chronic neurodegeneration. It is characterized by overactivation of glutamate receptors, leading to excessive Ca(2+)/Na(+) influx into neurons, energetic stress, and subsequent neuronal injury. We and others have previously investigated neuronal populations to study how bioenergetic parameters determine neuronal injury; however, such experiments are often confounded by population-based heterogeneity and the contribution of effects of non-neuronal cells. Hence, we here characterized bioenergetics during transient excitotoxicity in rat and mouse primary neurons at the single-cell level using fluorescent sensors for intracellular glucose, ATP, and activation of the energy sensor AMP-activated protein kinase (AMPK). We identified ATP depletion and recovery to energetic homeostasis, along with AMPK activation, as surprisingly rapid and plastic responses in two excitotoxic injury paradigms. We observed rapid recovery of neuronal ATP levels also in the absence of extracellular glucose, or when glycolytic ATP production was inhibited, but found mitochondria to be critical for fast and complete energetic recovery. Using an injury model of oxygen and glucose deprivation, we identified a similarly rapid bioenergetics response, yet with incomplete ATP recovery and decreased AMPK activity. Interestingly, excitotoxicity also induced an accumulation of intracellular glucose, providing an additional source of energy during and after excitotoxicity-induced energy depletion. We identified this to originate from extracellular, AMPK-dependent glucose uptake and from intracellular glucose mobilization. Surprisingly, cells recovering their elevated glucose levels faster to baseline survived longer, indicating that the plasticity of neurons to adapt to bioenergetic challenges is a key indicator of neuronal viability.
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In vitro studies have shown that neuronal ATP levels decrease only under hyperexcitation induced by strong electrical stimulation or exogenous glutamate exposure, or following pharmacological inhibition of mitochondrial ATP production pathways.
“Indeed, in vitro studies have shown that neuronal ATP levels decrease only under hyperexcitation induced by strong electrical stimulation or exogenous glutamate exposure, or following pharmacological inhibition of mitochondrial ATP production pathways20–22.”
From Energy paradox in REM sleep: balancing supply and consumption in brain metabolism (2026) · cites this paper as [21]
The ATP-depletion Results section reports a decrease during glutamate stimulation, but also during oxygen and glucose deprivation (OGD), contrary to the claim’s ‘only’ restriction. The ATP-recovery section further reports that oligomycin, an inhibitor of mitochondrial ATP production, had no measurable effect on ATP in neurons at rest; it exacerbated depletion when combined with glutamate exposure. (LLM, conf=0.88)
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Cites
AMPK: a nutrient and energy sensor that maintains energy homeostasis
Hardie, D. G., Ross, F. A., & Hawley, S. A. (2012) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3311 · Cited by 4,602 · see citing works
Bonfoco, E., Krainc, D., Ankarcrona, M., Nicotera, P., & Lipton, S. A. (1995) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.92.16.7162 · Cited by 1,958 · see citing works
Ankarcrona, M., Dypbukt, J. M., Bonfoco, E., Zhivotovsky, B., Orrenius, S., Lipton, S. A., & Nicotera, P. (1995) · Neuron · doi:10.1016/0896-6273(95)90186-8 · Cited by 1,884 · see citing works
Synaptic versus extrasynaptic NMDA receptor signalling: implications for neurodegenerative disorders
Hardingham, G. E., & Bading, H. (2010) · Nature Reviews Neuroscience · doi:10.1038/nrn2911 · Cited by 1,644 · cited 2× · see citing works
“…caused by excessive release of the excitatory neurotransmitter glutamate that contributes to the pathophysiology of neurodegenerative disorders, ischemic stroke, and traumatic brain injury (Lee et al., 1999; Wang and Qin, 2010; Hardingham and Bading, 2010; Rodriguez-Rodriguez et al., 2013).”
BackgroundRole of calcium ions in regulation of mammalian intramitochondrial metabolism
McCormack, J. G., Halestrap, A. P., & Denton, R. M. (1990) · Physiological Reviews · doi:10.1152/physrev.1990.70.2.391 · Cited by 1,398 · cited 1× · see citing works
“Indeed, cytosolic Ca 2 influx, leading to increased mitochondrial Ca 2 , has been shown to elevate TCA cycle enzymes under physiological conditions, thereby increasing mitochondrial respiration (Hansford and Castro, 1985; McCormack et al., 1990; Gunter et al., 1994).”
BackgroundMitochondria and neuronal survival
Nicholls, D. G., & Budd, S. L. (2000) · Physiological Reviews · doi:10.1152/physrev.2000.80.1.315 · Cited by 1,243 · see citing works
The changing landscape of ischaemic brain injury mechanisms
Lee, J.-M., Zipfel, G. J., & Choi, D. W. (1999) · Nature · doi:10.1038/399a007 · Cited by 1,102 · see citing works
Imamura, H., Nhat, K. P. H., Togawa, H., Saito, K., Iino, R., Kato‐Yamada, Y., Nagai, T., & Noji, H. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0904764106 · Cited by 1,089 · see citing works
Mitochondrial calcium transport: physiological and pathological relevance
Gunter, T. E., Gunter, K. K., Sheu, S. S., & Gavin, C. E. (1994) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.1994.267.2.c313 · Cited by 698 · see citing works
Goldberg, M. P., & Choi, D. W. (1993) · Journal of Neuroscience · doi:10.1523/jneurosci.13-08-03510.1993 · Cited by 693 · see citing works
The ADP and ATP transport in mitochondria and its carrier
Klingenberg, M. (2008) · Biochimica et Biophysica Acta (BBA) - Biomembranes · doi:10.1016/j.bbamem.2008.04.011 · Cited by 613 · see citing works
Molecular and cellular mechanisms of excitotoxic neuronal death
Wang, Y., & Qin, Z. (2010) · APOPTOSIS · doi:10.1007/s10495-010-0481-0 · Cited by 447 · see citing works
Mechanism suppressing glycogen synthesis in neurons and its demise in progressive myoclonus epilepsy
Vı́lchez, D., R�os, S., Cifuentes, D., Pujadas, L., Vallès, J., García-Fojeda, B., Criado‐García, O., Fernández-Sánchez, E., Medraño-Fernández, I., Domínguez, J. E., et al. (2007) · Nature Neuroscience · doi:10.1038/nn1998 · Cited by 374 · see citing works
A Reevaluation of the Role of Mitochondria in Neuronal Ca2+ Homeostasis
Budd, S. L., & Nicholls, D. G. (1996) · Journal of Neurochemistry · doi:10.1046/j.1471-4159.1996.66010403.x · Cited by 346 · see citing works
Pharmacological inhibition of AMP-activated protein kinase provides neuroprotection in stroke
McCullough, L. D., Zeng, Z., Li, H., Landree, L. E., McFadden, J., & Ronnett, G. V. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m409985200 · Cited by 333 · see citing works
Takanaga, H., Chaudhuri, B., & Frommer, W. B. (2008) · Biochimica et Biophysica Acta (BBA) - Biomembranes · doi:10.1016/j.bbamem.2007.11.015 · Cited by 333 · see citing works
Mitochondrial Membrane Potential and Glutamate Excitotoxicity in Cultured Cerebellar Granule Cells
Ward, M. W., Rego, A. C., Frenguelli, B. G., & Nicholls, D. G. (2000) · Journal of Neuroscience · doi:10.1523/jneurosci.20-19-07208.2000 · Cited by 306 · see citing works
Imaging of the Dynamics of Glucose Uptake in the Cytosol of COS-7 Cells by Fluorescent Nanosensors
Fehr, M., Lalonde, S., Lager, I., Wolff, M. W., & Frommer, W. B. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m301333200 · Cited by 295 · see citing works
Takemoto, K., Nagai, T., Miyawaki, A., & Miura, M. (2003) · The Journal of Cell Biology · doi:10.1083/jcb.200207111 · Cited by 272 · see citing works
Activation of Calpain I Converts Excitotoxic Neuron Death into a Caspase-independent Cell Death
Lankiewicz, S., Luetjens, C. M., Bui, N. T., Krohn, A. J., Poppe, M., Cole, G. M., Saido, T. C., & Prehn, J. H. M. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.275.22.17064 · Cited by 260 · see citing works
Self-citationNeurons Have an Active Glycogen Metabolism that Contributes to Tolerance to Hypoxia
Sáez, I., Durán, J., Sinadinos, C., Beltran, A., Yanes, Ó., Tevy, M. F., Martínez-Pons, C., Milán, M., & Guinovart, J. J. (2014) · Journal of Cerebral Blood Flow & Metabolism · doi:10.1038/jcbfm.2014.33 · Cited by 211 · see citing works
Mitochondrial Control of Acute Glutamate Excitotoxicity in Cultured Cerebellar Granule Cells
Castilho, R. F., Hansson, O., Ward, M. W., Budd, S. L., & Nicholls, D. G. (1998) · The Journal of Neuroscience · doi:10.1523/jneurosci.18-24-10277.1998 · Cited by 197 · see citing works
Calcium, free radicals and excitotoxins in neuronal apoptosis
Lipton, S. A., & Nicotera, P. (1998) · Cell Calcium · doi:10.1016/s0143-4160(98)90115-4 · Cited by 188 · see citing works
Nuclear translocation of AMPK-α1 potentiates striatal neurodegeneration in Huntington’s disease
Ju, T., Chen, H., Lin, J., Chang, C., Chang, W.-C., Kang, J.-J., Sun, C., Tao, M., Tu, P., Chang, C., et al. (2011) · The Journal of Cell Biology · doi:10.1083/jcb.201105010 · Cited by 183 · see citing works
Weisová, P., Concannon, C. G., Devocelle, M. C., Prehn, J. H. M., & Ward, M. W. (2009) · Journal of Neuroscience · doi:10.1523/jneurosci.0354-09.2009 · Cited by 176 · see citing works
Self-citationA Fluorescent Reporter of AMPK Activity and Cellular Energy Stress
Tsou, P., Zheng, B., Hsu, C., Sasaki, A. T., & Cantley, L. C. (2011) · Cell Metabolism · doi:10.1016/j.cmet.2011.03.006 · Cited by 158 · see citing works
Concannon, C. G., Tuffy, L. P., Weisová, P., Bonner, H. P., Dávila, D., Bonner, C., Devocelle, M. C., Strasser, A., Ward, M. W., & Prehn, J. H. M. (2010) · The Journal of Cell Biology · doi:10.1083/jcb.200909166 · Cited by 149 · see citing works
Self-citationRodriguez‐Rodriguez, P., Fernandez, E., Almeida, Á., & Bolaños, J. P. (2012) · Cell Death and Differentiation · doi:10.1038/cdd.2012.33 · Cited by 147 · see citing works
Jekabsons, M. B., & Nicholls, D. G. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m401540200 · Cited by 140 · see citing works
Ward, M. W., Huber, H. J., Weisová, P., Düßmann, H., Nicholls, D. G., & Prehn, J. H. M. (2007) · Journal of Neuroscience · doi:10.1523/jneurosci.1984-07.2007 · Cited by 114 · see citing works
Self-citationParadoxical ATP Elevation in Ischemic Penumbra Revealed by Quantitative Imaging Mass Spectrometry
Hattori, K., Kajimura, M., Hishiki, T., Nakanishi, T., Kubo, A., Nagahata, Y., Ohmura, M., Yachie‐Kinoshita, A., Matsuura, T., Morikawa, T., et al. (2010) · Antioxidants and Redox Signaling · doi:10.1089/ars.2010.3290 · Cited by 113 · see citing works
Nicholls, D. G., Vesce, S., Kirk, L., & Chalmers, S. (2003) · Cell Calcium · doi:10.1016/s0143-4160(03)00144-1 · Cited by 108 · see citing works
Role of Ca2+ in pyruvate dehydrogenase interconversion in brain mitochondria and synaptosomes
Hansford, R. G., & Castro, F. (1985) · Biochemical Journal · doi:10.1042/bj2270129 · Cited by 90 · see citing works
Excitotoxic Injury to Mitochondria Isolated from Cultured Neurons
Kushnareva, Y. E., Wiley, S. E., Ward, M. W., Andreyev, A. Y., & Murphy, A. N. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m503090200 · Cited by 76 · see citing works
Rodriguez‐Rodriguez, P., Almeida, Á., & Bolaños, J. P. (2013) · Neurochemistry International · doi:10.1016/j.neuint.2013.02.005 · Cited by 74 · see citing works
Dávila, D., Connolly, N. M. C., Bonner, H. P., Weisová, P., Düßmann, H., Concannon, C. G., Huber, H. J., & Prehn, J. H. M. (2012) · Cell Death and Differentiation · doi:10.1038/cdd.2012.49 · Cited by 74 · see citing works
Self-citationWard, M. W., Rehm, M., Düßmann, H., et al. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m511562200 · Cited by 73 · see citing works
Calpains Are Downstream Effectors ofbax-Dependent Excitotoxic Apoptosis
D’Orsi, B., Bonner, H. P., Tuffy, L. P., et al. (2012) · Journal of Neuroscience · doi:10.1523/jneurosci.2345-11.2012 · Cited by 73 · see citing works
We are all individuals: causes and consequences of non-genetic heterogeneity in mammalian cells
Loewer, A., & Lahav, G. (2011) · Current Opinion in Genetics & Development · doi:10.1016/j.gde.2011.09.010 · Cited by 70 · see citing works
Iijima, T., Mishima, T., Tohyama, M., et al. (2003) · Neurochemistry International · doi:10.1016/s0197-0186(02)00228-0 · Cited by 65 · see citing works
Marcaida, G., ̃ana, M.-D. M., Grisolı́a, S., et al. (1995) · Brain Research · doi:10.1016/0006-8993(95)00703-s · Cited by 55 · see citing works
Vergun, O. V. (2003) · Experimental Neurology · doi:10.1016/s0014-4886(03)00243-7 · Cited by 38 · see citing works
Garcı́a, O., Almeida, Á., Massieu, L., & Bolaños, J. P. (2004) · Journal of Neurochemistry · doi:10.1111/j.1471-4159.2004.02851.x · Cited by 31 · see citing works
Calcium Phosphate Transfection of DNA into Neurons in Primary Culture
Dudek, H., Ghosh, A., & Greenberg, M. E. (1998) · Current Protocols in Neuroscience · doi:10.1002/0471142301.ns0311s03 · Cited by 29 · see citing works
Pharmacological AMP-kinase activators have compartment-specific effects on cell physiology
Kodiha, M., Ho-Wo-Cheong, D., & Stochaj, U. (2011) · American Journal of Physiology-Cell Physiology · doi:10.1152/ajpcell.00309.2011 · Cited by 20 · see citing works
Dramatic effect of glycolysis inhibition on the cerebellar granule cells bioenergetics
Khodorov, B. I., Mikhailova, M. M., Bolshakov, A. P., Surin, A. M., Sorokina, E. G., Rozhnev, S. A., & Pinelis, V. G. (2012) · Biochemistry (Moscow) Supplement Series A Membrane and Cell Biology · doi:10.1134/s1990747812010060 · Cited by 8 · see citing works
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Non-Canonical Control of Neuronal Energy Status by the Na+ Pump
Baeza-Lehnert, F., Saab, A. S., Gutiérrez, R., Larenas, V., Díaz, E., Horn, M., Vargas, M., Hösli, L., Stobart, J., Hirrlinger, J., et al. (2019) · Cell Metabolism · doi:10.1016/j.cmet.2018.11.005 · Cited by 126 · see citing works
Intracellular ATP levels in mouse cortical excitatory neurons varies with sleep–wake states
Natsubori, A., Tsunematsu, T., Karashima, A., Imamura, H., Kabe, N., Trevisiol, A., Hirrlinger, J., Kodama, T., Sanagi, T., Masamoto, K., et al. (2020) · Communications Biology · doi:10.1038/s42003-020-01215-6 · Cited by 54 · see citing works
Energy paradox in REM sleep: balancing supply and consumption in brain metabolism
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- Authors
- Connolly, N. M. C., Dussmann, H., Anilkumar, U., Huber, H. J., & Prehn, J. H. M.
- Affiliations
- Royal College of Surgeons in Ireland (IE)
- KU Leuven (BE)
- Venue
- Journal of Neuroscience
- Publisher
- Society for Neuroscience
- Published
- 2014-07-30
- Volume
- 34
- Issue
- 31
- Pages
- 10192-10205
- ISSN
- 0270-6474 (ISSN-L)
- 1529-2401
- PMC
- PMC6608276
- OpenAlex
- W2083035386
- Cited by
- 62
- References
- 46
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 1 · 2023: 7 · 2022: 8 · 2021: 8 · 2020: 4 · 2019: 7 · 2018: 6 · 2017: 8 · 2016: 4 · 2015: 2 · 2014: 1
- FWCI
- 2.05
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Journal of Neuroscience (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Figshare (submittedVersion, cc-by-nc-nd)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Neuroscience › Neurology › Neurological Disorders and Treatments
- Keywords
- excitotoxicity, oxygen-glucose deprivation, neuronal bioenergetics, ATP depletion, AMPK, intracellular glucose, glycolysis, neuronal survival, single-cell imaging, fluorescent biosensor
- MeSH terms
- Animals, Animals, Newborn, Hypoxia, Calcium, Cells, Cultured, Cerebellum, Cerebral Cortex, Deoxyglucose, Energy Metabolism, Glucose, Glycine, Luminescent Proteins, Neurons, Neurotoxins, Glutamic Acid, Mice, Rats, Membrane Potential, Mitochondrial, Single-Cell Analysis, Optical Imaging
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