Abstract
In aerobic conditions, the proton-motive force drives oxidative phosphorylation (OXPHOS) and the conversion of ADP to ATP. In hypoxic environments, OXPHOS is impaired, resulting in energy shortfalls and the accumulation of protons and lactate. This results in cellular acidification, which may impact the activity and/or integrity of mitochondrial enzymes and in turn negatively impact mitochondrial respiration and thus aerobic ATP production. Naked mole-rats (NMRs) are among the most hypoxia-tolerant mammals and putatively experience intermittent hypoxia in their underground burrows. However, if and how NMR cardiac mitochondria are impacted by lactate accumulation in hypoxia is unknown. We predicted that lactate alters mitochondrial respiration in NMR cardiac muscle. To test this, we used high-resolution respirometry to measure mitochondrial respiration in permeabilized cardiac muscle fibres from NMRs exposed to 4 h of in vivo normoxia (21% O2) or hypoxia (7% O2). We found that: (1) cardiac mitochondria cannot directly oxidize lactate, but surprisingly, (2) lactate inhibits mitochondrial respiration, and (3) decreases complex IV maximum respiratory capacity. Finally, (4) in vivo hypoxic exposure decreases the magnitude of lactate-mediated inhibition of mitochondrial respiration. Taken together, our results suggest that lactate may retard electron transport system function in NMR cardiac mitochondria, particularly in normoxia, and that NMR hearts may be primed for anaerobic metabolism.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- ACR:
-
Acceptor control ratio
- CCCP:
-
Carbonyl cyanide m-chlorophenyl hydrazine
- ETS:
-
Electron transport system
- H+ :
-
Proton
- LDH:
-
Lactate dehydrogenase
- NAD:
-
Nicotinamide adenine dinucleotide
- NMR:
-
Naked mole-rat
- OXPHOS:
-
Oxidative phosphorylation
- PC:
-
Palmitoylcarnitine
- ROS:
-
Reactive oxygen species
- SUIT:
-
Substrate-uncoupler-inhibitor-titration
- TMPD:
-
Tetramethyl-p-phenylene-diamine
References
Adeva-Andany M, Lopez-Ojen M, Funcasta-Calderon R, Ameneiros-Rodriguez E, Donapetry-Garcia C, Vila-Altesor M, Rodriguez-Seijas J (2014) Comprehensive review on lactate metabolism in human health. Mitochondrion 17:76–100. https://doi.org/10.1016/j.mito.2014.05.007
Ainscow EK, Brand MD (1999) Top-down control analysis of ATP turnover, glycolysis and oxidative phosphorylation in rat hepatocytes. Eur J Biochem 263(3):671–685
Baldwin KM, Hooker AM, Herrick RE (1978) Lactate oxidative capacity in different types of muscle. Biochem Biophys Res Commun 83(1):151–157. https://doi.org/10.1016/0006-291x(78)90410-2
Bickler PE, Buck LT (2007) Hypoxia tolerance in reptiles, amphibians, and fishes: life with variable oxygen availability. Annu Rev Physiol 69:145–170. https://doi.org/10.1146/annurev.physiol.69.031905.162529
Boutilier RG (2001) Mechanisms of cell survival in hypoxia and hypothermia. J Exp Biol 204:3171–3181. https://doi.org/10.1074/jbc.273.6.3320
Boutilier RG, St-Pierre J (2002) Adaptive plasticity of skeletal muscle energetics in hibernating frogs: mitochondrial proton leak during metabolic depression. J Exp Biol 205(Pt 15):2287–2296
Brooks GA (1998) Mammalian fuel utilization during sustained exercise. Comp Biochem Physiol B Biochem Mol Biol 120(1):89–107. https://doi.org/10.1016/s0305-0491(98)00025-x
Brooks GA, Dubouchaud H, Brown M, Sicurello JP, Butz CE (1999) Role of mitochondrial lactate dehydrogenase and lactate oxidation in the intracellular lactate shuttle. Proc Natl Acad Sci USA 96(3):1129–1134. https://doi.org/10.1073/pnas.96.3.1129
Buck LT, Pamenter ME (2006) Adaptive responses of vertebrate neurons to anoxia–matching supply to demand. Respir Physiol Neurobiol 154(1–2):226–240. https://doi.org/10.1016/j.resp.2006.03.004
Buffenstein R, Amoroso V, Andziak B, Avdieiev S, Azpurua J, Barker AJ, Bennett NC, Brieno-Enriquez MA, Bronner GN, Coen C, Delaney MA, Dengler-Crish CM, Edrey YH, Faulkes CG, Frankel D, Friedlander G, Gibney PA, Gorbunova V, Hine C, Holmes MM, Jarvis JUM, Kawamura Y, Kutsukake N, Kenyon C, Khaled WT, Kikusui T, Kissil J, Lagestee S, Larson J, Lauer A, Lavrenchenko LA, Lee A, Levitt JB, Lewin GR, Lewis Hardell KN, Lin TD, Mason MJ, McCloskey D, McMahon M, Miura K, Mogi K, Narayan V, O’Connor TP, Okanoya K, O’Riain MJ, Park TJ, Place NJ, Podshivalova K, Pamenter ME, Pyott SJ, Reznick J, Ruby JG, Salmon AB, Santos-Sacchi J, Sarko DK, Seluanov A, Shepard A, Smith M, Storey KB, Tian X, Vice EN, Viltard M, Watarai A, Wywial E, Yamakawa M, Zemlemerova ED, Zions M, Smith ESJ (2022) The naked truth: a comprehensive clarification and classification of current “myths” in naked mole-rat biology. Biol Rev Camb Philos Soc 97(1):115–140. https://doi.org/10.1111/brv.12791
Cheng H, Pamenter ME (2021) Naked mole-rat brain mitochondria tolerate in vitro ischaemia. J Physiol. https://doi.org/10.1113/JP281942
Cheng H, Munro D, Huynh K, Pamenter ME (2021a) Naked mole-rat skeletal muscle mitochondria exhibit minimal functional plasticity in acute or chronic hypoxia. Comp Biochem Physiol B Biochem Mol Biol 255:110596. https://doi.org/10.1016/j.cbpb.2021.110596
Cheng H, Sebaa R, Malholtra N, Lacoste B, El Hankouri Z, Kirby A, Bennett NC, van Jaarsveld B, Hart DW, Tattersall GJ, Harper M, Pamenter ME (2021b) Naked mole-rat brown fat thermogenesis is diminished during hypoxia through a rapid decrease in UCP1. Nat Commun. https://doi.org/10.1038/s41467-021-27170-2 (In Press)
Dawson NJ, Ivy CM, Alza L, Cheek R, York JM, Chua B, Milsom WK, McCracken KG, Scott GR (2016) Mitochondrial physiology in the skeletal and cardiac muscles is altered in torrent ducks, Merganetta armata, from high altitudes in the Andes. J Exp Biol 219(Pt 23):3719–3728. https://doi.org/10.1242/jeb.142711
Descalzi G, Gao V, Steinman MQ, Suzuki A, Alberini CM (2019) Lactate from astrocytes fuels learning-induced mRNA translation in excitatory and inhibitory neurons. Commun Biol 2:247. https://doi.org/10.1038/s42003-019-0495-2
Devaux JBL, Hickey AJR, Renshaw GMC (2019) Mitochondrial plasticity in the cerebellum of two anoxia-tolerant sharks: contrasting responses to anoxia/re-oxygenation. J Exp Biol. https://doi.org/10.1242/jeb.191353
di Prampero PE (1985) Metabolic and circulatory limitations to VO2 max at the whole animal level. J Exp Biol 115:319–331
Dienel GA (2012) Brain lactate metabolism: the discoveries and the controversies. J Cereb Blood Flow Metab 32(7):1107–1138. https://doi.org/10.1038/jcbfm.2011.175
Dzal YA, Jenkin SE, Lague SL, Reichert MN, York JM, Pamenter ME (2015) Oxygen in demand: how oxygen has shaped vertebrate physiology. Comp Biochem Physiol A Mol Integr Physiol 186:4–26. https://doi.org/10.1016/j.cbpa.2014.10.029
Farhat E, Cheng H, Romestaing C, Pamenter M, Weber JM (2021) Goldfish response to chronic hypoxia mitochondrial respiration, fuel preference and energy metabolism. Metabolites. https://doi.org/10.3390/metabo11030187
Faulkes CG, Eykyn TR, Aksentijevic D (2019) Cardiac metabolomic profile of the naked mole-rat-glycogen to the rescue. Biol Let 15(11):20190710. https://doi.org/10.1098/rsbl.2019.0710
Ferguson BS, Rogatzki MJ, Goodwin ML, Kane DA, Rightmire Z, Gladden LB (2018) Lactate metabolism: historical context, prior misinterpretations, and current understanding. Eur J Appl Physiol 118(4):691–728. https://doi.org/10.1007/s00421-017-3795-6
Galli GL, Lau GY, Richards JG (2013) Beating oxygen: chronic anoxia exposure reduces mitochondrial F1FO-ATPase activity in turtle (Trachemys scripta) heart. J Exp Biol 216(Pt 17):3283–3293. https://doi.org/10.1242/jeb.087155
Gladden LB (2001) Lactic acid: new roles in a new millennium. Proc Natl Acad Sci USA 98(2):395–397. https://doi.org/10.1073/pnas.98.2.395
Gladden LB (2004) Lactate metabolism: a new paradigm for the third millennium. J Physiol 558(Pt 1):5–30. https://doi.org/10.1113/jphysiol.2003.058701
Gladden LB (2008) 200th anniversary of lactate research in muscle. Exerc Sport Sci Rev 36(3):109–115. https://doi.org/10.1097/JES.0b013e31817c0038
Gnaiger E (2014) Mitochondrial Pathways and Respiratory Control An Introduction to OXPHOS Mitochondrial Physiology Network, 29
Guppy M (2004) The biochemistry of metabolic depression: a history of perceptions. Comp Biochem Physiol B Biochem Mol Biol 139(3):435–442. https://doi.org/10.1016/j.cbpc.2004.02.019
Hashimoto T, Hussien R, Brooks GA (2006) Colocalization of MCT1, CD147, and LDH in mitochondrial inner membrane of L6 muscle cells: evidence of a mitochondrial lactate oxidation complex. Am J Physiol Endocrinol Metab 290(6):E1237-1244. https://doi.org/10.1152/ajpendo.00594.2005
Hatefi Y (1985) The mitochondrial electron transport and oxidative phosphorylation system. Annu Rev Biochem 54:1015–1069. https://doi.org/10.1146/annurev.bi.54.070185.005055
Hochachka PW (1986) Defense strategies against hypoxia and hypothermia. Science 231(4735):234–241
Hochachka PW, Buck LT, Doll CJ, Land SC (1996) Unifying theory of hypoxia tolerance: molecular/metabolic defense and rescue mechanisms for surviving oxygen lack. Proc Natl Acad Sci USA 93(18):9493–9498
Holmes MM, Goldman BD, Goldman SL, Seney ML, Forger NG (2009) Neuroendocrinology and sexual differentiation in eusocial mammals. Front Neuroendocrinol 30(4):519–533. https://doi.org/10.1016/j.yfrne.2009.04.010
Houlahan CR, Kirby AM, Dzal YA, Fairman GD, Pamenter ME (2018) Divergent behavioural responses to acute hypoxia between individuals and groups of naked mole rats. Comp Biochem Physiol B Biochem Mol Biol 224:38–44. https://doi.org/10.1016/j.cbpb.2018.01.004
Ilacqua AN, Kirby AM, Pamenter ME (2017) Behavioural responses of naked mole rats to acute hypoxia and anoxia. Biol Lett. https://doi.org/10.1098/rsbl.2017.0545
Jackson DC, Ramsey AL, Paulson JM, Crocker CE, Ultsch GR (2000) Lactic acid buffering by bone and shell in anoxic softshell and painted turtles. Physiol Biochem Zool 73(3):290–297
Jacobs RA, Meinild AK, Nordsborg NB, Lundby C (2013) Lactate oxidation in human skeletal muscle mitochondria. Am J Physiol Endocrinol Metab 304(7):E686-694. https://doi.org/10.1152/ajpendo.00476.2012
Kirby AM, Fairman GD, Pamenter ME (2018) Atypical behavioural, metabolic and thermoregulatory responses to hypoxia in the naked mole rat (Heterocephalus glaber). J Zool 305(2):106–115. https://doi.org/10.1111/jzo.12542
Kunz WS, Kuznetsov AV, Schulze W, Eichhorn K, Schild L, Striggow F, Bohnensack R, Neuhof S, Grasshoff H, Neumann HW, Gellerich FN (1993) Functional characterization of mitochondrial oxidative phosphorylation in saponin-skinned human muscle fibers. Biochim Biophys Acta 1144(1):46–53. https://doi.org/10.1016/0005-2728(93)90029-f
Lau GY, Milsom WK, Richards JG, Pamenter ME (2020) Heart mitochondria from naked mole-rats (Heterocephalus glaber) are more coupled, but similarly susceptible to anoxia-reoxygenation stress than in laboratory mice (Mus musculus). Comp Biochem Physiol B Biochem Mol Biol 240:110375. https://doi.org/10.1016/j.cbpb.2019.110375
Lin A, Krockmalnic G, Penman S (1990) Imaging cytoskeleton–mitochondrial membrane attachments by embedment-free electron microscopy of saponin-extracted cells. Proc Natl Acad Sci USA 87(21):8565–8569. https://doi.org/10.1073/pnas.87.21.8565
Mahalingam S, McClelland GB, Scott GR (2017) Evolved changes in the intracellular distribution and physiology of muscle mitochondria in high-altitude native deer mice. J Physiol 595(14):4785–4801. https://doi.org/10.1113/JP274130
Munro D, Banh S, Sotiri E, Tamanna N, Treberg JR (2016) The thioredoxin and glutathione-dependent H2O2 consumption pathways in muscle mitochondria: involvement in H2O2 metabolism and consequence to H2O2 efflux assays. Free Radic Biol Med 96:334–346. https://doi.org/10.1016/j.freeradbiomed.2016.04.014
Munro D, Baldy C, Pamenter ME, Treberg JR (2019) The exceptional longevity of the naked mole-rat may be explained by mitochondrial antioxidant defenses. Aging Cell. https://doi.org/10.1111/acel.12916
Pamenter ME, Dzal YA, Milsom WK (2015) Adenosine receptors mediate the hypoxic ventilatory response but not the hypoxic metabolic response in the naked mole rat during acute hypoxia. Proc Biol Sci 282(1800):20141722. https://doi.org/10.1098/rspb.2014.1722
Pamenter ME, Gomez CR, Richards JG, Milsom WK (2016) Mitochondrial responses to prolonged anoxia in brain of red-eared slider turtles. Biol Lett 12(1):20150797. https://doi.org/10.1098/rsbl.2015.0797
Pamenter ME, Lau GY, Richards JG (2018a) Effects of cold on murine brain mitochondrial function. PLoS ONE 13(12):e0208453. https://doi.org/10.1371/journal.pone.0208453
Pamenter ME, Lau GY, Richards JG, Milsom WK (2018b) Naked mole rat brain mitochondria electron transport system flux and H(+) leak are reduced during acute hypoxia. J Exp Biol. https://doi.org/10.1242/jeb.171397
Pamenter ME, Dzal YA, Thompson WA, Milsom WK (2019) Do naked mole rats accumulate a metabolic acidosis or an oxygen debt in severe hypoxia? J Exp Biol. https://doi.org/10.1242/jeb.191197
Pesta D, Gnaiger E (2012) High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods Mol Biol 810:25–58. https://doi.org/10.1007/978-1-61779-382-0_3
Rausch RN, Crawshaw LI, Wallace HL (2000) Effects of hypoxia, anoxia, and endogenous ethanol on thermoregulation in goldfish, Carassius auratus. Am J Physiol Regul Integr Comp Physiol 278(3):R545-555. https://doi.org/10.1152/ajpregu.2000.278.3.R545
Robergs RA, Ghiasvand F, Parker D (2004) Biochemistry of exercise-induced metabolic acidosis. Am J Physiol Regul Integr Comp Physiol 287(3):R502-516. https://doi.org/10.1152/ajpregu.00114.2004
Rogatzki MJ, Ferguson BS, Goodwin ML, Gladden LB (2015) Lactate is always the end product of glycolysis. Front Neurosci 9:22. https://doi.org/10.3389/fnins.2015.00022
Seagroves TN, Ryan HE, Lu H, Wouters BG, Knapp M, Thibault P, Laderoute K, Johnson RS (2001) Transcription factor HIF-1 is a necessary mediator of the pasteur effect in mammalian cells. Mol Cell Biol 21(10):3436–3444. https://doi.org/10.1128/MCB.21.10.3436-3444.2001
Shoubridge EA, Hochachka PW (1980) Ethanol: novel end product of vertebrate anaerobic metabolism. Science 209(4453):308–309
St-Pierre J, Brand MD, Boutilier RG (2000) Mitochondria as ATP consumers: cellular treason in anoxia. Proc Natl Acad Sci USA 97(15):8670–8674. https://doi.org/10.1073/pnas.140093597
Weibel ER (1984) The pathway for oxygen: structure and function in the mammalian respiratory system. Harvard University Press, Cambridge
Young A, Oldford C, Mailloux RJ (2020) Lactate dehydrogenase supports lactate oxidation in mitochondria isolated from different mouse tissues. Redox Biol 28:101339. https://doi.org/10.1016/j.redox.2019.101339
Acknowledgements
We would like to thank Hang Cheng for his technical support and guidance in experimental design, and the uOttawa animal care and veterinary services teams for their assistance in animal handling and husbandry.
Funding
This work was supported by an NSERC Discovery grant (#2020–07119) and a Canada Research Chair to MEP.
Author information
Authors and Affiliations
Contributions
MEP conceived of the study, wrote the manuscript, and provided material and funding support. KWH performed the experiments, analyzed the data, and edited the manuscript. All authors read and gave final approval of the published version and agree to be accountable for all content therein.
Corresponding author
Ethics declarations
Conflict of interest
We have no competing interests.
Additional information
Communicated by H.V. Carey.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Huynh, K.W., Pamenter, M.E. Lactate inhibits naked mole-rat cardiac mitochondrial respiration. J Comp Physiol B 192, 501–511 (2022). https://doi.org/10.1007/s00360-022-01430-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00360-022-01430-z