|Gnaiger E, Aasander Frostner E, Abdul Karim N, Abumrad NA, Acuna-Castroviejo D, Adiele RC, et al (2019) Mitochondrial respiratory states and rates. MitoFit Preprint Arch doi:10.26124/mitofit:190001.v2.|
Version 2: Posted online 2019-03-15 Open Access Freely available online doi:10.26124/mitofit:190001.v2
Version 1: Posted online 2019-02-12 Open Access Freely available online doi:10.26124/mitofit:190001
Author affiliations in hyperlinks:, Gnaiger E, Aasander Frostner E, Abdul Karim N, Abumrad NA, Acuna-Castroviejo D, Adiele RC, Ahn B, Ali SS, Alton L, Alves MG, Amati F, Amoedo ND, Andreadou I, Arago M, Aral C, Arandarcikaite O, Armand AS, Arnould T, Avram VF, Bailey DM, Bajpeyi S, Bajzikova M, Bakker BM, Barlow J, Bastos Sant'Anna Silva AC, Batterson P, Battino M, Bazil J, Beard DA, Bednarczyk P, Bello F, Ben-Shachar D, Bergdahl A, Berge RK, Bergmeister L, Bernardi P, Berridge MV, Bettinazzi S, Bishop D, Blier PU, Blindheim DF, Boardman NT, Boetker HE, Borchard S, Boros M, Borsheim E, Borutaite V, Botella J, Bouillaud F, Bouitbir J, Boushel RC, Bovard J, Breton S, Brown DA, Brown GC, Brown RA, Brozinick JT, Buettner GR, Burtscher J, Calabria E, Calbet JA, Calzia E, Cannon DT, Cano Sanchez M, Canto AC, Cardoso LHD, Carvalho E, Casado Pinna M, Cassar S, Cassina AM, Castelo MP, Castro L, Cavalcanti-de-Albuquerque JP, Cervinkova Z, Chabi B, Chakrabarti L, Chakrabarti S, Chaurasia B, Chen Q, Chicco AJ, Chinopoulos C, Chowdhury SK, Cizmarova B, Clementi E, Coen PM, Cohen BH, Coker RH, Collin A, Crisostomo L, Dahdah N, Dalgaard LT, Dambrova M, Danhelovska T, Darveau CA, Das AM, Dash RK, Davidova E, Davis MS, De Goede P, De Palma C, Dembinska-Kiec A, Detraux D, Devaux Y, Di Marcello M, Dias TR, Distefano G, Doermann N, Doerrier C, Dong L, Donnelly C, Drahota Z, Duarte FV, Dubouchaud H, Duchen MR, Dumas JF, Durham WJ, Dymkowska D, Dyrstad SE, Dyson A, Dzialowski EM, Eaton S, Ehinger J, Elmer E, Endlicher R, Engin AB, Escames G, Ezrova Z, Falk MJ, Fell DA, Ferdinandy P, Ferko M, Ferreira JCB, Ferreira R, Ferri A, Fessel JP, Filipovska A, Fisar Z, Fischer C, Fischer M, Fisher G, Fisher JJ, Ford E, Fornaro M, Galina A, Galkin A, Gallee L, Galli GL, Gama Perez P, Gan Z, Ganetzky R, Garcia-Rivas G, Garcia-Roves PM, Garcia-Souza LF, Garipi E, Garlid KD, Garrabou G, Garten A, Gastaldelli A, Gayen J, Genders AJ, Genova ML, Giovarelli M, Goncalo Teixeira da Silva R, Goncalves DF, Gonzalez-Armenta JL, Gonzalez-Freire M, Gonzalo H, Goodpaster BH, Gorr TA, Gourlay CW, Granata C, Grefte S, Guarch ME, Gueguen N, Gumeni S, Haas CB, Haavik J, Haendeler J, Haider M, Hamann A, Han J, Han WH, Hancock CR, Hand SC, Handl J, Hargreaves IP, Harper ME, Harrison DK, Hassan H, Hausenloy DJ, Heales SJR, Heiestad C, Hellgren KT, Hepple RT, Hernansanz-Agustin P, Hewakapuge S, Hickey AJ, Ho DH, Hoehn KL, Hoel F, Holland OJ, Holloway GP, Hoppel CL, Hoppel F, Houstek J, Huete-Ortega M, Hyrossova P, Iglesias-Gonzalez J, Irving BA, Isola R, Iyer S, Jackson CB, Jadiya P, Jana PF, Jang DH, Jang YC, Janowska J, Jansen K, Jansen-Duerr P, Jansone B, Jarmuszkiewicz W, Jaskiewicz A, Jedlicka J, Jespersen NR, Jha RK, Jurczak MJ, Jurk D, Kaambre T, Kaczor JJ, Kainulainen H, Kampa RP, Kandel SM, Kane DA, Kapferer W, Kappler L, Karabatsiakis A, Karavaeva I, Karkucinska-Wieckowska A, Kaur S, Keijer J, Keller MA, Keppner G, Khamoui AV, Kidere D, Kilbaugh T, Kim HK, Kim JKS, Klepinin A, Klepinina L, Klingenspor M, Klocker H, Komlodi T, Koopman WJH, Kopitar-Jerala N, Kowaltowski AJ, Kozlov AV, Krajcova A, Krako Jakovljevic N, Kristal BS, Krycer JR, Kuang J, Kucera O, Kuka J, Kwak HB, Kwast K, Laasmaa M, Labieniec-Watala M, Lagarrigue S, Lai N, Land JM, Lane N, Laner V, Lanza IR, Laranjinha J, Larsen TS, Lavery GG, Lazou A, Lee HK, Leeuwenburgh C, Lehti M, Lemieux H, Lenaz G, Lerfall J, Li PA, Li Puma L, Liepins E, Liu J, Lopez LC, Lucchinetti E, Ma T, Macedo MP, Maciej S, MacMillan-Crow LA, Majtnerova P, Makarova E, Makrecka-Kuka M, Malik AN, Markova M, Martin DS, Martins AD, Martins JD, Maseko TE, Maull F, Mazat JP, McKenna HT, McKenzie M, Menze MA, Merz T, Meszaros AT, Methner A, Michalak S, Moellering DR, Moisoi N, Molina AJA, Montaigne D, Moore AL, Moreau K, Moreira BP, Moreno-Sanchez R, Mracek T, Muccini AM, Munro D, Muntane J, Muntean DM, Murray AJ, Musiol E, Nabben M, Nair KS, Nehlin JO, Nemec M, Neufer PD, Neuzil J, Neviere R, Newsom SA, Nozickova K, O'Brien KA, O'Gorman D, Olgar Y, Oliveira B, Oliveira MF, Oliveira MT, Oliveira PF, Oliveira PJ, Orynbayeva Z, Osiewacz HD, Pak YK, Pallotta ML, Palmeira CM, Parajuli N, Passos JF, Passrugger M, Patel HH, Pavlova N, Pecina P, Pedersen TM, Pereira da Silva Grilo da Silva F, Pereira SP, Perez Valencia JA, Perks KL, Pesta D, Petit PX, Pettersen IKN, Pichaud N, Pichler I, Piel S, Pietka TA, Pino MF, Pirkmajer S, Plangger M, Porter C, Porter RK, Procaccio V, Prochownik EV, Prola A, Pulinilkunnil T, Puskarich MA, Puurand M, Radenkovic F, Ramzan R, Rattan SIS, Reboredo P, Renner-Sattler K, Rial E, Robinson MM, Roden M, Rodriguez E, Rodriguez-Enriquez S, Roesland GV, Rohlena J, Rolo AP, Ropelle ER, Rossignol R, Rossiter HB, Rubelj I, Rybacka-Mossakowska J, Saada A, Safaei Z, Saharnaz S, Salin K, Salvadego D, Sandi C, Saner N, Sanz A, Sazanov LA, Scatena R, Schartner M, Scheibye-Knudsen M, Schilling JM, Schlattner U, Schoenfeld P, Schots PC, Schulz R, Schwarzer C, Scott GR, Selman C, Shabalina IG, Sharma P, Sharma V, Shevchuk I, Shirazi R, Shiroma JG, Siewiera K, Silber AM, Silva AM, Sims CA, Singer D, Singh BK, Skolik R, Smenes BT, Smith J, Soares FAA, Sobotka O, Sokolova I, Sonkar VK, Sowton AP, Sparagna GC, Sparks LM, Spinazzi M, Stankova P, Starr J, Stary C, Stelfa G, Stepto NK, Stiban J, Stier A, Stocker R, Storder J, Sumbalova Z, Suomalainen A, Suravajhala P, Svalbe B, Swerdlow RH, Swiniuch D, Szabo I, Szewczyk A, Szibor M, Tanaka M, Tandler B, Tarnopolsky MA, Tausan D, Tavernarakis N, Tepp K, Thakkar H, Thapa M, Thyfault JP, Tomar D, Ton R, Torp MK, Towheed A, Tretter L, Trewin AJ, Trifunovic A, Trivigno C, Tronstad KJ, Trougakos IP, Truu L, Tuncay E, Turan B, Tyrrell DJ, Urban T, Valentine JM, Van Bergen NJ, Van Hove J, Varricchio F, Vella J, Vendelin M, Vercesi AE, Victor VM, Vieira Ligo Teixeira C, Vidimce J, Viel C, Vieyra A, Vilks K, Villena JA, Vincent V, Vinogradov AD, Viscomi C, Vitorino RMP, Vogt S, Volani C, Volska K, Votion DM, Vujacic-Mirski K, Wagner BA, Ward ML, Warnsmann V, Wasserman DH, Watala C, Wei YH, Whitfield J, Wickert A, Wieckowski MR, Wiesner RJ, Williams CM, Winwood-Smith H, Wohlgemuth SE, Wohlwend M, Wolff JN, Wrutniak-Cabello C, Wuest RCI, Yokota T, Zablocki K, Zanon A, Zanou N, Zaugg K, Zaugg M, Zdrazilova L, Zhang Y, Zhang YZ, Zikova A, Zischka H, Zorzano A, Zvejniece L (2019) MitoFit Preprint Arch
Abstract: As the knowledge base and importance of mitochondrial physiology to human health expands, the necessity for harmonizing the terminology concerning mitochondrial respiratory states and rates has become increasingly apparent. The chemiosmotic theory establishes the mechanism of energy transformation and coupling in oxidative phosphorylation. The unifying concept of the protonmotive force provides the framework for developing a consistent theoretical foundation of mitochondrial physiology and bioenergetics. We follow guidelines of the International Union of Pure and Applied Chemistry (IUPAC) on terminology in physical chemistry, extended by considerations of open systems and thermodynamics of irreversible processes. The concept-driven constructive terminology incorporates the meaning of each quantity and aligns concepts and symbols to the nomenclature of classical bioenergetics. We endeavour to provide a balanced view on mitochondrial respiratory control and a critical discussion on reporting data of mitochondrial respiration in terms of metabolic flows and fluxes. Uniform standards for evaluation of respiratory states and rates will ultimately contribute to reproducibility between laboratories and thus support the development of databases of mitochondrial respiratory function in species, tissues, and cells. Clarity of concept and consistency of nomenclature facilitate effective transdisciplinary communication, education, and ultimately further discovery.
• Keywords: Mitochondrial respiratory control, coupling control, mitochondrial preparations, protonmotive force, uncoupling, oxidative phosphorylation, OXPHOS, efficiency, electron transfer, ET; proton leak, LEAK, residual oxygen consumption, ROX, State 2, State 3, State 4, normalization, flow, flux, O2 • Bioblast editor: Gnaiger E
Authors: MitoEAGLE Task Group
- Corresponding author
- Erich Gnaiger
- Chair COST Action CA15203 MitoEAGLE
- T +43 512 566796 15, F +43 512 566796 20
- firstname.lastname@example.org | www.mitoeagle.org
- Coauthors (listed in alphabetical order); number of coauthors (2019-03-15): 533
- Many coauthors have made significant additions and suggestions for improvement of the manuscript. All coauthors confirm to have read the final manuscript, and to agree to implement the recommendations into future manuscripts, presentations and teaching materials.
- Disclaimer: This article was prepared while Joshua P. Fessel was employed at Vanderbilt University Medical Center. The opinions expressed in this article are the author's own and do not reflect the view of the National Institutes of Health, the Department of Health and Human Services, or the United States government.
- Copyright: © 2019 Gnaiger et al.
- This is an Open Access preprint (not peer-reviewed) distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are credited. © remains with the authors, who have granted MitoFit an Open Access preprint licence in perpetuity.
Preprints for Gentle Science
- Fantastic initiative with the new mitochondrial physiology preprint server! ~ Donnelly C
- It is quite surprising that the final manuscript was not accepted in BioRvix. I completely agree with MitoFit preprint archive. ~ Singh BK
- It will be a pleasure to join the MitoEagle task group publication. ~ Laranjinha J
- I have a small remark : the concept of multiple authors signature started to be contested... By all ETHICS commities everywhere in Europa at least since it overpassed the usual rules... And also altered the signification of the authors impact factor. The regulation will be to form a consortium that is the true entity that will sign the collective work. That is teh best for teh COST since all members are easily identifiables. Overall conmment: This paper has been stanfding to long.... on teh bench! Petit PX
- I just found a minor typo. If you look at the “S” references, they are out of alphabetical order. ~ Sparagna GC
- Please find attached manuscript with comments (I have made 5 in total). Feel free to incorporate (or ignore) as you see fit! ~ McKenzie M
- Gnaiger E: To address your comment “Interesting that you state saturating O2, as this is only at the start of an experiment? (but is accounted for in the oxygraph calibration with dithionite, so that measured respiration rates are relative to saturating O2?).”, I extended Section 2.1.2: “Kinetically-saturated conditions are evaluated by substrate kinetics to obtain the maximum reaction velocity or maximum pathway flux, in contrast to solubility-saturated conditions.”
- We are in direct contact with Kyle Hoehn to obtain and test their uncouplers.
- To summarize your comment “So would it be optimal in publications to not only state final flux rates/unit sample (e.g per mg) but also the raw flux rates (per mL) and the mg of sample used?”, I extended Box 3:Box 3: Recommendations for studies with mitochondrial preparations
- ● Normalization of respiratory rates should be provided as far as possible:
- A. Sample normalization
- 1. Object-specific biophysical normalization: on a per organism or per cell basis as O2 flow; this may not be possible when dealing with coenocytic organisms, e.g., filamentous fungi, or tissues without cross-walls separating individual cells, e.g., muscle fibers.
- 2. Size-specific cellular normalization: per g protein; per organism-, cell- or tissue-mass as mass-specific O2 flux; per cell volume as cell volume-specific flux.
- 3. Mitochondrial normalization: per mitochondrial marker as mt-specific flux.
- B. Chamber normalization
- 1. Chamber volume-specific flux, JV [pmol∙s-1∙mL-1], is reported for quality control in relation to instrumental sensitivity and limit of detection of volume-specific flux.
- 2. Sample concentration in the instrumental chamber is reported as number concentration, mass concentration, or mitochondrial concentration; this is a component of the measuring conditions.
- A. Sample normalization
- With information on cell size and the use of multiple normalizations, maximum potential information is available (Renner et al. 2003; Wagner et al. 2011; Gnaiger 2014). Reporting flow in a respiratory chamber [nmol∙s-1] is discouraged, since it restricts the analysis to intra-experimental comparison of relative (qualitative) differences.
- I am happy to see that we are one step closer to the final publication of the MitoEAGLE manuscript in a journal. ~ Komlodi T
- It is a great step towards the publication of the manuscript and congratulations for creating your own tool to circumvent decisions that can not be easily understood. ~ Thierry A
- Excellent article and one of its kind too. Please let me how can i help to get it published in a high impact j. Look forward to work in your team. ~ Sharma P
- It's great to see the preprint. The preprint server in the area of mitochondrial physiology is a great idea and definitely will be a success. ~ Tomar D
- First and foremost I would like to express my deepest gratitude and would like to thank you for giving us your time to review our manuscript and be part of us as the co-author. It is a great honour to get you in touch and reply promptly. .. I would like to also thank you for giving me the opportunity to be part of the MitoEAGLE as one of the co-authors and I am happy to be listed in the next version of the preprint. ~ Hassan H
- However - just a note about pre-prints. A significant portion of scientists that I collaborate with feel uncomfortable submitting manuscript on a pre-print server. Is this something that could be addressed maybe in an article regarding the benefits and nuances of pre-print server submission. ~ Towheed A
- FAQ - MitoPedia:Preprint
Journal submission comments
- Cell Metabolism seems like a good first choice. ~ Williams C
- Cell Metabolism, I think this is a good choice. ~ Rossiter HB
- Cell Metabolism seems highly appropriate. ~ Newsom SA
- No preference. Just go ahead. ~ Zaugg M
- I concur with the choice of cell metabolism. ~ Pulinilkunnil T
- I think cell metabolism would be great, but I doubt whether it is realistic. Possibly Molecular Metabolism (very rapid, good reputation, european), Cell and Molecular Life Sciences (many reviews) or BBA- bioenergetics could be alternatives. ~ Keijer J
- Alternatives if Cell Met is not accepting: Nature metabolism or Acta physiologica. ~ Amati F
- And Cell met is a good 1st choice for this publication. ~ Zanou N
- The question is why did Biorxiv reject the manuscript? Before submitting to a prestigious journal like Cell Metabolism all the doubts Biorxiv had should be ruled out. ~ Methner A
- And I think that Cell Metabolism is a good first journal choice for submission of our manuscript. ~ Breton S
- Cell Metabolism is a good option as a first submission. ~ Bouitbir J
- Cell metabolism is a good fit for the manuscript. ~ Adiele RC
- The choice of journal is excellent, although it might be a long shot. ~ Oliveira MT
- I guess that also TIBs or Current Biology could be considered. ~ Calabria E
- I am in agreement that the first journal will be Cell Metabolism. ~ Victor VM
- Cell metabolism is an excellent choice. If they are interested that would be wonderfull. Physiological Reviews could be an alternative, in case Cell metabolism declines the manuscript. ~ Thierry A
- I would suggest to try the submission in Cell Metabolism. ~ Doerrier C
- Regarding the future submission of our paper, if there is already a pre-acceptance of the Editor of Cell Metabolism, I believe we should submit there. There are not many journals willing to publish a paper with so many authors, and the reviewing process will not be easy, in my opinion. ~ Crisostomo L
- I think cell metabolism is a good target to submit our article. ~ Salin K
- I would format accurately as a resource manuscript for Cell Metabolism. ~ Lavery GG
- The manuscript is still extremely long. In my modest opinion too long compared to the editorial format limits of many journals. If the manuscript cannot be substantially shortened to the essentials (in my opinion preferable) one strategy is to try to find a journal without such limits. ~ Spinazzi M
- I am happy with Cell Metabolism to start the submission process of this preprint. ~ Moisoi N
Linking COST Actions and MiPsociety
Towards the preprint
- » Manuscript phases and versions towards the Preprint
Manuscript phases and versions - an open-access apporach
- This manuscript on ‘Mitochondrial respiratory states and rates’ is a position statement in the frame of COST Action CA15203 MitoEAGLE. The list of coauthors evolved beyond phase 1 in the bottom-up spirit of COST.
- The global MitoEAGLE network made it possible to collaborate with a large number of coauthors to reach consensus on the present manuscript. Nevertheless, we do not consider scientific progress to be supported by ‘declaration’ statements (other than on ethical or political issues). Our manuscript aims at providing arguments for further debate rather than pushing opinions. We hope to initiate a much broader process of discussion and want to raise the awareness on the importance of a consistent terminology for reporting of scientific data in the field of bioenergetics, mitochondrial physiology and pathology. Quality of research requires quality of communication. Some established researchers in the field may not want to re-consider the use of jargon which has become established despite deficiencies of accuracy and meaning. In the long run, superior standards will become accepted. We hope to contribute to this evolutionary process, with an emphasis on harmonization rather than standardization.
- Phase 1: The protonmotive force and respiratory control
- » The protonmotive force and respiratory control - Discussion
- » MitoEAGLE preprint 2017-09-21 - Discussion
- 2017-11-11: Print version (16) for MiP2017/MitoEAGLE Hradec Kralove CZ
- Phase 2: Mitochondrial respiratory states and rates: Building blocks of mitochondrial physiology Part 1
- Phase 4: Journal submission
- Target: CELL METABOLISM, aiming at indexing by The Web of Science and PubMed.
- 2017-09-21 Version 01: 105 coauthors
- 2017-10-15 Version 10: 131 coauthors
- 2018-01-18 Version 20: 168 coauthors
- 2018-02-26 Version 30: 225 coauthors
- 2018-08-20 Version 40: 350 coauthors - EBEC Poster
- 2018-10-17 Version 44: 426 coauthors - MiPschool Tromso-Bergen 2018
- 2018-12-12 Version 50: 517 coauthors - Submission to the preprint server bioRxiv not successful
- 2019-02-12 Preprint version 1: 530 coauthors - Preprint publication doi:10.26124/mitofit:190001
- 2019-03-15 Preprint version 2: 533 coauthors - Preprint publication doi:10.26124/mitofit:190001.v2
Labels: MiParea: Respiration, mt-Awareness
Preparation: Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria Enzyme: Marker enzyme Regulation: Coupling efficiency;uncoupling, Flux control, mt-Membrane potential, Uncoupler Coupling state: LEAK, OXPHOS, ET Pathway: F, N, S, Gp, DQ, CIV, NS, Other combinations, ROX