Vernerova 2021 MitoFit PLT

From Bioblast
Jump to navigation Jump to search


MitoFit Preprints         MitoFit Preprints        
Gnaiger 2019 MitoFit Preprints
       
Gnaiger MitoFit Preprints 2020.4
        MitoFit DOI Data Center         MitoPedia: Preprints


Vernerova 2021 MitoFit PLT

Publications in the MiPMap
Vernerova A, Garcia-Souza LF, Soucek O, Kostal M, Rehacek V, Krcmova LK, Gnaiger E, Sobotka O (2021) Mitochondrial respiration of platelets: comparison of isolation methods. MitoFit Preprints 2021.6. doi:10.26124/mitofit:2021-0006

»

MitoFit pdf

Mitochondrial respiration of platelets: comparison of isolation methods

Vernerova Andrea, Garcia-Souza Luiz F, Soucek Ondrej, Kostal Milan, Rehacek Vit, Krcmova Lenka K, Gnaiger Erich, Sobotka Ondrej (2021-09-16) MitoFit Prep

Abstract: Multiple non-aggregatory functions of human platelets (PLT) are widely acknowledged, yet their functional examination is limited mainly due to a lack of standardized isolation and analytic methods. Platelet apheresis (PA) is an established clinical method for PLT isolation aiming at the treatment of bleeding diathesis in severe thrombocytopenia. On the other hand, density gradient centrifugation (DC) is an isolation method applied in research for the analysis of the mitochondrial metabolic profile of oxidative phosphorylation (OXPHOS) in PLT obtained from small samples of human blood.

We studied PLT obtained from 29 healthy donors by high-resolution respirometry for comparison of PA and DC isolates. ROUTINE respiration and electron transfer capacity of living PLT isolated by PA were significantly higher than in the DC group, whereas plasma membrane permeabilization resulted in a 57 % decrease of succinate oxidation in PA compared to DC. These differences were eliminated after washing the PA cells with phosphate buffer containing 10 mmol·L-1 EGTA, suggesting that several components, particularly Ca2+ and fuel substrates, were carried over into the respiratory assay from the serum in PA. A simple washing step was sufficient to enable functional mitochondrial analysis in subsamples obtained from PA.

The combination of the standard clinical PA isolation procedure with PLT quality control and routine mitochondrial OXPHOS diagnostics meets an acute clinical demand in biomedical research of patients suffering from thrombocytopenia and metabolic diseases.

Keywords: mitochondria mt, human platelets PLT, thrombocytes, platelet apheresis PA, oxidative phosphorylation OXPHOS, high-resolution respirometry HRR, density gradient centrifugation DC, flow cytometry Bioblast editor: Gnaiger E O2k-Network Lab: CZ Hradec Kralove Cervinkova Z, AT Innsbruck Oroboros

ORCID: ORCID.png Vernerova A, ORCID.png Garcia-Souza LF, ORCID.png Soucek O, ORCID.png Kostal M, ORCID.png Rehacek V, ORCID.png Krcmova LK, ORCID.png Gnaiger E, ORCID.png Sobotka O

References

LinkReferenceViewYear
Braganza A, Corey CG, Santanasto AJ, Distefano G, Coen PM, Glynn NW, Nouraie SM, Goodpaster BH, Newman AB, Shiva S (2019) Platelet bioenergetics correlate with muscle energetics and are altered in older adults. JCI Insight 5:128248.PMID: 31120438 Open Access2019
Brand MD, Nicholls DG (2011) Assessing mitochondrial dysfunction in cells. Biochem J 435: 297-312.http://www.biochemj.org/bj/435/0297/bj4350297.htm2011
Bynum JA, Adam Meledeo M, Getz TM, Rodriguez AC, Aden JK, Cap AP, Pidcoke HF (2016) Bioenergetic profiling of platelet mitochondria during storage: 4°C storage extends platelet mitochondrial function and viability. Transfusion 56 Suppl 1:S76-84.PMID: 270013652016
Chacko BK, Kramer PA, Ravi S, Johnson MS, Hardy RW, Ballinger SW, Darley-Usmar VM (2013) Methods for defining distinct bioenergetic profiles in platelets, lymphocytes, monocytes, and neutrophils, and the oxidative burst from human blood. Lab Invest 93:690-700.PMID: 23528848 Open Access2013
Doerrier C, Garcia-Souza LF, Krumschnabel G, Wohlfarter Y, Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and isolated mitochondria. Methods Mol Biol 1782:31-70. doi: 10.1007/978-1-4939-7831-1_3PMID: 29850993 »O2k-brief
O2k-Protocols
2018
Ferguson MA, Sutton RM, Karlsson M, Sjövall F, Becker LB, Berg RA, Margulies SS, Kilbaugh TJ (2016) Increased platelet mitochondrial respiration after cardiac arrest and resuscitation as a potential peripheral biosignature of cerebral bioenergetic dysfunction. J Bioenerg Biomembr 48:269-79.PMID: 270205682016
Fišar Z, Jirák R, Zvěřová M, Setnička V, Habartová L, Hroudová J, Vaníčková Z, Raboch J (2019) Plasma amyloid beta levels and platelet mitochondrial respiration in patients with Alzheimer's disease. Clin Biochem 72:71-80.PMID: 309544362019
Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2: 112 pp. doi:10.26124/bec:2020-0002Bioblast pdfPublished online: 2020-Dec-30

2020
Gnaiger E, Steinlechner-Maran R, Méndez G, Eberl T, Margreiter R (1995) Control of mitochondrial and cellular respiration by oxygen. J Bioenerg Biomembr 27:583-96.PMID: 8746845
Bioblast Access
1995
Gnaiger Erich et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1:44 pp. doi:10.26124/bec:2020-0001.v1BEC 2020.1 pdfPublished online: 2020-May-20

2020
Gvozdjakova A, Sumbalova Z, Kucharska J, Chladekova A, Rausova Z, Vancova O, Komlosi M, Ulicna O, Mojto V (2019) Platelet mitochondrial bioenergetic analysis in patients with nephropathies and non-communicable diseases: a new method. Bratisl Lek Listy 120:630-35.PMID: 31475544 Open Access2019
Gvozdjakova Anna, Sumbalova Zuzana, Kucharska Jarmila, Komlosi Maria, Rausova Zuzana, Vancova Olga, Szamosova Monika, Mojto Viliam(2020) Platelet mitochondrial respiration, endogenous coenzyme Q10 and oxidative stress in patients with chronic kidney disease. Diagnostics (Basel) 10:E176.PMID: 32210203 Open Access »O2k-brief2020
Hoppel F, Calabria E, Pesta DH, Kantner-Rumplmair W, Gnaiger E, Burtscher M (2021) Effects of ultramarathon running on mitochondrial function of platelets and oxidative stress parameters: a pilot study. Front Physiol 12:632664.PMID: 33679442 Open Access »O2k-brief2021
Hsu CC, Tsai HH, Fu TC, Wang JS (2019) Exercise training enhances platelet mitochondrial bioenergetics in stroke patients: a randomized controlled trial. J Clin Med 8:E2186.PMID: 31835774 Open Access2019
Kline JA, Puskarich MA, Pike JW, Zagorski J, Alves NJ (2020) Inhaled nitric oxide to control platelet hyper-reactivity in patients with acute submassive pulmonary embolism. Nitric Oxide 96:20-8.PMID: 319405022020
McCann MR, McHugh CE, Kirby M, Jennaro TS, Jones AE, Stringer KA, Puskarich MA (2020) A multivariate metabolomics method for estimating platelet mitochondrial oxygen consumption rates in patients with sepsis. Metabolites 10:E139.PMID: 32252461 Open Access2020
Perales Villarroel JP, Figueredo R, Guan Y, Tomaiuolo M, Karamercan MA, Welsh J, Selak MA, Becker LB, Sims C (2013) Increased platelet storage time is associated with mitochondrial dysfunction and impaired platelet function. J Surg Res 184:422-9.PMID: 23830370 Open Access2013
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.PMID: 22057559
Bioblast pdf
O2k-Protocols
2012
Puskarich MA, Kline JA, Watts JA, Shirey K, Hosler J, Jones AE (2016) Early alterations in platelet mitochondrial function are associated with survival and organ failure in patients with septic shock. J Crit Care 31:63-7.PMID: 265119632016
Renner K, Geiselhöringer AL, Fante M, Bruss C, Färber S, Schönhammer G, Peter K, Singer K, Andreesen R, Hoffmann P, Oefner P, Herr W, Kreutz M (2015) Metabolic plasticity of human T cells: Preserved cytokine production under glucose deprivation or mitochondrial restriction, but 2-deoxy-glucose affects effector functions. Eur J Immunol 45:2504-16.PMID: 261142492015
Schmidl C, Renner K, Peter K, Eder R, Lassmann T, Balwierz PJ, Itoh M, Nagao-Sato S, Kawaji H, Carninci P, Suzuki H, Hayashizaki Y, Andreesen R, Hume DA, Hoffmann P, Forrest AR, Kreutz MP, Edinger M, Rehli M (2014) Transcription and enhancer profiling in human monocyte subsets. Blood 123:e90-9.PMID: 246719552014
Siewiera K, Kassassir H, Talar M, Wieteska L, Watala C (2016) Higher mitochondrial potential and elevated mitochondrial respiration are associated with excessive activation of blood platelets in diabetic rats. Life Sci 148:293-304.PMID: 268729782016
Sjoevall F, Morota S, Hansson Magnus J, Friberg H, Gnaiger E, Elmer E (2010) Temporal increase of platelet mitochondrial respiration is negatively associated with clinical outcome in patients with sepsis. Crit Care 14:R214.PMID: 21106065 Open Access2010
Sjövall F, Morota S, Asander Frostner E, Hansson Magnus J, Elmer E (2014) Cytokine and nitric oxide levels in patients with sepsis - temporal evolvement and relation to platelet mitochondrial respiratory function. PLoS One 9:e97673.PMID: 24828117 Open Access2014
Stadlmann S, Renner K, Pollheimer J, Moser PL, Zeimet AG, Offner FA, Gnaiger E (2006) Preserved coupling of oxidative phosphorylation but decreased mitochondrial respiratory capacity in IL-1ß treated human peritoneal mesothelial cells. Cell Biochem Biophys 44:179-86.PMID: 16456220
Bioblast pdf
2006
Stadlmann S, Rieger G, Amberger A, Kuznetsov AV, Margreiter R, Gnaiger E (2002) H2O2-mediated oxidative stress versus cold ischemia-reperfusion: mitochondrial respiratory defects in cultured human endothelial cells. Transplantation 74:1800-3.PMID: 124999032002
Wang L, Wu Q, Fan Z, Xie R, Wang Z, Lu Y (2017) Platelet mitochondrial dysfunction and the correlation with human diseases. Biochem Soc Trans 45:1213-23.PMID: 290549252017
O2k-Protocols
O2k-Protocols: Isolation of blood cells for HRR.
Bioblast pdf  »Versions2020-10-22

Support

Template NextGen-O2k.jpg
The study was supported by Project SVV 260 548, the University Hospital in Hradec Kralove MH CZ - DRO (UHHK, 00179906), and COST Action CA15203 MitoEAGLE. The O2k respirometers were provided by Oroboros Instruments (Innsbruck, Austria). This work was partially funded by the European Union’s Horizon 2020 research and innovation programme under grant agreement No. 859770, NextGen-O2k project (EG).


Labels: MiParea: Respiration, Instruments;methods 


Organism: Human  Tissue;cell: Platelet  Preparation: Intact cells 

Regulation: Cyt c  Coupling state: LEAK, ROUTINE, ET  Pathway: ROX  HRR: Oxygraph-2k 

SUIT-003, MitoEAGLEPublication