Applications in Cardiovascular

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App Notes & Tech Briefs

Assessing mitochondrial dysfunction in primary cardiomyocytes

Mitochondrial reserve capacity is a sensitive measure of cellular stress

This Application Note describes a method for profiling mitochondrial function in cells responding to stress. The mitochondrial profile generated in this way provides four parameters of mitochondrial function that can be measured in one experiment...
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Understanding the Relationship Between Bioenergetic Rates and ATP Turnover

Correlating OCR and ECAR to ATP Flux

Using the Seahorse XF24 analyzer under typical in vitro cell culture conditions, the rate of oxygen consumption (OCR) is indicative of mitochondrial respiration and the rate of extracellular acidification (ECAR) reflects glycolysis. These pathways generate ATP to power the cell...
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Publications

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The prolyl hydroxylase oxygen-sensing pathway is cytoprotective and allows maintenance of mitochondrial membrane potential during metabolic inhibition
Sridharan V, Guichard J, Bailey R M, Kasiganesan H, Beeson C and Wright G L
Am J Physiol Cell Physiol; 2007 Feb 1. 292 (2):C719-728.
ERRgamma directs and maintains the transition to oxidative metabolism in the postnatal heart
Alaynick W A, Kondo R P, Xie W, He W, Dufour C R, Downes M, Jonker J W, Giles W, Naviaux R K, Giguere V and Evans R M
Cell Metab; 2007 Jul 1. 6 (1):13-24.
Hypoxia-inducible factor induces local thyroid hormone inactivation during hypoxic-ischemic disease in rats
Simonides W S, Mulcahey M A, Redout E M, Muller A, Zuidwijk M J, Visser T J, Wassen F W, Crescenzi A, da-Silva W S, Harney J, Engel F B, Obregon M J, Larsen P R, Bianco A C and Huang S A
J Clin Invest; 2008 Mar 1. 118 (3):975-983.
O(2)-sensing signal cascade: clamping of O(2) respiration, reduced ATP utilization, and inducible fumarate respiration
Sridharan V, Guichard J, Li C Y, Muise-Helmericks R, Beeson C C and Wright G L
Am J Physiol Cell Physiol; 2008 Jul 1. 295 (1):C29-37.
MicroRNA-210 controls mitochondrial metabolism during hypoxia by repressing the iron-sulfur cluster assembly proteins ISCU1/2
Chan S Y, Zhang Y Y, Hemann C, Mahoney C E, Zweier J L and Loscalzo J
Cell Metab; 2009 Oct 1. 10 (4):273-284.
Importance of the bioenergetic reserve capacity in response to cardiomyocyte stress induced by 4-hydroxynonenal
Hill B G, Dranka B P, Zou L, Chatham J C and Darley-Usmar V M
Biochem J; 2009 Nov 15. 424 (1):99-107.
Regulation of vascular smooth muscle cell bioenergetic function by protein glutathiolation
Hill B G, Higdon A N, Dranka B P and Darley-Usmar V M
Biochim Biophys Acta; 2010 Feb 1. 1797 (2):285-295.
Nutrient-sensitized screening for drugs that shift energy metabolism from mitochondrial respiration to glycolysis
Gohil V M, Sheth S A, Nilsson R, Wojtovich A P, Lee J H, Perocchi F, Chen W, Clish C B, Ayata C, Brookes P S and Mootha V K
Nat Biotechnol; 2010 Mar 1. 28 (3):249-255.
Mitochondrial reserve capacity in endothelial cells: The impact of nitric oxide and reactive oxygen species
Dranka B P, Hill B G and Darley-Usmar V M
Free Radic Biol Med; 2010 Apr 1. 48 (7):905-914.
Role of cellular bioenergetics in smooth muscle cell proliferation induced by platelet-derived growth factor
Perez J, Hill B G, Benavides G A, Dranka B P and Darley-Usmar V M
Biochem J; 2010 Jun 1. 428 (2):255-267.
Loss of hypoxia-inducible factor prolyl hydroxylase activity in cardiomyocytes phenocopies ischemic cardiomyopathy
Moslehi J, Minamishima Y A, Shi J, Neuberg D, Charytan D M, Padera R F, Signoretti S, Liao R and Kaelin W G, Jr.
Circulation; 2010 Sep 7. 122 (10):1004-1016.
Responses of hypertrophied myocytes to reactive species: implications for glycolysis and electrophile metabolism
Sansbury B E, Riggs D W, Brainard R E, Salabei J K, Jones S P and Hill B G
Biochem J; 2011 Apr 15. 435 (2):519-528.
Bioenergetic function in cardiovascular cells: The importance of the reserve capacity and its biological regulation
Sansbury B E, Jones S P, Riggs D W, Darley-Usmar V M and Hill B G
Chem Biol Interact; 2011 May 30. 191 (1-3):288-295.
Mitochondria determine the differentiation potential of cardiac mesoangioblasts
San Martin N, Cervera A M, Cordova C, Covarello D, McCreath K J and Galvez B G
Stem Cells; 2011 Jul 1. 29 (7):1064-1074.
Atrial natriuretic peptide regulates lipid mobilization and oxygen consumption in human adipocytes by activating AMPK
Souza S C, Chau M D, Yang Q, Gauthier M S, Clairmont K B, Wu Z, Gromada J and Dole W P
Biochem Biophys Res Commun; 2011 Jul 8. 410 (3):398-403.
Mitochondrial Transporter ATP Binding Cassette Mitochondrial Erythroid Is a Novel Gene Required for Cardiac Recovery After Ischemia/Reperfusion
Liesa M, Luptak I, Qin F, Hyde B B, Sahin E, Siwik D A, Zhu Z, Pimentel D R, Xu X J, Ruderman N B, Huffman K D, Doctrow S R, Richey L, Colucci W S and Shirihai O S
Circulation; 2011 Aug 16. 124 (7):806-813.
Hydrogen sulfide replacement therapy protects the vascular endothelium in hyperglycemia by preserving mitochondrial function
Suzuki K, Olah G, Modis K, Coletta C, Kulp G, Gero D, Szoleczky P, Chang T, Zhou Z, Wu L, Wang R, Papapetropoulos A and Szabo C
Proc Natl Acad Sci U S A; 2011 Aug 16. 108 (33):13829-13834.
Differential regulation of metabolism by nitric oxide and S-nitrosothiols in endothelial cells
Diers A R, Broniowska K A, Darley-Usmar V M and Hogg N
Am J Physiol Heart Circ Physiol; 2011 Sep 1. 301 (3):H803-812.
Identification and characterization of a functional mitochondrial angiotensin system
Abadir P M, Foster D B, Crow M, Cooke C A, Rucker J J, Jain A, Smith B J, Burks T N, Cohn R D, Fedarko N S, Carey R M, O'Rourke B and Walston J D
Proc Natl Acad Sci U S A; 2011 Sep 6. 108 (36):14849-14854.
The Transcriptional Coactivators, PGC-1? and ?, Cooperate to Maintain Cardiac Mitochondrial Function During the Early Stages of Insulin Resistance
Mitra R, Nogee D P, Zechner J F, Yea K, Gierasch C M, Kovacs A, Medeiros D M, Kelly D P and Duncan J G
J Mol Cell Cardiol. 2011 Oct 20; [Epub ahead of print].
GLUT10 is required for the development of the cardiovascular system and the notochord and connects mitochondrial function to TGFbeta signaling
Willaert A, Khatri S, Callewaert B L, Coucke P J, Crosby S D, Lee J G, Davis E C, Shiva S, Tsang M, De Paepe A and Urban Z
Hum Mol Genet. 2011 Nov 24; [Epub ahead of print].
Susceptibility to simvastatin-induced toxicity is partly determined by mitochondrial respiration and phosphorylation state of Akt
Mullen P J, Zahno A, Lindinger P, Maseneni S, Felser A, Krahenbuhl S and Brecht K
Biochim Biophys Acta; 2011 Dec 1. 1813 (12):2079-87.
Poly(ADP-ribose) polymerase-2 depletion reduces doxorubicin-induced damage through SIRT1 induction
Szanto M, Rutkai I, Hegedus C, Czikora A, Rozsahegyi M, Kiss B, Virag L, Gergely P, Toth A and Bai P
Cardiovasc Res; 2011 Dec 1. 92 (3):430-438.
Molecular Analysis of Mitochondrial Compromise in Rodent Cardiomyocytes Exposed Long Term to Nucleoside Reverse Transcriptase Inhibitors (NRTIs)
Liu Y, Nguyen P, Baris T Z and Poirier M C
Cardiovasc Toxicol. 2011 Dec 15; [Epub ahead of print].
Therapeutic inhibition of fatty acid oxidation in right ventricular hypertrophy: exploiting Randle's cycle
Fang Y H, Piao L, Hong Z, Toth P T, Marsboom G, Bache-Wiig P, Rehman J and Archer S L
J Mol Med (Berl); 2012 Jan 1. 90 (1):31-43.

Technical Posters

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Snf1-related Kinase Enhances Cardiomyocyte Metabolism and Increases Mitochondrial Membrane Potential and Glycolysis Through Tribbles Homolog 3 Downreg
Amy K. Rines, Michael A. Burke, Hossein Ardehali
American Heart Association Scientific Sessions 2011
November 12-16, 2011
Orlando, FL
Endothelial Nitric Oxide Synthase Promotes a Shift in Adipocyte Phenotype in Diet-Induced Obesity
Brian E. Sansbury, Jason Hellmann, Yunan Tang, Kathie Nichol, Matthew Spite, Aruni Bhatnagar, and Bradford G. Hill
American Heart Association Scientific Sessions 2011
November 12-16, 2011
Orlando, FL
Manipulating Myocardial Wnt Signalling For Post-Ischemic Myocardial Protection
K Venardos; D Williams; D Horlock; J Peart; S McGee; D Kaye
American Heart Association Scientific Sessions 2011
November 12-16, 2011
Orlando, FL
Mitochondrial Oxidative Stress Corrupts Tube Formation in Human Coronary Artery Endothelial Cells by Inhibiting mTOR Signaling
Pung, PhD, Yuh Fen
Arteriosclerosis, thrombosis, and vascular biology, 2011 Scientific Sessions
April 28-30, 2011
Chicago, IL
Novel insights into the role of glycolysis in vascular smooth muscle cell phenotype switching
Joshua Salabei, Aruni Bhatnagar, and Bradford G. Hill
Experimental Biology 2011
April 9-13, 2011
Washington, DC, US
Bioenergetic Dysfunction in Endothelial Cells Precedes Cell Death Due to Hemin: Implications for Hemolytic Anemias
Ashlee N. Higdon, Gloria A. Benavides, Victor M. Darley-Usmar
Society for Free Radical Biology's 17th Annual Meeting
November 17-20, 2010
Orlando, FL
Simvastatin: The Enemy Within?
PJ Mullen, A Zahno, S Maseneni, S Krahensbuhl, P Lindinger
European Muscle Conference, XXXIX
September 14, 2010
Padua, IT
Inhibition of Apoptosis Partially Restores Mitochondrial Function
Gloria A. Benavides, Blake R. Zelickson, Sarah A. Owusu, Samantha Giordano, Lonnie Schneider, Brian Benoit, Jianhua Zhang and Victor M. Darley-Usmar
5th International Conference of Neurons and Brain Disease
June 29-1, 2010
Bristol, GB
Hypoxia/Reoxygenation Alters Mitochondrial Function in Endothelial Cells
Blake R. Zelickson, Gloria A. Benavides, Aimee Landar, Jianhua Zhang, and Victor M. Darley-Usmar
5th International Conference of Neurons and Brain Disease
June 29-1, 2010
Bristol, GB
Bioenergetics Changes Induced By Hydrogen Peroxide Exposure in Endothelial Cells
Gloria A. Benavides, Brian P. Dranka, Brian Benoit, and Victor M. Darley-Usmar
Society for Free Radical Biology's 16th Annual Meeting
November 18-22, 2009
San Francisco, CA
Direct Measurement of Bioenergetic Stress in Cardiomyocytes Exposed to Hydrogen Peroxide and Protection by O-GlcNAc
Brian P. Dranka, Bradford G. Hill, Luyun Zou, John C. Chatham, and Victor M. Darley-Usmar
Society for Free Radical Biology's 15th Annual Meeting
November 16-26, 2008
Indianapolis, IN
Direct measurement of bioenergetic stress in cardiomyocytes exposed to hydrogen peroxide and protection by O-GlcNAc
Brian P. Dranka, Bradford G. Hill, Luyun Zou, John C. Chatham, and Victor M. Darley-Usmar
Society for Free Radical Biology's 15th Annual Meeting
November 16-26, 2008
Indianapolis, IN

Webinars

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XF Glycolysis Stress Test Assesses Glycolysis and Glycolytic Capacity in Cells
Min Wu, PhD, Seahorse Bioscience
August 30, 2011
Mitochondrial Profiling in Cells Under Physiological Conditions and In Response to Stress
Victor Darley-Usmar, PhD, University of Alabama at Birmingham
August 4, 2010
Modulating Mitochondrial Metabolism as a Protective Strategy in Cardiac Ischemic Injury
Paul Brookes, PhD, University of Rochester Medical Center
May 20, 2010
Importance of the Mitochondrial Reserve Capacity in Cardiovascular Cells Exposed to Oxidative Stress.
Brad Hill, PhD, Center for Diabetes and Obesity Research, University of Louisville
January 20, 2010
The Response of the Mitochondrion to Oxidative Stress in Cardiovascular Cells
Victor Darley-Usmar, PhD, University of Alabama at Birmingham
November 11, 2008
Tutorial 2: Measuring Bioenergetics in Primary Cell Types
David Ferrick, PhD, CSO, Seahorse Bioscience
August 27, 2008
Protective Bioenergetic Responses of Cardiomyocytes to Hypoxic Stress
Gary L. Wright, PhD, University of South Carolina
May 29, 2008
Fueling Physiology: How Bioenergetics Drive Biological Function
Craig Beeson, PhD , Medical University of South Carolina
January 19, 2008

Mitochondria play a significant role in the regulation of cardiovascular cell function. A developing hypothesis among cardiovascular disease development and cardiovascular disease risk factors is increased mitochondrial damage and dysfunction.

The XF Analyzer generates data that can provide new understanding in relation to cardiovascular disease and mitochondrial dysfunction, by allowing the measurement of cellular metabolism in real-time, in a microplate. By measuring the oxygen consumption rate (OCR) - a measure of mitochondrial respiration; as well as the extracellular acidification rate (ECAR) - a measure of glycolysis, you will be provided with the most physiologically relevant in vitro measurement for your bioenergetic studies.