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1. Experimental assessment of bioenergetic differences caused by the common European mitochondrial DNA haplogroups H and T

Author

Taku Amo, Richard J. Oh, Martin D. Brand, Nagendra Yadava, and David G. Nicholls

Subjects
Nuclear gene, Bioenergetics, genetic structures, Haplogroup H, Cybrid, Oxidative phosphorylation, Mitochondrion, Biology, ρ0, mtDNA-less, Hybrid Cells, DNA, Mitochondrial, Haplogroup, Article, Oxidative Phosphorylation, White People, Oxygen Consumption, Δψ, mitochondrial membrane potential, Sepsis, Cell Line, Tumor, Genetics, Humans, Membrane Potential, Mitochondrial, Haplotype, Genetic Variation, General Medicine, social sciences, Molecular biology, TPMP, triphenylmethylphosphonium, eye diseases, humanities, FCCP, carbonyl cyanide p-trifluoromethoxyphenylhydrazone, Sperm motility, Haplotypes, geographic locations, Human mitochondrial DNA haplogroup, Coupling efficiency
Abstract

Studies of both survival after sepsis and sperm motility in human populations have shown significant associations with common European mitochondrial DNA haplogroups, and have led to proposals that mitochondria bearing haplogroup H have different bioenergetic capacities than those bearing haplogroup T. However, the validity of such associations assumes that there are no non-random influences of nuclear genes or other factors. Here, we removed the effect of any differences in nuclear genes by constructing transmitochondrial cybrids harbouring mitochondria with either haplogroup H or haplogroup T in cultured A549 human lung carcinoma cells with identical nuclear backgrounds. We compared the bioenergetic capacities and coupling efficiencies of mitochondria isolated from these cells, and of mitochondria retained within the cells, as a critical experimental test of the hypothesis that these haplogroups affect mitochondrial bioenergetics. We found that there were no functionally-important bioenergetic differences between mitochondria bearing these haplogroups, using either isolated mitochondria or mitochondria within cells.

Published
2008

2. Quantitative microplate-based respirometry with correction for oxygen diffusion

Author

Nagendra Yadava, Sung W. Choi, Richard J. Oh, David G. Nicholls, David A. Ferrick, Akos A. Gerencser, Andy Neilson, Ursula Edman, and Martin D. Brand

Subjects
Male, Spectrum analyzer, Diffusion, Analytical chemistry, chemistry.chemical_element, Mitochondria, Liver, Oxygen, Fluorescence, Article, Analytical Chemistry, law.invention, 03 medical and health sciences, Respirometry, Mice, 0302 clinical medicine, Oxygen Consumption, law, Animals, Clark electrode, Cells, Cultured, 030304 developmental biology, 0303 health sciences, Chemistry, Volume (thermodynamics), Respirometer, Flux (metabolism), 030217 neurology & neurosurgery, Algorithms, Synaptosomes
Abstract

Respirometry using modified cell culture microplates offers an increase in throughput and a decrease in biological material required for each assay. Plate based respirometers are susceptible to a range of diffusion phenomena; as O(2) is consumed by the specimen, atmospheric O(2) leaks into the measurement volume. Oxygen also dissolves in and diffuses passively through the polystyrene commonly used as a microplate material. Consequently the walls of such respirometer chambers are not just permeable to O(2) but also store substantial amounts of gas. O(2) flux between the walls and the measurement volume biases the measured oxygen consumption rate depending on the actual [O(2)] gradient. We describe a compartment model-based correction algorithm to deconvolute the biological oxygen consumption rate from the measured [O(2)]. We optimize the algorithm to work with the Seahorse XF24 extracellular flux analyzer. The correction algorithm is biologically validated using mouse cortical synaptosomes and liver mitochondria attached to XF24 V7 cell culture microplates, and by comparison to classical Clark electrode oxygraph measurements. The algorithm increases the useful range of oxygen consumption rates, the temporal resolution, and durations of measurements. The algorithm is presented in a general format and is therefore applicable to other respirometer systems.

Published
2009

3. Contributors

Author

Faustino Abascal, Judith E. Adams, Joong Mo Ahn, Jorge Albareda, Gina M. Allen, Arash Anavim, Mark W. Anderson, Suzanne E. Anderson, Francisco Aparisi, Pilar Aparisi, Derek R. Armfield, Bridget Atkins, Lisa O. Ballehr, Romulo Baltazar, Laura W. Bancroft, Michelle S. Barr, D. Barron, Andrea Baur-Melnyk, Francesca Beaman, Javier Beltran, Luis S. Beltran, Jenny T. Bencardino, Thomas H. Berquist, M. Biervliet, Donna G. Blankenbaker, Hans L. Bloem, Marcy B. Bolster, Peyman Borghei, Robert Downey Boutin, Patrick A. Brouwer, Ángel Bueno, Michele Calleja, Ana Canga, Victor N. Cassar-Pullicino, Conrado F.A. Cavalcanti, Luis Cerezal, Patrick Chastanet, Qi Chen, Yvonne Y. Cheung, Aaron Cho, Christine B. Chung, Claire Coggins, Anne Cotten, Winnie Courtens, Christian Czerny, Richard H. Daffner, A. Mark Davies, Kirkland W. Davis, José Luis del Cura, Bradley N. Delman, Michel O. De Maeseneer, Arthur M. De Schepper, Sachin Dheer, Rodrigo Dominguez, Davide Donati, A. Bassem Elaini, Georges Y. El-Khoury, Chris Fang, J. Farrant, Laura M. Fayad, Pilar Ferrer, Steven Finden, Donald J. Flemming, Jürgen Freyschmidt, Michael S. Gibson, Louis A. Gilula, Angela Gessner Gopez, Punita Gupta, Andrew Haims, John H. Harris, Curtis W. Hayes, Tamara Miner Haygood, Victoria Higueras, Siegfried Hofmann, Peter A. Hrehorovich, Ana M. Hualde, Wouter C.J. Huysse, Hakan Ilaslan, Karsten Jablonka, Bryan T. Jennings, Ann M. Johnson, Karl Johnson, Apostolos Karantanas, David Karasick, Theodoros Katsivas, Eoin C. Kavanagh, Theodore E. Keats, Shah H.M. Khan, Digna R. Kool, George Koulouris, Josef Kramer, Herman M. Kroon, Cornelis van Kuijk, Susanne W.Y. Lardenoye-Bröker, Hans Peter Ledermann, Gerwin M. Lingg, Brandon Y. Liu, Patrick T. Liu, Eva Llopis, Calvin T. Ma, Matthew A. Marcus, José Martel, Paul Marten, Manesh Mathew, Iain W. McCall, Eugene G. McNally, José M. Mellado, Theodore T. Miller, Douglas N. Mintz, Johnny U.V. Monu, Sandra L. Moore, William B. Morrison, Timothy J. Mosher, Michael E. Mulligan, Mark D. Murphey, William A. Murphy, Daniel B. Nissman, George C. Nomikos, Willem R. Obermann, Philip J. O'Connor, Amaka Offiah, Richard J. Oh, Imran M. Omar, Oleg Opsha, Simon Ostlere, Joshua M. Owen, William E. Palmer, Mario Padron, Narayan Babu Paruchuri, Mini N. Pathria, Rogerich T. Paylor, Christian W. Pfirrmann, Michael Pharoah, Marc J. Philippon, Thomas Lee Pope, Linda J. Probyn, Mahvash Rafii, James J. Rankine, Nisha Rao, Vijay M. Rao, Lisa Georgianne Rider, Catherine C. Roberts, Lee F. Rogers, Alejandro U. Rolón, Zehava Sadka Rosenberg, Lorne Rosenbloom, Daniel Rosenthal, David A. Rubin, Rodrigo A. Salgado, Barry Schenk, Anna Scheurecker, Jean Schils, Corinna Schorn, Mark E. Schweitzer, Jon K. Sekiya, Maryam Shahabpour, Steven Shankman, Roger Smith, Annemie Snoeckx, Travis G. Snyder, Peter M. Som, Raphaëlle Souillard, Alan Sprigg, Alison R.I. Spouge, Lynne S. Steinbach, Murali Sundaram, Mihra S. Taljanovic, James Teh, Jamshid Tehranzadeh, Bernhard Johannes Tins, Dechen W. Tshering, Michael J. Tuite, Hilary Umans, Bruno C. Vande Berg, Filip M. Vanhoenacker, Catharina S.P. van Rijswijk, Koenraad L. Verstraete, Iain Watt, Lawrence M. White, Helen J. Williams, Kevin R. Willits, David John Wilson, Emad Yacoub, Lawrence Yao, Joseph S. Yu, Natalie Zelenko, Michael B. Zlatkin, and Adam C. Zoga

Published
2008
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