publication . Article . 2016

assessing bioenergetic functions from isolated mitochondria in drosophila melanogaster

Aw, Wen C.; Bajracharya, Rijan; Towarnicki, Samuel G.; Ballard, J. William O.;
Open Access
  • Published: 01 Jun 2016 Journal: Journal of Biological Methods, volume 3, page 42 (eissn: 2326-9901, Copyright policy)
  • Publisher: Journal of Biological Methods
Abstract
Mitochondria are involved in generating more than 90 percent of cellular energy and are responsible for many cellular processes such as metabolism, cell signalling, apoptosis and ageing. Currently, there are a number of different experimental approaches employed to measure mitochondrial health and function. Here, we demonstrate a novel approach that quantifies substrate induced mitochondrial respiration from Drosophila. This protocol is optimized for mitochondria isolated from third instar larvae, and can also be used for mitochondria isolated from adult thoraces. This procedure outlines how to perform high throughput and high resolution mitochondria specific me...
Subjects
Medical Subject Headings: fungi
free text keywords: Ageing, Apoptosis, Drosophila melanogaster, biology.organism_classification, biology, Cell signaling, Mitochondrion, Respiration, Cell biology, Bioenergetics, Metabolism, Ecology, Protocol, Drosophila, mitochondrial isolation, Seahorse XF, complex I, larvae
Related Organizations
Funded by
ARC| Discovery Projects - Grant ID: DP160102575
Project
  • Funder: Australian Research Council (ARC) (ARC)
  • Project Code: DP160102575
  • Funding stream: Discovery Projects

Scheibye-Knudsen, M., Mitchell, S. J., Fang, E. F., Iyama, T., Ward, T.. A high-fat diet and NAD(+) activate Sirt1 to rescue premature aging in cockayne syndrome. Cell Metab. 2014; 20: 840-855 [OpenAIRE] [PubMed] [DOI]

Brand, M. D., Nicholls, D. G.. Assessing mitochondrial dysfunction in cells. Biochem J. 2011; 435: 297-312 [OpenAIRE] [PubMed] [DOI]

Boutagy, N. E., Rogers, G. W., Pyne, E. S., Ali, M. M., Hulver, M. W.. Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements. J Vis Exp. 2015 [OpenAIRE] [DOI]

Rogers, G. W., Brand, M. D., Petrosyan, S., Ashok, D., Elorza, A. A.. High throughput microplate respiratory measurements using minimal quantities of isolated mitochondria. PLoS One. 2011; 6: 21746-10 [OpenAIRE] [DOI]

Das, K. C., Muniyappa, H.. Age-dependent mitochondrial energy dynamics in the mice heart: role of superoxide dismutase-2. Exp Gerontol. 2013; 48: 947-959 [OpenAIRE] [PubMed] [DOI]

Kenwood, B. M., Weaver, J. L., Bajwa, A., Poon, I. K., Byrne, F. L.. Identification of a novel mitochondrial uncoupler that does not depolarize the plasma membrane. Mol Metab. 2013; 3: 114-123 [OpenAIRE] [PubMed] [DOI]

Zhang, J., Nuebel, E., Wisidagama, D. R. R., Setoguchi, K., Hong, J. S.. Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells. Nat Protoc. 2012; 7: 1068-1085 [OpenAIRE] [PubMed] [DOI]

Joseph, A., Adhihetty, P. J., Buford, T. W., Wohlgemuth, S. E., Lees, H. A.. The impact of aging on mitochondrial function and biogenesis pathways in skeletal muscle of sedentary high- and low-functioning elderly individuals. Aging Cell. 2012; 11: 801-809 [OpenAIRE] [PubMed] [DOI]

Kuznetsov, A. V., Veksler, V., Gellerich, F. N., Saks, V., Margreiter, R.. Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells. Nat Protoc. 2008; 3: 965-976 [OpenAIRE] [PubMed] [DOI]

Pesta, D., Gnaiger, E.. High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods Mol Biol. 2012; 810: 25-58 [OpenAIRE] [PubMed] [DOI]

Wu, M., Neilson, A., Swift, A. L., Moran, R., Tamagnine, J.. Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. Am J Physiol Cell Physiol. 2006; 292: 125-136 [OpenAIRE] [DOI]

Gerencser, A. A., Neilson, A., Choi, S. W., Edman, U., Yadava, N.. Quantitative microplate-based respirometry with correction for oxygen diffusion. Anal Chem. 2009; 81: 6868-6878 [OpenAIRE] [PubMed] [DOI]

Schuh, R. A., Jackson, K. C., Khairallah, R. J., Ward, C. W., Spangenburg, E. E.. Measuring mitochondrial respiration in intact single muscle fibers. Am J Physiol Regul Integr Comp Physiol. 2011; 302: 712-9 [OpenAIRE] [DOI]

Nicholls, D. G.. The physiological regulation of uncoupling proteins. Biochim Biophys Acta. 2006; 1757: 459-466 [OpenAIRE] [PubMed] [DOI]

Abstract
Mitochondria are involved in generating more than 90 percent of cellular energy and are responsible for many cellular processes such as metabolism, cell signalling, apoptosis and ageing. Currently, there are a number of different experimental approaches employed to measure mitochondrial health and function. Here, we demonstrate a novel approach that quantifies substrate induced mitochondrial respiration from Drosophila. This protocol is optimized for mitochondria isolated from third instar larvae, and can also be used for mitochondria isolated from adult thoraces. This procedure outlines how to perform high throughput and high resolution mitochondria specific me...
Subjects
Medical Subject Headings: fungi
free text keywords: Ageing, Apoptosis, Drosophila melanogaster, biology.organism_classification, biology, Cell signaling, Mitochondrion, Respiration, Cell biology, Bioenergetics, Metabolism, Ecology, Protocol, Drosophila, mitochondrial isolation, Seahorse XF, complex I, larvae
Related Organizations
Funded by
ARC| Discovery Projects - Grant ID: DP160102575
Project
  • Funder: Australian Research Council (ARC) (ARC)
  • Project Code: DP160102575
  • Funding stream: Discovery Projects

Scheibye-Knudsen, M., Mitchell, S. J., Fang, E. F., Iyama, T., Ward, T.. A high-fat diet and NAD(+) activate Sirt1 to rescue premature aging in cockayne syndrome. Cell Metab. 2014; 20: 840-855 [OpenAIRE] [PubMed] [DOI]

Brand, M. D., Nicholls, D. G.. Assessing mitochondrial dysfunction in cells. Biochem J. 2011; 435: 297-312 [OpenAIRE] [PubMed] [DOI]

Boutagy, N. E., Rogers, G. W., Pyne, E. S., Ali, M. M., Hulver, M. W.. Using Isolated Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High throughput Microplate Respiratory Measurements. J Vis Exp. 2015 [OpenAIRE] [DOI]

Rogers, G. W., Brand, M. D., Petrosyan, S., Ashok, D., Elorza, A. A.. High throughput microplate respiratory measurements using minimal quantities of isolated mitochondria. PLoS One. 2011; 6: 21746-10 [OpenAIRE] [DOI]

Das, K. C., Muniyappa, H.. Age-dependent mitochondrial energy dynamics in the mice heart: role of superoxide dismutase-2. Exp Gerontol. 2013; 48: 947-959 [OpenAIRE] [PubMed] [DOI]

Kenwood, B. M., Weaver, J. L., Bajwa, A., Poon, I. K., Byrne, F. L.. Identification of a novel mitochondrial uncoupler that does not depolarize the plasma membrane. Mol Metab. 2013; 3: 114-123 [OpenAIRE] [PubMed] [DOI]

Zhang, J., Nuebel, E., Wisidagama, D. R. R., Setoguchi, K., Hong, J. S.. Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells. Nat Protoc. 2012; 7: 1068-1085 [OpenAIRE] [PubMed] [DOI]

Joseph, A., Adhihetty, P. J., Buford, T. W., Wohlgemuth, S. E., Lees, H. A.. The impact of aging on mitochondrial function and biogenesis pathways in skeletal muscle of sedentary high- and low-functioning elderly individuals. Aging Cell. 2012; 11: 801-809 [OpenAIRE] [PubMed] [DOI]

Kuznetsov, A. V., Veksler, V., Gellerich, F. N., Saks, V., Margreiter, R.. Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells. Nat Protoc. 2008; 3: 965-976 [OpenAIRE] [PubMed] [DOI]

Pesta, D., Gnaiger, E.. High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscle. Methods Mol Biol. 2012; 810: 25-58 [OpenAIRE] [PubMed] [DOI]

Wu, M., Neilson, A., Swift, A. L., Moran, R., Tamagnine, J.. Multiparameter metabolic analysis reveals a close link between attenuated mitochondrial bioenergetic function and enhanced glycolysis dependency in human tumor cells. Am J Physiol Cell Physiol. 2006; 292: 125-136 [OpenAIRE] [DOI]

Gerencser, A. A., Neilson, A., Choi, S. W., Edman, U., Yadava, N.. Quantitative microplate-based respirometry with correction for oxygen diffusion. Anal Chem. 2009; 81: 6868-6878 [OpenAIRE] [PubMed] [DOI]

Schuh, R. A., Jackson, K. C., Khairallah, R. J., Ward, C. W., Spangenburg, E. E.. Measuring mitochondrial respiration in intact single muscle fibers. Am J Physiol Regul Integr Comp Physiol. 2011; 302: 712-9 [OpenAIRE] [DOI]

Nicholls, D. G.. The physiological regulation of uncoupling proteins. Biochim Biophys Acta. 2006; 1757: 459-466 [OpenAIRE] [PubMed] [DOI]

Powered by OpenAIRE Open Research Graph
Any information missing or wrong?Report an Issue
publication . Article . 2016

assessing bioenergetic functions from isolated mitochondria in drosophila melanogaster

Aw, Wen C.; Bajracharya, Rijan; Towarnicki, Samuel G.; Ballard, J. William O.;