MiPNet17.17 Amplex-Mouse-brain: Difference between revisions
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|year=2013 | |year=2013 | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
|abstract= | |abstract='''O2k-Protocol for HRR and H<sub>2</sub>O<sub>2</sub> production in mouse brain mitochondria''' | ||
'''O2k-Protocol for HRR and H<sub>2</sub>O<sub>2</sub> production in mouse brain mitochondria''' | |||
High-resolution respirometry [1] is extended by MultiSensor techniques allowing the simultaneous measurement of oxygen consumption and one additional parameter (mitochondrial membrane potential, pH, Ca<sup>2+</sup>, or NO concentration [2-5]). Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H<sub>2</sub>O<sub>2</sub> production, mitochondrial membrane potential, intracellular pH, Ca<sup>2+</sup>, Mg<sup>2+</sup>, and NADH levels. | High-resolution respirometry [1] is extended by MultiSensor techniques allowing the simultaneous measurement of oxygen consumption and one additional parameter (mitochondrial membrane potential, pH, Ca<sup>2+</sup>, or NO concentration [2-5]). Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H<sub>2</sub>O<sub>2</sub> production, mitochondrial membrane potential, intracellular pH, Ca<sup>2+</sup>, Mg<sup>2+</sup>, and NADH levels. | ||
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:>> O2k-Protocols:[http://www.oroboros.at/?o2k-protocols Overall contents] | :>> O2k-Protocols:[http://www.oroboros.at/?o2k-protocols Overall contents] | ||
:>> Product: [http://www.oroboros.at/?oxygraph OROBOROS Oxygraph-2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]] | :>> Product: [http://www.oroboros.at/?oxygraph OROBOROS Oxygraph-2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]] | ||
|mipnetlab=AT_Innsbruck_OROBOROS, AT Innsbruck Gnaiger E, AT Innsbruck MitoCom | |mipnetlab=AT_Innsbruck_OROBOROS, AT Innsbruck Gnaiger E, AT Innsbruck MitoCom | ||
|articletype=MiPNet-online Publication | |articletype=MiPNet-online Publication | ||
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|articletype=MiPNet-online Publication | |articletype=MiPNet-online Publication | ||
}} | }} | ||
== Further Information == | == Further Information == | ||
:>> [http://www.oroboros.at/?h2o2_brain-mitochondria O2k-Protocols Fluorescence] | :>> [http://www.oroboros.at/?h2o2_brain-mitochondria O2k-Protocols Fluorescence] | ||
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Aguirre E, RodrΓguez-JuΓ‘rez F, Bellelli A, Gnaiger E, Cadenas S (2010) Kinetic model of the inhibition of respiration by endogenous nitric oxide in intact cells. Biochim Biophys Acta 1797: 557-565.]] | Aguirre E, RodrΓguez-JuΓ‘rez F, Bellelli A, Gnaiger E, Cadenas S (2010) Kinetic model of the inhibition of respiration by endogenous nitric oxide in intact cells. Biochim Biophys Acta 1797: 557-565.]] | ||
# [http://www.oroboros.at/index.php?mipnetanalyzer MiPNetAnalyzer] | # [http://www.oroboros.at/index.php?mipnetanalyzer MiPNetAnalyzer] | ||
# Hickey AJ, Renshaw GM, Speers-Roesch B, Richards JG, Wang Y, Farrell AP, Brauner CJ (2012) A radical approach to beating hypoxia: depressed free radical release from heart fibres of the hypoxia-tolerant epaulette shark (Hemiscyllum ocellatum). J Comp Physiol B 182: 91-100. | # [[Hickey_2012_J_Comp_Physiol_B|Hickey AJ, Renshaw GM, Speers-Roesch B, Richards JG, Wang Y, Farrell AP, Brauner CJ (2012) A radical approach to beating hypoxia: depressed free radical release from heart fibres of the hypoxia-tolerant epaulette shark (Hemiscyllum ocellatum). J Comp Physiol B 182: 91-100.]] | ||
# Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2013) Combined high-resolution respirometry and fluorometry. Validation of safranin for determination of mitochondrial membrane potential. Methods Enzymol (in press). | # Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2013) Combined high-resolution respirometry and fluorometry. Validation of safranin for determination of mitochondrial membrane potential. Methods Enzymol (in press). | ||
# [[Gnaiger 2012 MitoPathways|Gnaiger E (2012) Mitochondrial Pathways and Respiratory Control. An Introduction to OXPHOS Analysis. 3rd ed. Mitochondr Physiol Network 17.18. OROBOROS MiPNet Publications, Innsbruck: 64 pp.]] | # [[Gnaiger 2012 MitoPathways|Gnaiger E (2012) Mitochondrial Pathways and Respiratory Control. An Introduction to OXPHOS Analysis. 3rd ed. Mitochondr Physiol Network 17.18. OROBOROS MiPNet Publications, Innsbruck: 64 pp.]] |
Revision as of 14:58, 20 November 2013
[[Has title::Fasching M, Sumbalova Z, Gnaiger E (2013) O2k-Fluorometry: HRR and H2O2 production in mouse brain mitochondria. Mitochondr Physiol Network 17.17.]] |
Β» [[Has info::Open Access-MiPNet17.17.pdf ;Versions
]]
Was written by::OROBOROS (Was published in year::2013) Was published in journal::Mitochondr Physiol Network
Abstract: [[has abstract::O2k-Protocol for HRR and H2O2 production in mouse brain mitochondria
High-resolution respirometry [1] is extended by MultiSensor techniques allowing the simultaneous measurement of oxygen consumption and one additional parameter (mitochondrial membrane potential, pH, Ca2+, or NO concentration [2-5]). Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H2O2 production, mitochondrial membrane potential, intracellular pH, Ca2+, Mg2+, and NADH levels. Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber:
- Quality control of preparations by respirometric performance.
- Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP+ or fluorophores.
- Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber.
- Additional information is acquired for a limited amount of sample.
- Oxygen dependence of parameters can be studied.
Here we introduce the O2k-Fluorescence LED2-Module, describe one selected stage in its development, and show its application for the simultaneous measurement of respiration and H2O2 production of mouse brain mitochondria.
- >> O2k-Protocols:Overall contents
- >> Product: OROBOROS Oxygraph-2k, O2k-Catalogue]]
β’ O2k-Network Lab: Was published by MiPNetLab::AT_Innsbruck_OROBOROS, Was published by MiPNetLab::AT Innsbruck Gnaiger E, Was published by MiPNetLab::AT Innsbruck MitoCom
Labels: MiParea: MiP area::Respiration, MiP area::Instruments;methods
Organism: Organism::Mouse
Tissue;cell: tissue and cell::Nervous system
Preparation: Preparation::Isolated Mitochondria
Regulation: Topic::Uncoupler Coupling state: Coupling states::LEAK, Coupling states::OXPHOS, Coupling states::ETS
HRR: Instrument and method::Oxygraph-2k, Instrument and method::Fluorometry, Instrument and method::Protocol
Further Information
References
- Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopisies of human muscle. Methods Mol Biol 810: 25-58.
- Aguirre E, RodrΓguez-JuΓ‘rez F, Bellelli A, Gnaiger E, Cadenas S (2010) Kinetic model of the inhibition of respiration by endogenous nitric oxide in intact cells. Biochim Biophys Acta 1797: 557-565.
- MiPNetAnalyzer
- Hickey AJ, Renshaw GM, Speers-Roesch B, Richards JG, Wang Y, Farrell AP, Brauner CJ (2012) A radical approach to beating hypoxia: depressed free radical release from heart fibres of the hypoxia-tolerant epaulette shark (Hemiscyllum ocellatum). J Comp Physiol B 182: 91-100.
- Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2013) Combined high-resolution respirometry and fluorometry. Validation of safranin for determination of mitochondrial membrane potential. Methods Enzymol (in press).
- Gnaiger E (2012) Mitochondrial Pathways and Respiratory Control. An Introduction to OXPHOS Analysis. 3rd ed. Mitochondr Physiol Network 17.18. OROBOROS MiPNet Publications, Innsbruck: 64 pp.