IOC61

O2k-Workshop on High-Resolution Respirometry - Introductory and Advanced

Schröcken, Vorarlberg, Austria 26 April - 1 May, 2011.

Hotel Körbersee

For some pictures, see IOC60

More information on IOC61: http://www.oroboros.at/index.php?nextcourse

Format

• Introductory and advanced groups will be working in parallel session, to adjust the programme optimally to the specific needs of the participants in different workpackages.

• Hot Topics in Mitochondrial Physiology: 10+5 min

Abstracts

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Combined introductory and advanced O2k-Workshop with parallel sessions

A: Introductory lectures and hands-on experiments for basic HRR in small working teams:

A1. General HRR topics

• O2k-assembly,
• Polarographic oxygen sensor (OroboPOS) service,
• POS calibration,
• DatLab software: principles and operation,
• Titration-Injection-microPump - TIP2k.

A2. Protocols in HRR

• Introduction to fundamental elements of mitochondrial respiratory physiology,
• Design and examples of experimental protocols with intact cells, permeabilized cells and tissues, isolated mitochondria.

A3. Hands-on experiments with the O2k

• Measurement of instrumental background oxygen flux, manual and TIP2k-feedback control,
• Intact cells: Respirometric reference assay with yeast, phosphorylation control protocol,
• Preparation of permeabilized muscle fibres,
• HRR with permeabilized muscle fibres.

A4. Presentation and interpretation of oxygen flux

• Normalization of respiration per tissue or per mitochondrial marker,
• Coupling control ratios,
• Substrate control ratios,
• Generalized flux control ratios.

A5. Overview on O2k-MultiSensor applications - Simultaneous measurement of respiration and:

• Mitochondrial membrane potential (TPP+ electrode),
• Acidification (pH electrode),
• Nitric oxide (amperometric),
• Spectrophotometry (cytochrome spectra),
• Spectrofluorimetry (Amplex red, safranin, etc.)

B: Advanced users will join in small working teams in parallel sessions, with specific topics:

B1. O2k-MultiSensor and TIP2k

• Hands-on MultiSensor TPP⁺ applications, see TPP special interest group during an International Oxygraph Course
• Oxygen kinetics: experimental examples with TIP2k-feedback control (oxystat) and data analysis,
• ADP kinetics: experimental examples with TIP2-steady state control and data analysis,
• pH-stat feedback control,

B2. Theoretical perspectives

• Advanced experimental protocols,
• Quality control - towards quality assurance;

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Participants and areas of interest

• Ingrid Hals, Norway: To study effects of hypoxia in pancreatic beta-cells on mitochondrial functions.
• Alberto Sanz, Finland: Mechanism causing aging and the role of mitochondria in this process.
• Takashi Yokota, Denmark: The role of mitochondria in chronic heart failure, diabetes, or obesity.
• Mikhail Alexeyev, USA: Mouse modeling of mitochondrial disorders caused by mtDNA mutations.

Tutors

• Erich Gnaiger (Innsbruck Medical University, AT; OROBOROS INSTRUMENTS, AT)
• Mario Fasching (OROBOROS INSTRUMENTS, AT)
• Dominik Pesta (Innsbruck Medical University, AT)
• Zuzana Sumbalova (OROBOROS INSTRUMENTS, AT)

• Contact: Katrin Stecher, administration; Email katrin.stecher@oroboros.at

Discussion

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