IOC70 O2k-Workshop Barcelona

 

Oxygraph-2k: High-Resolution Respirometry and O2k-Fluorometry

 

Two-day O2k-Workshop presented by Erich Gnaiger

Date: 2012 May 29 - 30

Location: Esther Kopolovitz Center (CEK), IDIBAPS-Hospital Clinic de Barcelona, c/Rossello 153, Meeting rooms 1 and 2, 08036 Barcelona

        May 29: General introduction to the Oxygraph-2k. 10:00 - 13:00 h,   15:00 - 19:00 h

        May 30: The new O2k-Fluorescence LED2 module and Hot Topics in Mitochondrial Physiology. 10:00 - 13:00 h, 15:00 - 19:00 h

 

        Limited number of Participants; Registration required. All short presentations (10+5 min) are scheduled on May 30, in the late morning and late afternoon sessions.

 

Registration (informal): Dr. Pablo Garcia-Roves, Esther Kopolovitz Center (CEK), Barcelona  -  OROBOROS MipNet Reference Laboratory

 

Contribution to K-Regio project MitoCom Tyrol; information from the previous O2k-Fluorometry Workshop.



IOC70 Programme

  

1

Tuesday, May 29

 

 

O2k-Workshop

Weblink

09:45

Arrival and registration

 

 

Principles of high-resolution respirometry - from switching on the Oxygraph-2k to the experimental result

Gnaiger 2008 POS

10:00-10:05

Pablo Garcia-Roves:
  Welcome to the Madrid O2k-Workshop.

 

10:05-11:15

Erich Gnaiger:
 
The O2k-concept on high-resolution respirometry.

Glossary: Respiratory states

 

1. Get O2k-connected with your OROBOROS-USB:
a guided tour to the O2k.

get O2k-Connected

11:15

Coffee / Tea - Get-together - introduction of the participants and their research interests

POS Service

11:45-13:00

2. Oxygraph-2k: O2k-Setup and OroboPOS service.

O2k-Start

 

3. OroboPOS -  calibration, stability testing, and evaluation of sensitivity to measure oxygen flux.

O2k-Calibration

 

4. O2k-background test and analysis of oxygen flux.

O2k-Background

13:30

Lunch

 

15:00-16:30

Erich Gnaiger:
  Experimental design - Substrate and coupling control
  of mitochondrial respiration: SUIT protocols.

Gnaiger 2009 Int J Biochem Cell Biol

16:30

Coffee / Tea

CI+II

17:00-19:00

Pablo Garcia-Roves and Erich Gnaiger:
  O2k-Demoexperiment

Pesta 2012 Methods Mol Biol

 

Further information from the O2k-Manual and O2k-Protocols

 

DatLab Guide through the menues:
DL-Demo files and DL-Excel templates

DatLab 4 Guide

 

DatLab Analysis and Excel templates

DatLab4 Flux Analysis

 

Coupling control of mitochondrial respiration: LEAK, OXPHOS, ETS, ROX

mt-Respiration

 

Coupling Control Protocol with intact cells: ROUTINE, LEAK, ETS, ROX

Cell Respiration

2

Wednesday, May 30

 

 

O2k-Fluorometry and Mitochondrial Physiology

Weblink

10:00-11:30

Erich Gnaiger:
  The O2k-Fluorometer concept

 

 

1. The O2k-Fluorescence LED2-Module.

O2k-Fluorescence_LED2-Module

 

2. O2k-Fluo application with Amplex Ultrared: Oxygen dependence of hydrogen peroxide production.

O2k-Fluorescence

11:30

Coffee / Tea

 

 

Hot MiP-Topics (Chair: Pablo Garcia-Roves)

 

12:00-12:15

Perales Jose C:

 

 

Do we understand hepatic gluconeogenesis?

 

12:15-12:30

Hernandez Maribel:

 

 

Mitofusin 2 (Mfn2) links mitochondrial and endoplasmic reticulum function with insulin signaling and is essential for normal glucose homeostasis.

 

12:30-12:45

Claret Marc:

 

 

Deletion of mitochondrial fusion protein Mitofusin 2 in hypothalamic POMC neurons leads to obesity and impaired glucose homeostasis in mice.

 

12:45-13:00

Villena Josep A:

 

 

Transcriptional control of mitochondrial function in white adipose tissue by PGC-1 co-activators.

 

13:30

Lunch

 

15:00-16:30

Erich Gnaiger:
 
Permeabilized fibres vs muscle tisse homogenate

 

 

1. Are permeabilized muscle fibres a valid model for measurement of hydrogen peroxide production?

 

 

2. Mitochondrial respiratory function in tissue homogenate (PBI-Shredder) versus permeabilized fibres.

Shredder_vs_Fibres

 

3. Respiration with succinate - paradoxes in MiP.

IOC66.pdf

16:30

Coffee / Tea

 

17:00-18:00

Erich Gnaiger:

 

 

Fluorometric (AmR) and potentiometric (TPP) measurement of mt-membrane potential and future perspectives O2k-MultiSensor developments.

O2k-MultiSensor

 

Hot MiP-Topics (Chair: Pablo Garcia-Roves)

 

18:00-18:15

Vieira Elaine:

 

 

The effect of melatonin on mitochondrial respiration.

 

18:15-18:30

Garrabou Gloria:

 

 

Oxygen consumption in mitochondrial toxicity and disease.

 

18:30-18:45

Bosch Marta:

 

 

Mitochondrial cholesterol: A connection between caveolin, metabolism, and disease.

 

18:45-19:00

Summary discussion: O2k-Workshop Feedback

O2k-Feedback

 

 



IOC70 Participants

  

Name

Lab  

Alcarraz Gema 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

Bano Maria 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Secondary lesions due to chronic alcohol ingestion, muscle pathology group - ES

Bosch Marta 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Cell proliferation and signalling group - ES Barcelona Bosch M

Catalan Marc 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Secondary lesions due to chronic alcohol ingestion, muscle pathology group - ES

Claret Marc 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

De Nigris Valeria 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

Diaz-Ramos Angels 

Institute for Research in Biomedicine, IRB Barcelona -  ES 

Garcia-Roves Pablo M. 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES Barcelona Garcia-Roves PM

Garrabou Gloria 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Secondary lesions due to chronic alcohol ingestion, muscle pathology group - ES

Gnaiger Erich

DSL, Dept Visceral, Transpl Thoracic Surgery, Medical Univ Innsbruck - AT Innsbruck OROBOROS INSTRUMENTS

Gonzalez Franquesa Alba

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES Barcelona Garcia-Roves PM

Herms Albert 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Cell proliferation and signalling group - ES 

Hernandez-Alvarez Maribel

Institute for Research in Biomedicine, IRB Barcelona -  ES Barcelona Zorzano A

Lerin Carles 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

Llobet Sese Laura

Biochemistry and Molecular and Cellular Biology Department, Faculty of Veterinary University of Zaragoza, Laboratory Dr.Julio Montoya - ES

Martinez Laura 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

Martinez Lourdes Navarro

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES Barcelona Garcia-Roves PM 

Matias Nuria 

Institute of Biomedical Research of Barcelona - ES

Mendez Andres  

Bellvitge Biomedical Research Institute, IDIBELL, University of Barcelona - ES 

Moren Constanza 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Secondary lesions due to chronic alcohol ingestion, muscle pathology group - ES

Perales Jose C

Bellvitge Biomedical Research Institute, IDIBELL, University of Barcelona - ES Barcelona Bermudez MJ 

Ribas Vincente

Institute of Biomedical Research of Barcelona - ES

Schneeberger Marc 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

Tobias Ester

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Secondary lesions due to chronic alcohol ingestion, muscle pathology group - ES

Vieira Elaine 

August Pi i Sunyer Biomedical Research Institute, IDIBAPS Barcelona, Diabetes and obesity laboratory - ES

Villena Josep A

Vall D’Hebron Research Institute, VHIR Barcelona - ES

 

 




 

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