Mitochondrial Physiology
The Many Faces and Functions of an Organelle
Edited by
Erich Gnaiger
MiP
MiP2005
Mitochondrial Physiology
Schröcken, Vorarlberg, Austria
1
Mitochondrial Physiology Network 10.9: 1-152 (2005)
Mitochondrial Physiology
The Many Faces and Functions of an Organelle
Edited by
Erich Gnaiger
MiP
MiP2005
Mitochondrial Physiology
Schröcken, Vorarlberg, Austria
16-20 September 2005
4th Conference on
Mitochondrial Physiology
The Mitochondrial Physiology Society
Mitochondrial Physiology – MiP2005
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MiP2005
Organization
MiP Board
Mitochondrial Physiology Society
Chair
Erich Gnaiger (Innsbruck, Austria)
Co-Chairs Guy Brown (Cambridge, United Kingdom)
Cecilia Giulivi (Davis, USA)
Eiji Takahashi (Yamagata, Japan)
Steven Hand (Baton Rouge, USA)
Local Organizers
Erich Gnaiger (Innsbruck, Austria)
Eveline Hütter (Innsbruck, Austria)
Brigitte Haffner (Innsbruck, Austria)
Advisory Board
Susanne Arnold (Berlin, Germany)
Vilma Borutaite (Cambridge, United Kingdom)
Robert Boushel (Montreal, Canada)
Susana Cadenas (Madrid, Spain)
Keith Garlid (Oregon, USA)
Charles Hoppel (Cleveland, USA - UMDF)
Josef Houstek (Prague, Czech Republic)
Pidder Jansen-Dürr (Innsbruck, Austria)
Bernhard Kadenbach (Marburg, Germany)
Andrey Kozlov (Vienna, Austria)
Thierry Letellier (Bordeaux, France)
Heimo Mairbäurl (Heidelberg, Germany)
Heinz Osiewacz (Frankfurt, Germany)
Sergio Papa (Bari, Italy)
Richard Porter (Dublin, Ireland)
Rodrigue Rossignol (Bordeaux, France)
Patrick Schrauwen (Maastricht, The Netherlands)
Vladimir Skulachev (Moscow, Russia)
Francis Sluse (Liège, Belgium)
Anibal Vercesi (Campinas, Brasil)
Lech Wojtczak (Warszawa, Poland)
Cooperation
Competence Centre Medicine Tyrol
OROBOROS INSTRUMENTS
Focus
Continuing rigorous mitochondrial bioenergetics; integrating
molecular, cellular and organismic physiology and pathology mitochondrial function in health and disease. Presentations by
established scientists and young investigators. In the face of a
tremendously increased interest in MiP2005, the max. number of
participants was increased from 100 to 160. Irrespectively, the
number of oral presentations remains limited to provide time for
discussions.
The 3rd MiP Conference was held in 2003 (Schröcken, Austria), at
which occasion the MiP Society was formed. With your on-line
registration, you may choose to become a member of the MiP
Society:
www.mitophysiology.org
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MiP
Mitochondrial
Physiology
Society
Continuing a tradition of rigorous
mitochondrial bioenergetics
Integrating molecular, cellular and
organismic physiology and pathology
Mitochondrial Physiology
www.mitophysiology.org
The Mitochondrial Physiology Socienty has been founded in 2003, to organize MiP
Conferences and MiP Workshops. These “MiP Events” focus on common and cell-specific
functions of mitochondria at molecular, organelle, cellular, tissue and organ levels. Thus
we meet the increasing demands for integrating current developments in the
methodologically and conceptually complex field of mitochondrial physiology.
Mitochondrial research has recently undergone a dramatic renaissance, due to a whole
series of discoveries revealing its central roles in cell death, disease pathology, aging,
thermogenesis, oxidative stress, cell signalling, cellular regulation and cell bioenergetics.
Functional genomics and proteomics is putting an urgent emphasis on integrative
understanding of the link between genes/molecules and cellular/organisms functions.
Because these topics require continuous technical advances there will also be an
emphasis on new methodological developments, quantitative methods of analysis in
mitochondrial research including mathematical modeling, proteomics and the use of tools
for measuring multiple parameters in an experiment. This multidisciplinary approach
requires a new forum for high-level training courses, bringing together scientists working
in divergent disciplines, to talk together about mitochondrial function and dysfunction, to
resolve differences, to develop research agendas, to spread new knowledge and
techniques, and to develop collaborations.
Until recently, mitochondrial respiratory function was mainly studied in research
laboratories specializing on the bioenergetics of isolated mitochondria and bacteria,
energy transformation in transmembrane gradients and characterization of the various
complexes of the respiratory chain. Today these methods and concepts are expanded to
applications on intact or permeablized cells and tissue preparations, and are widely used
by groups with different research and clinical orientation, particularly in molecular cell
signalling. The series of MiP Events will help to provide the expertise necessary for
successful application of the techniques and concepts on mitochondrial bioenergetics in
the wider context of molecular and cell biology, exercise physiology, diagnosis of
mitochondrial diseases, and evaluation of mitochondrial function in oxidative stress,
ischemia-reperfusion, aging and numerous areas (such as apoptosis) of key-involvement
of mitochondria.
Erich Gnaiger, Ph.D.
Chairman, MiP Society
Department of Transplant Surgery
D. Swarovski Research Laboratory
Innsbruck Medical University
Innrain 66/6
A-6020 Innsbruck, Austria
Tel: +43 512 504 24623 (24626)
Fax: +43 512 504 24625
E-mail: erich.gnaiger@uibk.ac.at
URL: www.mitophyisology.org
Mitochondrial Physiology – MiP2005
Guy Brown, Ph.D.
Co-Chairman, MiP Society
Department of Biochemistry
University of Cambridge
Tennis Court Rd
Cambridge CB2 1QW, UK
T +44 1223 766055
F +44 1223 333342
E-mail: gcb@mole.bio.cam.ac.uk
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MiP2005
Chairpersons
receive a MiP2005 present if their session is successfully started
and completed within the time scheduled in the programme, and
if they guide discussions such that contributions are made to
clarify what MiP is.
10+5 min Presentations: Speakers receive a MiP2005 present if their presentation
proves that they selected their 10 (max. 12) best slides, if the
PowerPoint slides are readable from the far back, and if
transmission of the key message for MiP is completed within 600 s
(use of SI units is encouraged). +5 min will be reserved for
discussions. This should particularly provide poster presenters an
additional opportunity to point out connections to their related
messages.
Posters
receive a MiP2005 present if the poster reveals focus on the
essential detail and visible perspective on MiP.
YIP - Young Investigator Presentations: Students are invited to participate in the
competition for YIP awards. (1) Selection of a YIP for a 10+5 min
oral presentation is a MiP award, based on submitted abstracts.
(2) Six YIP posters are selected during the meeting for 5-min
presentations in Session 12 (5+5 min), and these young
investigators receive MiP awards.
Instrument Exhibition: Specific instrument exhibitions are organized in the lunch
breaks and evenings, in connection with poster presentations.
Schröcken,
Vorarlberg,
Austria (1269 m above sea
level).
The
lecture
hall
(Gemeindesaal) is besides
(behind) the church. Hotel
Mohnenfluh is in the front
(left), and Hotel Tannberg is
on the right.
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Contents
Mitochondrial Physiology Board .......................................................... 2
Mitochondrial Physiology and MiP2005 ................................................ 3
United Mitochondrial Disease Foundation ............................................. 7
Opening:
Session 1:
Session 2:
Session 3:
Session 4:
Session 5:
Mitochondrial Physiology: The Many Faces
and Functions of an Organelle ................................................ 9
Integrated Cardiovascular and Mitochondrial Physiology I. Skeletal Muscle, Exercise, and Endurance ........................... 13
Integrated Cardiovascular and Mitochondrial Physiology II. Oxygen Delivery, Hypoxia, and Altitude ............................. 23
Signalling To and From the Mitochondria I. Nitric Oxide and Calcium .................................................. 33
Signalling To and From the Mitochondria – II. .................................... 45
Session 6:
Organization of the Mitochondrial Respiratory Chain
and Control of Oxidative Phosphorylation ............................... 55
Respiration, Coupling, Permeability Transition, and UCPs ..................... 69
Session 7:
Oxidative Stress and Mitochondrial Function ...................................... 83
Session 8:
Mitochondria in Ischemia-Reperfusion and Cardiomyopathy ................. 95
Session 9:
Session 11:
Mitochondria and Degenerative Diseases –
I. Animal Models ...............................................................103
Mitochondria and Degenerative Diseases –
II. Patients and Human Cell Lines ........................................113
Mitochondria and Aging .................................................................121
Session 12:
Mitochondrial Physiology – an Integration .........................................131
Session 10:
Sponsors ....................................................................................133
Author Index ...............................................................................139
Addresses of Contributors and Participants ......................................144
Cover pictures
Front
Braunarl (2,648 m above sea level), the highest peak in
the area of Schröcken, Tannberg.
Back
Impressions from MiP2003, Schröcken, Vorarlberg:
Lecture hall, and walk to the Alpmuseum uf m Tannberg
with cheese and wine reception. Pictures from MiP2003
are reproduced on pages 6, 8, 68, 102, 110 and 143.
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Welcome to MiP2003 - The music group of
Schröcken provided a warm welcome to MiP2003 in
the evening after arrival. The greetings of the
conductor Heinz
Feuerstein are
translated by
Erich Gnaiger (left).
MiP2003 Session in the lecture hall - Opening lecture by Graham Kemp, chaired by Guy Brown (top right).
Kathrin Renner assists in the presentations (foreground).
Left table 1: Zdenek Drahota (edge); left side: Josef Houstek and Miriam Palacios; right side: Veronica Hollis,
Marisol Quintero, Jacques Leibowitch, Andrea Dlasková, Petr Jezek.
Left table 2: front: Pavel Jesina, Petr Pecina, Alena Vojtisková, Olga Matejková, Ondrej Kuda, Marek Böhm;
back side: Sebastian Vogt, Susanne Arnold, Dieter Brdiczka (behind), Maik Hüttemann, Bernhard
Kadenbach, Cecilia Giulivi (standing), Katrin Staniek, Hans Nohl.
Left table 3: Adrian Lambert, Peter Schönfeld, Stefanie Jarolim (behind), Julian Pakay, Franz Hartner, Julius
Liobikas (behind), Marina Jendrach, Pavel Golik, Oliver Speer (behind), Fréderic Bouillaud
(standing).
Right table 1: front: Guy Brown; left: Shey-Shing Sheu; back: Steve Hand, Michael Verkhovsky, Jordi Bermudez
(behind), Hans van Beek, Robert Boushel (edge).
Right table 2: left: Barbara Santoro; back: Daniela Curti, Zemfira Gizatullina, Irina Shabalina, Vilma Borutaite,
Jerzy Duszynski, Marzia Arese (behind).
Right table 3: left: Domenico Boffoli; back: Brigitte Haffner, Ove Eriksson (behind), Eveline Hütter, Uwe
Schlattner (behind), Fernanda Ferreira, Marko Vendelin (behind), Nazzareno Capitanio, Claudia
Piccoli (front), Eiji Takahashi (behind).
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September 9, 2005
Dear Colleagues:
On behalf of the Board of Directors and Staff of the United Mitochondrial Disease
Foundation (UMDF), we are pleased to offer these words of congratulations and support.
At UMDF, our mission is our driving force:
To promote research and education for the diagnosis, treatment,
and cure of mitochondrial disorders and to provide support for
affected individuals and families.
It is for this reason that we are proud to provide support for this important scientific
meeting which has been organized by the Mitochondrial Physiology Society under the
Chairmanship of Prof. Erich Gnaiger. We understand that good science knows no borders
and we are committed to a world united in the struggle to unravel the medical mystery of
mitochondrial disorders.
Thank you for your talent, energy, and fervent desire to help find a cure. We truly
believe that we are on the verge of medical breakthroughs that will change our
understanding of disease in the 21st century. This is a task of no small proportion; and
we must all pull together if we are to make a difference.
Again, please accept our profound thanks for your good work. Enjoy the conference, this
wonderful setting, and please remember the thousands upon thousands of children and
adults who are counting on us for a better future or, sadly, any future at all.
Yours toward a cure,
Charles A. Mohan, Jr.
Chair, Board of Directors
With all best wishes,
J. Thomas Viall
CEO/Executive Director
Mission Statement
To promote research and education for the diagnosis, treatment, and cure of
Mitochondrial Disorders and to provide support to affected individuals and families
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Discussions MiP2003 – during the sessions
(Josef Houstek and Steve Hand, top left);
and during breaks
(Erich Gnaiger, Guy
Brown and Chris
Cooper, top right).
In the sessions –
Jordi Bermudez (right),
Domenico Boffoli,
Shey-Shing Sheu,
Nazzareno Capitanio
(far right).
In the sessions – Shey-Shing
Sheu,
Veronica
Hollis,
Dieter
Brdiczka, Marek Böhm (top);
Marisol Quintero, Hans van Beek,
Sergio Papa, Petr Jezek (top right).
The MiP Society Erich
Gnaiger
(chairman,
top)
applauds at the
election of Guy
Brown, Cecilia Giulivi (left), and Eiji
Takahashi (right)
to the Committee
of the MiP Society
(MiP2003).
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Mitochondrial Physiology: The Many Faces and
Functions of an Organelle
0-01.
The many faces of mitochondiral physiology - Welcome to
MiP2005.
Erich Gnaiger1,2
1
Swarovski Research Lab., Dept. Transplant Surgery, Innsbruck Medical
2
Univ.;
OROBOROS
INSTRUMENTS,
Innsbruck,
Austria.
–
erich.gnaiger@uibk.ac.at
Intensive high-quality research is required today in mitochondrial physiology (MiP), to
bridge the gaps between molecular, biochemical, cellular, organ-level and organismic
concepts on respiration, integrating theoretical, experimental and clinical approaches. A
mission of mitochondrial physiology is to fundamentally improve our understanding of the
integrative role of bioenergetic principles and mitochondrial function in the entire context
of the intact cell. Which cell, which mitochondrion? Mitochondria have many faces and
diverse functions. It is the comparative study of mitochondria from (and in) different cell
types and tissues, from various organisms and contrasting genetic background, that
leads to validated unifying concepts in MiP. Complex feedback loops operate between the
systems of cell signaling and energy metabolism, and between the anaerobic and aerobic
components of regulation and control of metabolic flux. MiP is thus an intellecutal
challenge deeply rooted in systems biology.
An important task of MiP is the improvement of concepts and eventually
standardization of quantitative methods for the diagnosis of mitochondrial function
(health, nutrition, quality of life, aging) and mitochondrial defects (genetic and aquired
degenerative diseases, metabolic and oxidative stress, e.g. in cancer and ischemiareperfusion), to ultimately contribute to novel therapies of mitochondrial disorders. This
global perspective requires global cooperation. In the name of the Mitochondrial
Physiology Society, I want to thank the United Mitochondrial Disease Foundation (UMDF)
for their generous financial support of MiP2005. In cooperation with all MiP members,
contributors and particularly those who received financial support for their contribution to
MiP, we will make all possible efforts to turn this great support into a growing seed
spreading our contributions world-wide. “Think Mitochondria” (UMDF) and “Integrating
molecular, cellular and organismic physiology and pathology” (MiP) are common
denominators of our common interests. Whereas MiP has its main roots in fundamental
mitochondrial bioenergetics and was established to integrate the rich scientific tradition
of bioenergetics with the new challenges of mitochondrial physiology, UMDF takes the
integrative concept of “Think Mitochondria” several steps further to the clinician, to the
family, to the individual patient. UMDF provides exquisitly the “nuts and bolts” for
research in mitochondrial medicine (C.A. Mohan Jr., Think Mitochondria, Vol. 1) - MiP can
contribute by its world-wide network of scientific excellence. Without any doubt,
therefore, the expanding fields of mitochondrial physiology and mitochondrial medicine –
MiP and UMDF – can benefit mutually from an explicit cooperation.
The idea for the first MiP workshop originated during an European training course on
bioenergetics in Grenoble, France in summer 1999, and MiP1999 was held in Schröcken,
Vorarlberg, Austria in Sept. 1999, with 14 presentations and about 30 participants.
Another small MiP workshop was organized as a satellite meeting to the European
Bioenergetics Conference (EBEC) and BioThermoKinetics Meeting (BTK) in Arcachon,
France, in Sept. 2002. At that time it was considered to organize a slightly larger
conference on MiP. The Mitochondrial Phyisology Society was founded in 2003 as a
spontaneous result of the successful 3rd MiP Conference held on 12-16 September 2003
in Schröcken, with 56 submitted abstracts and 74 participants from 19 countries.
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Although MiP2005 had to be organized originally again as a zero budget meeting, the
enthusiastic interest in the 4th Conference on Mitochondrial Physiology is reflected in the
fact that participation more than doubled, with 133 abstracts and 164 participants from
30 countries – if last-minute bureaucratic problems do not hinder delegates from China,
Ukraine or Nigeria to obtain their visa for Austria. It is an explicit strategy of MiP to
encourage active participation of young investigators in our meetings. As a result, 30
Young Investigator Presentations are student contributions to MiP2005. YIPs are
supported in terms of reduced student registration fees, which was made possible in part
by the financial support of UMDF.
While the little village of Schröcken (1269 m above sea level), with 233 inhabitants
and 800 touris beds is well known for winter sports in the Arlberg region, it is quite
remote from international airports, may be inaccessible under extreme weather
conditions due to avelanches, land slides and flooded roads (such as in August 2005),
and might be difficult to reach during occasional snowfalls which may occur even in
September. Some of the participants of MiP2005 know Schröcken not only from the
previous MiP meeting [1], but from earlier conferences on Physiological and Biochemical
Microcalorimetry [2] and BioThermoKinetics [3]. Originally we did not plan to increase
the size of the present meeting significantly beyond that of MiP2003, to maintain a
discussion-oriented and personal atmosphere for the informal exchange of ideas in a
secluded alpine environment. For the increased number of presentations, it was
necessary to design a strict time schedule for 58 oral presentations and to include poster
presentations, thus avoiding parallel sessions. The time frame of the programme offers
the flexibility to organize lunch events spontaneously, including some poster flashes,
possibly a controversy discussion, round tables on specific topics, or provide simply time
for informal person-to-person interactions. In the rain: fully emphasize the
communication around the posters in the hall; in the sunshine: walks and talks - take
your poster into nature.
It is a special pleasure to welcome the many “old” and new faces to MiP2005 in
Schröcken.
1. Gnaiger E, Renner K, eds (2003) 3rd Conference on Mitochondrial Physiology. Schröcken, Austria, Sept
2003. Program and Abstracts. Mitochondr. Physiol. Network 8.10: 1-38. www.mitophysiology.org
2. Gnaiger E, ed (1988) Physiological and Biochemical Calorimetry. 6th ISMAB Conference. Veröffentl. Univ.
Innsbruck 168: 59 pp.
3. Gnaiger E, Gellerich FN, Wyss M, eds (1994) What is Controlling Life? 50 Years after Erwin Schrödinger's
‘What is Life?’ Modern Trends in BioThermoKinetics 3. Innsbruck Univ. Press 205, Innsbruck: 336 pp.
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0-02.
The future of mitochondrial physiology.
Guy C Brown
Dept. Biochem., Univ. Cambridge, Tennis Court Road, Cambridge CB2
1QW, UK. - gcb@mole.bio.cam.ac.uk
It is not entirely clear what mitochondrial physiology is, but it may
be the study of what mitochondria do.
A Brief History of Mitochondrial Physiology:
1774:
Joseph Priestly & Antoine Lavoisier, discover oxygen and respiration.
1857:
Rudolph Kölliker, pioneer of light microscope, finds mitochondria in muscle.
1890:
Richard Altmann, develops mitochondrial stain, postulates genetic
autonomy.
1898:
Carl Benda develops crystal violet as mitochondria-specific stain. Coins
name.
1912:
Otto Warburg, identifies respiratory enzyme, crude mitochondrial isolation.
1923-1933: David Keilin, identifies cytochromes and redox chain.
1940-1943: Albert Claude, isolates intact liver mitochondria and microsomes.
1948-1951: Albert Lehninger, locates β-oxidation, TCA and oxphos in mitochondria.
1952:
George Palade, uses EM to define inner, outer membranes and cristae.
1950’s:
Britton Chance, control of mitochondrial respiration.
1961:
Peter Mitchell, chemiosmotic theory.
1972:
Denham Harman, mitochondrial free radical theory of aging.
1974:
David Nicholls, mitochondrial proton leak regulates BAT heat production.
1981:
Lynn Margulis, endosymbiotic theory of mitochondrial origins.
1981:
Fred Sanger, mtDNA sequence
1986:
Kay Tanaka et al, first mtDNA diseases reported.
1996:
Xiaodong Wang, reports cytochrome c release central to apoptosis.
I have no idea what the future of mitochondrial physiology is, but I am sure it will be
as surprising and unpredictable as it’s past. I will propose some wacky ideas about
mitochondria, not because I necessarily think they are true, but because I don’t know
that they are false.
Mitochondrial viruses: can viruses hang out in mitochondria?
Mitochondrial autonomy: can mitochondria exit and enter cells?
Mitochondrial identity crisis: can mitochondria fuse with other cell membranes?
Mitochondrial differentiation: cells differentiate and change state, do mitochondria?
Mitochondrial vibrations: is energy moved around by vibrational energy transfer?
Mitochondrial lightning: is the electric field large enough to cause ionization?
0-03.
Role of intramitochondrial ROS in long-distance transmission of the
apoptotic signal.
Vladimir P Skulachev
Dept. Bioenergetics, Belozersky Inst. Physico-Chemical Biology, Moscow State Univ.,
Moscow 119992, Russia. - skulach@belozersky.msu.ru
A simple method to study interaction of two cell cultures has been developed when
two coverslips were placed side by side in a Petri dish, one coverslip with apoptogentreated cells (the inducer coverslip), and another with non-treated cell (the recipient
coverslip). TNF, staurosporine and H2O2 treatment of cells on the inducer coverslip is
shown to initiate apoptosis on the recipient coverslip. Such an effect is increased by a
catalase inhibitor aminotriazole and is arrested by addition of catalase or by pretreatment of the cells with nanomolar concentrations of mitochondria-targeted cationic
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antioxidant MitoQ, which specifically arrests the H2O2-induced apoptosis. The MitoQ
action is abolished by an uncoupler preventing accumulation of MitoQ in mitochondria. It
is concluded that ROS produced by mitochondria in the apoptotic cells initiate the H2O2
release from these cells. The H2O2 released is employed as a long-distance cell suicide
signal messenger. In processing of such a signal by the recipient cells, mitochondrial ROS
production is also involved. It is suggested that the described phenomenon may be
involved in expansion of the apoptotic region around the damaged part of the tissue at
heart attack and stroke as well as in organoptosis, i.e. disappearance of organs during
ontogenesis.
1. Pletjushkina OY, Fetisova EK, Lyamzaev KG, Ivanova OY, Domnina LV, Vyssokikh MY, Pustovidko AV,
Vasiliev JM, Murphy MP, Chernyak BV, Skulachev VP (2005) Cell Death Differ. (in press).
2. Pletjushkina OY et al. (2005) Biochemistry (Moscow) (submitted).
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