Calcium signalling: Approaches and Findings in the Heart and Blood

246 Pages·2013·6.577 MB·English

by  Lars Kaestner (auth.)

Calcium signalling
Lars Kaestner
Calcium
signalling
Approaches and Findings in the Heart and Blood
With 96 figures
1 23
Dr. Lars Kaestner
Universität des Saarlandes
Institut für Molekulare Zellbiologie
ISBN-13 978-3-642-34616-3 ISBN 978-3-642-34617-0 (eBook)
DOI 10.1007/978-3-642-34617-0
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V

Foreword
This book is a habilitation treatise. I aimed to both fulfil the formal requirements of the ha-
bilitation treatise as defined in the habilitation regulation of the Medical Faculty at Saarland
University and provide a resource for colleagues, students and all that are interested in cal-
cium signalling and/or optical imaging methods to record it. As such, I hope this booklet is
more than a file in academic qualification, and that it might also serve as a useful tool when-
ever the universal messenger calcium crosses the track.
The biological examples are recruited from two major fields, cellular cardiology and red
cell research. I am grateful to the people who not only introduced me to these subjects, but ac-
companied my research for many years, who are Prof. Peter Lipp and Prof. Ingolf Bernhardt,
respectively. Although both fields seem to be independent, future research, systems biology
approaches and translational methods will reveal interconnections going far beyond the obvi-
ous ones described in the Introduction. Therefore, the current compilation can only be seen
as a snapshot, and I am looking forward to the future gain of knowledge.
Finally, I would like to thank all of my colleagues who accompanied my work and have
contributed to the scientific outcome so far. I am grateful to my family, who suffer from re-
strictions caused by the intensity of the scientific passion. Last, but not least, I thank Springer
for publishing this script.
Homburg/Saar, September 2012 Lars Kaestner
VII
Contributors
Parts II-IV contain contributions of the following people:
Bennekou, Poul Fleischmann, Bernd K. Jung, Achim
Department of Biology Life & Brain Center SECTOR Cert GfZmbH
University of Copenhagen University of Bonn Cologne
Denmark Germany Germany
Bernhardt, Ingolf Flockerzi, Veit Jung, Jennifer
Laboratory of Biophysics General & Clinical Pharmacology Life Science
Saarland University Saarland University University of Applied Science
Saarbrücken Homburg/Saar Zweibrücken
Germany Germany Germany
Bloch, Wilhelm Freichel, Marc Justus, Isabel
Molecular and Cellular Sports Me- Institute of Pharmacology Pharmacy Board
dicine University Heidelberg Bremen
German Sport University Cologne Germany Germany
Germany
Gassmann, Max Kahl, Valentin
Bogdanova, Anna Veterenary Physiology Ibidi GmbH
Veterenary Physiology University Zürich Munich
University Zürich Switzerland Germany
Switzerland
Hammer, Karin Kirchhefer, Uwe
Boldyrev, Alexander A. Department of Pharmacology Pharmacology & Toxikology
International Biotechnilogy Centre UC Davis Wilhelms University
Moscow State University USA Münster
Russia Germany
Hanske, Gabriela
Chien, Kenneth R. Aesculap Pharmacy Kraegeloh, Annette
Massachusetts General Hospital Dortmund Leibnitz Institute for New Materials
Boston Germany Saarbrücken
USA Germany
Held, Brigitte
Christophersen, Palle GRADE BioMed First Kraushaar, Udo
Neurosearch A/S Goethe University Natural and Medical Science In-
Copenhagen Frankfurt/Main stitute
Denmark Germany Reutlingen
Germany
Edelmann, Ludwig Jones, Larry R.
Molecular Cell Biology Krannert Institute of Cardiology Lipp, Peter
Saarland University Indiana University Molecular Cell Biology
Homburg/Saar Indianapolis Saarland University
Germany USA Homburg/Saar
Germany
VIII Contributors
Maia, Sara Homburg/Saar Viero, Cedric
Laboratory of Biophysics Germany Molecular and Experimental
Saarland University Medicine
Saarbrücken Ruppenthal, Sandra Cardiff University
Germany Molecular Cell Biology United Kingdom
Saarland University
Makhro, Asya Homburg/Saar Vogel, Johannes
Veterenary Physiology Germany Veterenary Physiology
University Zürich University Zürich
Switzerland Schmitz, Wilhelm Switzerland
Pharmacology & Toxikology
Müller, Oliver Wilhelms University Wagner, Christian
Institute for Genetics Münster Experimental Physics
University of Cologne Germany Saarland University
Germany Saarbrücken
Scholz, Anke Germany
Müller, Torsten Molecular Cell Biology
JPK Instruments Saarland University Wagner-Britz, Lisa
Berlin Homburg/Saar Laboratory of Biophysics
Germany Germany Saarland University
Saarbrücken
Neumann, Joachim Schumann, Christian Germany
Pharmacology & Toxikology Leica Microsystems CMS GmbH
Martin-Luther University Wetzlar Wang, Jue
Halle/Saale Germany Molecular Cell Biology
Germany Saarland University
Schwarz, Sarah Homburg/Saar
Bach Nguyen, Duc Ursapharm Arzneimittel GmbH Germany
Molecular Biology Saarbrücken
Hanoi University of Agriculture Germany Weiss, Erwin
Vietnam Veterinary Medicine
Steffen, Patrick University of Cambridge
Oberhofer, Martin Experimental Physics United Kingdom
Molecular Cell Biology Saarland University
Saarland University Saarbrücken Weissgerber, Petra
Homburg/Saar Germany Experimental Pharmacology
Germany Saarland University
Tabellion, Wiebke Homburg/Saar
Oleinikow, Katharina Roche Diagnostics GmbH Germany
Life Science Mannheim
University of Applied Science Germany Zantl, Roman
Zweibrücken Ibidi GmbH
Germany Tian, Qinghai Munich
Molecular Cell Biology Germany
Pahlavan, Sara Saarland University
Molecular Cell Biology Homburg/Saar
Saarland University Germany
IX
Content
I Summary II Calcium Signalling
Methodology
1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . 2
12 Non-linear and ultra high-speed
2 Direct evidence - the digital approach . . 3 imaging for explorations of the murine
and human heart . . . . . . . . . . . . . . . . . . . . . 36
3 Fluorescence-based visualisation . . . . . . . 4 12.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
3.1 Calcium sensors . . . . . . . . . . . . . . . . . . . . . . . . . . . 4 12.2 Methodology . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
3.2 Spatio-temporal considerations . . . . . . . . . . . . 5 12.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
3.3 Confocal scanning techniques . . . . . . . . . . . . . 6 12.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42
3.4 Further optical sectioning techniques . . . . . . 8 12.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
3.5 Super-resolution imaging techniques . . . . 12 12.6 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
4 Aequorin-based measurements . . . . . . . 14 13 A system for optical high
resolution screening of electrical
5 Measurement of calcium transport excitable cells . . . . . . . . . . . . . . . . . . . . . . . . 45
across membranes . . . . . . . . . . . . . . . . . . . . 15 13.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
5.1 Flux measurements . . . . . . . . . . . . . . . . . . . . . . 15 13.2 Materials and methods . . . . . . . . . . . . . . . . . . 46
5.2 The patch-clamp technique . . . . . . . . . . . . . . 15 13.3 Results and discussion . . . . . . . . . . . . . . . . . . . 48
13.4 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
6 Molecular biology based approaches . . 17 13.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57
6.1 Determination of molecular identities . . . . 17
6.2 Antibody-based techniques . . . . . . . . . . . . . 17 14 Concepts for optical high content
6.3 Genetic manipulation . . . . . . . . . . . . . . . . . . . 17 screens of excitable primary isolated
cells . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
7 Manipulation of calcium . . . . . . . . . . . . . . 19 14.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
7.1 Pharmacological manipulation . . . . . . . . . . . 19 14.2 Methodology . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
7.2 Optical manipulation . . . . . . . . . . . . . . . . . . . . 19 14.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
7.3 Combined approaches . . . . . . . . . . . . . . . . . . 19 14.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
14.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
8 Calcium-induced function . . . . . . . . . . . . 21 14.6 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
8.1 The concept of calcium-induced
function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 15 A primary culture system for
8.2 Cardiac contraction . . . . . . . . . . . . . . . . . . . . . . 21 sustained expression of a calcium
8.3 Red blood cell aggregation . . . . . . . . . . . . . . 21 sensor in preserved adult rat
ventricular myocytes . . . . . . . . . . . . . . . . . 67
9 Calcium signalling in cardiac 15.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
myocytes . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24 15.2 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
15.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
10 Calcium signalling in red blood cells . . . 26 15.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 80
15.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 82
11 Perspective . . . . . . . . . . . . . . . . . . . . . . . . . . 29
11.1 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
X Content
16 Calcium imaging of individual 20 Functional and morphological
erythrocytes: Problems and preservation of adult ventricular
approaches . . . . . . . . . . . . . . . . . . . . . . . . . . 83 myocytes in culture by sub-micromolar
16.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83 cytochalasin D supplement . . . . . . . . . 135
16.2 Material and methods . . . . . . . . . . . . . . . . . . . 84 20.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
16.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84 20.2 Material and methods . . . . . . . . . . . . . . . . . . 136
16.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 89 20.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137
16.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90 20.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 146
20.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 148
III Calcium Signalling in
Cardiac Myocytes IV Calcium Signalling in Red
Blood Cells
17 Reduced Cardiac L-Type Ca2+ Current
in Ca ß -/- Embryos Impairs Cardiac 21 The non-selective voltage-activated
v 2
Development and Contraction With cation channel in the human red
Secondary Defects in Vascular blood cell membrane: reconciliation
Maturation . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 between two conflicting reports
17.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 94 and further characterisation . . . . . . . . 152
17.2 Materials and Methods . . . . . . . . . . . . . . . . . . 95 21.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
17.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96 21.2 Experimental . . . . . . . . . . . . . . . . . . . . . . . . . . . 153
17.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 103 21.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 154
17.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105 21.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 158
21.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
18 Overexpression of junctin causes 21.6 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
adaptive changes in cardiac 21.7 Appendix: A Calculation of the channel
myocyte Ca2+ signaling . . . . . . . . . . . . . 107 open probability to explain the residual
18.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107 Ca2+ influx . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 160
18.2 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108
18.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109 22 Ion channels in the human red
18.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114 blood cell membrane: Their further
18.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 118 investigation and physiological
relevance . . . . . . . . . . . . . . . . . . . . . . . . . . 162
19 Remodelling of Ca2+ handling 22.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
organelles in adult rat ventricular 22.2 Experimental . . . . . . . . . . . . . . . . . . . . . . . . . . . 162
myocytes during longterm culture . . 121 22.3 Results and discussion . . . . . . . . . . . . . . . . . . 163
19.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121 22.4 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
19.2 Material and methods . . . . . . . . . . . . . . . . . . 122
19.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123 23 Prostaglandin E activates channel-
2
19.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130 mediated calcium entry in human
19.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133 erythrocytes: An indication for a
blood clot formation supporting
process . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
23.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 166
23.2 Material and methods . . . . . . . . . . . . . . . . . . 166
XI
Content
23.3 Results and discussion . . . . . . . . . . . . . . . . . . 167 28.5 Research perspective . . . . . . . . . . . . . . . . . . . 230
23.4 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169 28.6 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 231
24 Functional NMDA receptors in Subject Index . . . . . . . . . . . . . . . . . . . . . . 235
rat erythrocytes . . . . . . . . . . . . . . . . . . . . 171
24.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 171
24.2 Materials and Methods . . . . . . . . . . . . . . . . . 172
24.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 175
24.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 182
24.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
25 Stimulation of human red blood
cells leads to Ca2+-mediated
intercellular adhesion . . . . . . . . . . . . . . . 187
25.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
25.2 Materials and methods . . . . . . . . . . . . . . . . . 188
25.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
25.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 195
25.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197
26 Lysophospatidic acid induced red
blood cell aggregation in vitro . . . . . . 200
26.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 200
26.2 Material and methods . . . . . . . . . . . . . . . . . . 201
26.3 Results and discussion . . . . . . . . . . . . . . . . . . 202
26.4 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 206
26.5 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 208
27 Regulation of phosphatidylserine
exposure in red blood cells . . . . . . . . . . 210
27.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 210
27.2 Materials and Methods . . . . . . . . . . . . . . . . . 211
27.3 Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 213
27.4 Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 218
27.5 Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
27.6 References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 220
28 Cation Channels in Erythrocytes -
Historical and Future Perspective . . . . 223
28.1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . 223
28.2 The discovery of cation channels in
erythrocytes: from flux-measurements
to electrophysiology . . . . . . . . . . . . . . . . . . . . 224
28.3 Classification and properties of
channels achieved by the patch-clamp
technique . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 226
28.4 Towards the molecular idendity of the
channels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 229
I
Summary
Chapter 1 Introduction – 2
Chapter 2 Direct evidence - the digital approach – 3
Chapter 3 Fluorescence-based visualisation – 4
Chapter 4 Aequorin-based measurements – 14
Chapter 5 Measurement of calcium transport across
membranes – 15
Chapter 6 Molecular biology based approaches – 17
Chapter 7 Manipulation of calcium – 19
Chapter 8 Calcium-induced function – 21
Chapter 9 Calcium signalling in cardiac myocytes – 24
Chapter 10 Calcium signalling in red blood cells – 26
Chapter 11 Perspective – 29