Stefan Dhein Friedrich Wilhelm Mohr Mario Delmar. Practical Methods in Cardiovascular Research

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1 Stefan Dhein Friedrich Wilhelm Mohr Mario Delmar Practical Methods in Cardiovascular Research

2 Stefan Dhein (Editor) Friedrich Wilhelm Mohr (Editor) Mario Delmar (Editor) Practical Methods in Cardiovascular Research With 33 Tables and 290 Figures

3 Prof. Dr. Stefan Dhein Prof. Dr. Friedrich Wilhelm Mohr Mario Delmar, MD PhD Department of Pharmacology SUNY Upstate Medical University 766 Irving Avenue Syracuse NY Library of Congress Control Number: ISBN Springer Berlin Heidelberg New York This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilm or in any other way, in storage in data banks. Duplications of this publication or parts thereof is permitted only under the provision of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer-Verlag. Violations are liable for prosecution under the German Copyright Law. Springer is a part of Springer Science+Business Media springeronline.com Springer-Verlag Berlin Heidelberg 2005 Printed in The use of general descriptive names, registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Product liability: The publisher cannot guarantee the accuracy of any information about dosage and application thereof contained in this book. In every individual case the user must check such information by consulting the relevant literature. Editor: Dr. Thomas Mager, Heidelberg Development Editor: Natasja Sheriff, Heidelberg Production: Frank Krabbes, Heidelberg Cover Design: deblik, Berlin Page makeup: Hilger VerlagsService, Heidelberg Printing: Krips BV, Meppel, Niederlande Printed on acid-free paper SPIN: /

4 Preface V 0.00 Preface Cardiovascular disease is among the most common causes of death in Western industrialized countries. Although a number of diagnostic and therapeutic possibilities exist today, many problems remain unsolved. This implies the urgent need for future cardiovascular research. The present book gives an overview of a number of available techniques most often used in cardiovascular sciences, especially for investigators coming from other fields of research. The book is divided into three sections. In the first part, the focus is on techniques used for in vivo experimentation. This field has regained great interest, given the availability of knock-out models which now await in-vivo testing. The book describes the most commonly used procedures for anesthesia and analgesia in laboratory animals as well as classical isolated organ techniques, including electrophysiological methods now again in common use. In the second part of the book, we move from the organ and tissue level to the cellular domain. Topics covered include electrophysiology, histology and cell culture techniques as well as biochemical methods such as HPLC, protein biochemistry and binding assays. This part closes with a brief chapter on how to register and analyze concentration response curves giving briefly the mathematics and basic analysis techniques. In the final part, the focus narrows further into molecular procedures such as surface plasmon resonance, nuclear magnetic resonance, RNA methods and DNA arrays. Finally, a chapter is devoted to the molecular aspects of transgenic mice technology as well as an overview of procedures and regulations applicable to housing and transfer of transgenic animals. The novelty of this book is in the description of practical protocols and troubleshooting procedures. It is our intention to provide a basic guide for investigators interested in applying these techniques in their own laboratories. The book also aims to close the gap between the integrative in-vivo and organ-based methods and the reductionist cellular and molecular approaches. We wish all researchers every success with their experiments and that this book may help them start their way toward new frontiers. Stefan Dhein Mario Delmar Friedrich Wilhelm Mohr

5 Table of Contents VII 0.00 Table of Contents In-Vivo Techniques 1 Models of Cardiovascular Diseases 1.1 Anaesthesia (Inhalative or i.v. Narcosis) of Laboratory Animals Katja Schneider 1.2 Extracorporeal Circulation Cris W. Ullmann 1.3 Open-chest Models of Acute Myocardial Ischemia and Reperfusion Kai Zacharowski, Thomas Hohlfeld and Ulrich K.M. Decking 1.4 Models of Chronic Ischemia and Infarction Nicolas Doll, Heike Aupperle, Fabian Stahl and Thomas Walther 1.5 Models on Left Ventricular Hypertrophy (LVH) Thomas Walther and Volkmar Falk 1.6 Experimental Models of Heart Failure Volkmar Falk, Jens Garbade and Thomas Walther 1.7 In-Vivo Models of Arrhythmias: a Canine Model of Sudden Cardiac Death George E. Billman 1.8 In-Vivo Models of Atrial Fibrillation Ulrich Schotten, Yuri Blaauw and Maurits Allessie In-Vitro Techniques 2 Isolated Organs 2.1 The Langendorff Heart Stefan Dhein 2.2 The Working Heart Marc W. Merx and Jürgen Schrader 2.3 Isolated Papillary Muscles Stefan Dhein 2.4 Isolated Vessels Rudolf Schubert

6 0 VIII Table of Contents 3 Electrophysiological Techniques 3.1 Optical Techniques for the Recording of Action Potentials Helmut A. Tritthart 3.2 Recording Action Potentials Using Voltage-Sensitive Dyes Vladimir G. Fast 3.3 Epicardial Mapping in Isolated Hearts Use in Safety Pharmacology, Analysis of Torsade de Pointes Arrhythmia and Ischemia-Reperfusion-Related Arrhythmia Stefan Dhein 3.4 Voltage-Clamp and Patch-Clamp Techniques Hans Reiner Polder, Martin Weskamp, Klaus Linz, Rainer Meyer 3.5 L-Type Calcium Channel Recording Uta C. Hoppe, Mathias C. Brandt, Guido Michels and Michael Lindner 3.6 Recording Cardiac Potassium Currents with the Whole-Cell Voltage Clamp Technique Erich Wettwer and Ursula Ravens 3.7 Recording the Pacemaker Current If Uta C. Hoppe, Fikret Er and Mathias C. Brandt 3.8 Recording Gap Junction Currents Ardawan J. Rastan and Stefan Dhein 3.9 Recording Monophasic Action Potentials Paulus Kirchhof, Larissa Fabritz and Michael R. Franz 3.10 Heart on a Chip Extracellular Multielectrode Recordings from Cardiac Myocytes in Vitro Ulrich Egert and Thomas Meyer 4 Histological Techniques 4.1 Immunohistochemistry for Structural and Functional Analysis in Cardiovascular Research Wilhelm Bloch, Yüksel Korkmaz and Dirk Steinritz 4.2 Classical Histological Staining Procedures in Cardiovascular Research Wilhelm Bloch and Yüksel Korkmaz 4.3 Practical Use of in situ Hybridisation and RT in situ PCR in Cardiovascular Research Yüksel Korkmaz, Dirk Steinritz and Wilhelm Bloch 4.4 Practical Use of Transmission Electron Microscopy for Analysis of Structures and Molecules in Cardiovascular Research Wilhelm Bloch, Christian Hoffmann, Eveline Janssen and Yüksel Korkmaz 4.5 DAF Technique for Real-Time NO Imaging in the Human Myocardium Dirk Steinritz and Wilhelm Bloch

7 Table of Contents IX Cell Culture Techniques 5.1 Isolation and Culture of Adult Ventricular Cardiomyocytes Klaus-Dieter Schlüter and Hans Michael Piper 5.2 Culture of Neonatal Cardiomyocytes Aida Salameh and Stefan Dhein 5.3 Culture of Embryoid Bodies Cornelia Gissel, Dirk Nierhoff, Bernd Fleischmann, Jürgen Hescheler and Agapios Sachinidis 5.4 Culturing and Differentiation of Embryonic and Adult Stem Cells for Heart Research and Transplantation Therapy Marcel AG van der Heyden and Henk Rozemuller 5.5 Preparation of Endothelial Cells from Micro- and Macrovascular Origin Saskia C. Peters, Anna Reis and Thomas Noll 5.6 In Vitro Cultivation of Vascular Smooth Muscle Cells Daniel G. Sedding and Ruediger C. Braun-Dullaeus 5.7 Engineering Heart Tissue for In Vitro and In Vivo Studies Wolfram-Hubertus Zimmermann, Ivan Melnychenko, Michael Didié, Ali El-Armouche and Thomas Eschenhagen 6 Biochemical and Analytical Techniques 6.1 High-Performance Liquid Chromatography (HPLC) Veronika R. Meyer 6.2 Protein Biochemistry Aida Salameh, Stefan Lehr, Jürgen Weiss and Peter Rösen 6.3 Receptor and Binding Studies Peter Hein, Martin C. Michel, Kirsten Leineweber, Thomas Wieland, Nina Wettschureck and Stefan Offermanns 6.4 Recording and Analysing Concentration-Response Curves Stefan Dhein 6.5 Measurement of NO and Endothelial Function in Cardiovascular Research Renate Roesen, Anke Rosenkranz, Dirk Taubert and Reinhard Berkels 6.6 Biomechanical Stimulation of Vascular Cells In Vitro Henning Morawietz and Andreas Schubert 6.7 Measurement of Function and Regulation of Adrenergic Receptors Marc Brede, Melanie Philipp and Lutz Hein 6.8 Measurement of Function and Regulation of Muscarinic Acetylcholine Receptors Björn Kaiser and Chris J. van Koppen 6.9 Cytometry in Cardiovascular Research Attila Tárnok

8 0 X Table of Contents Molecular Biology in Cardiovascular Research 7 Molecular Biology Techniques 7.1 Real-Time PCR Analysis of Cardiac Samples Dalton A. Foster and Steven M. Taffet 7.2 Use of cdna Arrays to Explore Gene Expression in Genetically Manipulated Mice and Cell Lines Dumitru A. Iacobas, Sandra Iacobas and David C. Spray 7.3 Using Antibody Arrays to Detect Protein-Protein Interactions Heather S. Duffy, Ionela Iacobas, David C. Spray and Anthony W. Ashton 7.4 Surface Plasmon Resonance as a Method to Study the Kinetics and Amplitude of Protein-Protein Binding Brian D. Lang, Mario Delmar and Wanda Coombs 7.5 How to Solve a Protein Structure by Nuclear Magnetic Resonance The Connexin43 Carboxyl Terminal Domain Paul L. Sorgen 7.6 Managing a Transgenic Mouse Colony: A Guide for the Novice Robert H. Quinn and Karen L. Vikstrom Appendix Colour Plates Index

9 List of Abbreviations XI 0.00 List of Abbreviations AP 1D 2D 3D 5-HT α 1 AR AADC ABC AC Ach ACT AEC AF AMI Ao APHIS Ara-C ARI AT-1 AT-2 B0 β3-ar BCIP BCL BDM BM BrdU BSA BSA BSS BTP action potential one-dimensional two-dimensional three-dimensional serotonin α 1 -Adrenoceptor aromatic L-amino acid decarboxylase avidin-biotin peroxidase complex adenylyl cyclase acetylcholine activated clotting time 3-amino-9-ethylcarbazole atrial fibrillation acute myocardial infarction Aorta Agriculture Animal and Plant Health Inspection Service cytosine-β-d-arabinofuranoside activation-recovery-interval angiotensin type 1 receptor angiotensin type 2 receptor the static magnetic field β3-adrenoceptors 5-bromo-4-chloro-3- indolylphosphate basic cycle length 2,3-butanedione-monoxime bone marrow 5-bromo-2-desoxyuridine bovine serum albumin biotin-streptavidin balanced salt saline breakthrough-points COMT COS cells CPB CS CSD CV Cx43 CY DA DAB DAFs DAF-T Daun DβH dd DDSA DEPC DMEM DMEM-F12 DMP DMSO Dox dp/dt max dp/dt min dp/dt min DSEVC DSS DTT catechol-o-methyltransferase kidney cells from the monkey Cercopithecus aethiops cardiopulmonary bypass cell signaling cell cycle-shape-differentiation superior caval vein connexin43 cytoskeletal diacetate diaminobenzidine tetrahydrochloride diaminofluoresceins triazole-fluoresceins daunorubicin dopamine β-hydroxylase double-distilled dodecenyl succinic anhydride diethylpyrocarbonate Dulbecco s modified Eagle medium Dulbecco s Modified Medium tri(dimethylaminomethyl)phenol dimethylsulfoxide doxorubicin maximum rate of left ventricular pressure development maximum relaxation velocity minimum rate of pressure development discontinuous single electrode voltage clamp amplifier Dahl salt sensitive rat dithiothreitol C CABG CARB CARD CCD CDC CF CHO CK CK/CKMB CK-MB CN CNBD CNS CO capacitance coronary artery bypass grafting carbachol catalyzed reporter deposition charge-coupled device Centers for Disease Control coronary flow Chinese hamster ovary cells creatine kinase ccreatine kinase/cardiospecific CK cardiospecific CK cellulose nitrate cyclic nucleotide-binding domain crystallography & NMR systems cardiac output EBNA-LP EBs EC EC ECG ECL HRP EDC EDP EDTA EGF EGFP EnMet Epstein-Barr virus nuclear antigenleader protein embryoid bodies embryonal carcinoma endothelial cells electrocardiogram enhanced chemiluminescence horseradish peroxidase 1-Ethyl 3-3-dimethylaminopropyl carbodiimide end-diastolic pressure ethylenediaminetetraacetate epidermal growth factor enhanced green fluorescent protein Energy and Metabolism

10 0 XII List of Abbreviations IUPAC IV JAE Kv-channels LA LAD LCX LDH LIF LV LVDP LVEF LVP MACS MALDI MAO MAP MAP MAP MAPC Mb MBF MEA MEF MEM MHC MI MLP MNC MOI MRI MS MS/MS MSC NA NAT NBT NCS NHS NMR NO The International Union of Pure and Applied Chemistry intravenous junction-adhesion-extracellular matrix voltage-gated potassium channels left atrium left anterior descending coronary artery left circumflex artery lactate dehydrogenase leukemia inhibitory factor left ventricle left ventricular developed pressure left ventricular ejection fraction left ventricular pressure enos endothelial nitric oxide synthase EPC endothelial progenitor cells Epi epirubicin ERP effective refractory period ES embryonic stem ESI electrospray-ionisation ESTs expressed sequence tags ET-1 endothelin-1 ET A R endothelin-1 receptor subtype A EtOH ethanol FACS fluorescence activated cell sorter FCS foetal calf serum FISH fluorescence in situ hybridisation FITC- fluorescein-isothiocyanate- FOC force of contraction FP field potentials Gab Grb-associated binder GAP GTPase-activating protein GFP green fluorescent protein GPCR G protein coupled receptor HBSS Hank s balanced salt solution HCN channels hyperpolarisation-activated cyclic nucleotide-gated cation channels HE hematoxylin-eosin HEK human embryonic kidney cells HEPA-filter high efficiency particulate air filter HEPES-Na 2-[4-(2-Hydroxyethyl)-1-piperazinyl]- ethanesulphonic acid sodium salt HR Heart rate HRP horseradish peroxidase HSC haematopoietic stem cells HSQC 1 H 15 N-heteronuclear single quantum coherence HW heart weight IACUC Institutional Animal Care and Use Committees IC intracoronary ICM inner cell mass IMAC immobilized metal ion affinity chromatography IMDM Iscove s modified Dulbecco s medium IP intraperitoneal IPPV intermittent positive pressure ventilation ITO indium-tin oxide magnetic activated cell sorting matrix-assisted-laser-desorptionionisation monoamine oxidase mean arterial pressure mitogen activated protein monophasic action potentials multipotent adult progenitor cell myoglobin myocardial blood flow microelectrode arrays mouse embryonic fibroblast non-essential amino acids myosin heavy chain myocardial infarction muscle LIM-domain protein mononucleated cells multiplicity of infection magnetic resonance imaging mass spectrometry tandem mass spectrometry mesenchymal stem cells noradrenalin nordadrenaline transporter nitro blue tetrazolium chloride newborn calf serum N-hydroxysuccinimide nuclear magnetic resonance nitric oxide

11 List of Abbreviations XIII 0.00 NOE NOS Ntg OG PAP PAR1 PBS PCNA PCR PD PDA PDEA PDGF PE PE PEEP PEG PEI PenStrep PET PFA P-I PIPES PKA PL PLC-β PLD PMMA PNMT PPM PQ PSD PTCA PTK PTP PVDF QTc R RA RCA nuclear overhauser effect NO-synthases non-transgenic organelle genetics peroxidase anti-peroxidase thrombin activated protease receptor type 1 phosphate buffered saline proliferating nuclear antigen polymerase chain reaction proportional distance photodiode arrays (2-(2-pyridinylthio)ethanamine hydrochloride) platelet derived growth factor phycoerythrin phenylephrine positive end expiratory pressure polyethylene glycol polyethylenimine penicillin/streptomycin solution positron emission para-formaldehyde proportional integral piperazine-n -N -bis[2-ethanesulfonic acid camp-dependent protein kinase intrapleural β-isoforms of phospholipase C phospholipase D polymethylmethacrylate phenylethanolamine-n-methyltransferase parts per million atrioventricular conduction time post source decay percutaneous transluminal coronary angioplasty protein tyrosine kinase protein tyrosine phosphatase polyvinylidendifluoride frequency corrected QT time resistance right atrium right coronary artery RCX circumflex artery REV relative estimated (transcription) variability RGS regulators of G protein signalling ROS reactive oxygen species RP reverse-phase rpm revolutions per minute RT room temperature RT reverse transcription RT 1/2 relaxation halftime RU response units S/N signal-to-noise ratio SAN sino-atrial node SC subcutaneous SCX strong cation exchange SECV single electrode voltage clamp sgc soluble guanyl cyclase SHR spontaneously hypertensive rats SMC smooth muscle cell SM-MHC smooth muscle myosin heavy chain SNR signal-to-noise ratio SO Symmetrical Optimum SP side population SP systolic pressure SPF specific-pathogen-free SPR surface plasmon resonance SSC standard saline citrate synthase L-NNA T tesla T1 transport into the cell T2 transport within the cell TAT total activation time TB Tris buffer TBS Tris-buffered saline TBST 150 mm NaCl, 25 mm Tris-HCl, 0.05% Tween20, ph 7.5 TD trypsin-diluted TdP torsade de pointes TE expiratory ventilation time TEA tetraethylammonium aspartate TEVC two-electrode voltage clamp Tg transgenic TH tyrosine hydroxylase TI inspiratory ventilation time TMR transmyocardial laser revascularisation TPP time to peak pressure TR transcriptional

12 0 XIV List of Abbreviations Tris TSA TSP tta TTX trishydroxymethylaminomethane tyramide signal amplification 3-(Trimethylsilyl)- Propionic acid-d4 tetracycline-controlled transactivation tetrodotoxin VMAT V rev VT VWF vesicular monoamine transporter reversal potential transverse ventricular propagation velocity von Willebrand factor USDA United States Department of Agriculture WPW WT Wolff-Parkinson-White syndrome wildtype VDCC V L voltage dependent calcium channel longitudinal ventricular propagation velocity YAC yeast artificial chromosome

13 List of Contributors XV 0.00 List of Contributors Allessie, Maurits A. University Maastricht Department of Physiology PO Box MD Maastricht The Netherlands Ashton, Anthony W. Albert Einstein College of Medicine Department of Neuroscience Bronx, New York Aupperle, Heike Berkels, Reinhard Gleueler Straße 24 Billman, George E. The Ohio State University Department of Physiology and Cell Biology 304 Hamilton Hall 1645 Neil Ave. Columbus OH Blaauw, Yuri University Maastricht Department of Physiology PO Box MD Maastricht The Netherlands Bloch, Wilhelm Deutsche Sporthochschule Köln Institut für Kreislaufforschung und Sportmedizin Carl-Diem-Weg Köln Brandt, Mathias C. Klinik für Innere Medizin III Joseph-Stelzmann Straße Köln Braun-Dullaeus, Ruediger C. Technische Universität Dresden Klinik für Innere Medizin Fetscherstraße Dresden Brede, Marc Bayerische Julius-Maximilians- Universität Würzburg und Toxikologie Versbacher Straße Würzburg Coombs, Wanda SUNY Upstate Medical University Department of Pharmacology 766 Irving Ave. Syracuse NY Decking, Ulrich Heinrich-Heine-Universität Universitätsklinikum Institut für Herz- und Kreislaufphysiologie Universitätsstraße Düsseldorf Delmar, Mario SUNY Upstate Medical University Department of Pharmacology 766 Irving Ave. Syracuse NY Dhein, Stefan Didié, Michael Universitätsklinikum Hamburg- Eppendorf Zentrum für Experimentelle Medizin Institut für Experimentelle und Klinische Pharmakologie Martinistraße Hamburg Doll, Nicolas Duffy, Heather S. Albert Einstein College of Medicine Department of Neuroscience Bronx, New York Egert, Ulrich Universität Freiburg Neurobiologie und Biophysik Institut für Biologie III Schänzlestraße Freiburg

14 0 XVI List of Contributors El-Armouche, Ali Universitätsklinikum Hamburg- Eppendorf Zentrum für Experimentelle Medizin Institut für Experimentelle und Klinische Pharmakologie Martinistraße Hamburg Er, Fikret Klinik für Innere Medizin III Joseph-Stelzmann-Straße Köln Eschenhagen, Thomas Universitätsklinikum Hamburg- Eppendorf Zentrum für Experimentelle Medizin Institut für Experimentelle und Klinische Pharmakologie Martinistraße Hamburg Fabritz, Larissa Universitätsklinikum Münster Medizinische Klinik und Poliklinik C Kardiologie und Angiologie und Institut für Arterioskleroseforschung Albert-Schweitzer-Straße Münster Falk, Volkmar Fast, Vladimir G. University of Alabama at Birmingham 1670 University Blvd., VH B126 Birmingham, AL Fleischmann, Bernd Rheinische-Friedrich-Wilhelms- Universität Bonn Institut für Physiologie I Argelander Straße 2a Bonn Franz, Michael Georgetown University and Veterans Administrations Medical Centers Departments of Cardiology and Pharmacology 50 Irving Street, NW Washington DC Foster, Dalton A. SUNY Upstate Medical University Department of Microbiology and Immunology 750 E. Adams St Syracuse, NY Garbade, Jens Gissel, Cornelia Institut für Neurophysiologie Robert-Koch-Straße 39 Hein, Lutz Bayerische Julius-Maximilians- Universität Würzburg und Toxikologie Versbacher Straße Würzburg Hein, Peter Bayerische Julius-Maximilians- Universität Würzburg und Toxikologie Versbacher Straße Würzburg Hescheler, Jürgen Institut für Neurophysiologie Robert-Koch-Straße 39 van der Heyden, Marcel A.G. University Medical Center Department of Medical Physiology Universiteitsweg CG Utrecht The Netherlands Hoffmann, Christian Zentrum Anatomie Institut I für Anatomie Joseph-Stelzmann-Straße 9 Hohlfeld, Thomas Universität Düsseldorf Moorenstraße Düsseldorf

15 List of Contributors XVII 0.00 Hoppe, Uta C. Klinik für Innere Medizin III Joseph-Stelzmann Str Köln Iacobas, Dumitru A. Albert Einstein College of Medicine Department of Neuroscience Bronx, New York Iacobas, Ionela Albert Einstein College of Medicine Department of Neuroscience Bronx, New York Iacobas, Sandra Albert Einstein College of Medicine Department of Neuroscience Bronx, New York Janssen, Eveline Zentrum Anatomie Institut I für Anatomie Joseph-Stelzmann-Straße 9 Kaiser, Björn Universitätsklinikum Essen Hufelandstraße Essen Kirchhof, Paulus Universitätsklinikum Münster Medizinische Klinik und Poliklinik C Albert-Schweitzer-Straße Münster van Koppen, Chris J. Universitätsklinikum Essen Hufelandstraße Essen Korkmaz, Yüksel Zentrum Anatomie Institut I für Anatomie Joseph Stelzmann Straße 9 Lang, Brian D. Biacore Inc., Piscataway, New Jersey Lehr, Stefan Heinrich-Heine-Universität Düsseldorf Institut für Klinische Biochemie und Pathobiochemie Diabetes Forschungszentrum Leibniz-Zentrum für Diabetes- Forschung Auf m Hennekamp Düsseldorf Leineweber, Kirsten Universitätsklinikum Essen Institut für Pathophysiologie Hufelandstraße Essen Lindner, Michael Klinik für Innere Medizin III Joseph-Stelzmann-Straße Köln Linz, Klaus Grünenthal GmbH Safety Pharmacology Zieglerstraße Aachen Melnychenko, Ivan Universitätsklinikum Hamburg- Eppendorf Zentrum für Experimentelle Medizin Institut für Experimentelle und Klinische Pharmakologie Martinistraße Hamburg Merx, Marc W. Universitätsklinikum Aachen Medizinische Klinik 1 Kardiologie und Pneumologie Pauwelstraße Aachen Meyer, Rainer Rheinische-Friedrich-Wilhelms- Universität Bonn Institut für Physiologie II Wilhelmstraße Bonn Meyer, Thomas Multi Channel Systems MCS GmbH Aspenhaustraße Reutlingen Meyer, Veronika R. EMPA St. Gallen Eidgenössische Materialprüfungs- und Forschungsanstalt Metrologie in der Chemie Lerchenfeldstrasse St. Gallen Switzerland

16 0 XVIII List of Contributors Michels, Guido Klinik für Innere Medizin III Joseph-Stelzmann-Straße Köln Michel, Martin C. University of Amsterdam Academic Medical Center Dept. Pharmacology & Pharmacotherapy Meibergdreef AZ Amsterdam The Netherlands Morawietz, Henning Technische Universität Dresden Medizinische Klinik und Poliklinik III Bereich Gefäßendothel/ Mikrozirkulation Fetscherstraße Dresden Nierhoff, Dirk Institut für Neurophysiologie Robert-Koch-Straße 39 Noll, Thomas Justus-Liebig-Universität Giessen Physiologisches Institut Aulweg Giessen Offermanns, Stefan Universität Heidelberg Pharmakologisches Institut Im Neuenheimer Feld Heidelberg Peters, Saskia C. Justus-Liebig-Universität Giessen Physiologisches Institut Aulweg Giessen Philipp, Melanie Universität Zürich Institut für Zoologie Winterthurerstrasse Zürich Switzerland Piper, Hans Michael Justus-Liebig-Universität Giessen Physiologisches Institut Aulweg Giessen Polder, Hans Reiner npi electronic GmbH Hauptstraße Tamm Quinn, Robert H. SUNY Upstate Medical University Department of Laboratory Animal Resources 750 East Adams Street Syracuse, NY Rastan, Ardawan J. Ravens, Ursula Technische Universität Dresden und Toxikologie Fetscherstraße Dresden Reis, Anna Justus-Liebig-Universität Giessen Physiologisches Institut Aulweg Giessen Rösen, Peter Heinrich-Heine-Universität Düsseldorf Institut für Klinische Biochemie und Pathobiochemie Leibniz-Zentrum für Diabetes- Forschung Auf m Hennekamp Düsseldorf Rösen, Renate Gleueler Straße 24 Rosenkranz, Anke Gleueler Straße 24 Rozemuller, Henk Jordan Laboratory University Medical Center Department of Heamatology Universiteitsweg CG Utrecht The Netherlands Sachinidis, Agapios Institut für Neurophysiologie Robert-Koch-Straße 39

17 List of Contributors XIX 0.00 Salameh, Aida Klinik I für Innere Medizin Johannisallee Leipzig Schlüter, Klaus-Dieter Justus-Liebig-Universität Giessen Physiologisches Institut Aulweg Giessen Schneider, Katja Schotten, Ulrich University Maastricht Department of Physiology PO Box MD Maastricht The Netherlands Schrader, Jürgen Heinrich-Heine-Universität Düsseldorf Institut für Physiologie Postfach Düsseldorf Schubert, Andreas Russenstraße 19 Schubert, Rudolf Universität Rostock Institut für Physiologie Postfach Rostock Sedding, Daniel G. Justus-Liebig-Universität Giessen Medizinische Klinik I Innere Medizin/Kardiologie Klinikstraße Giessen Sorgen, Paul. L. University of Nebraska Medical Center Department of Biochemistry and Molecular Biology Omaha, NE Spray, David C. Albert Einstein College of Medicine Department of Neuroscience Bronx, New York Stahl, Fabian Steinritz, Dirk Zentrum Anatomie Institut I für Anatomie Joseph-Stelzmann-Straße 9 Taffet, Steven M. SUNY Upstate Medical University Department of Microbiology and Immunology 750 E. Adams St Syracuse, NY Tárnok, Attila Klinik für Kinderkardiologie Taubert, Dirk Gleueler Straße 24 Tritthart, Helmut A. Karl-Franzens-Universität Graz Institut für Medizinische Physik und Biophysik Harrachgasse Graz Austria Ullmann, Cris W. Vikstrom, Karen L. SUNY Upstate Medical University Department of Laboratory Animal Resources 750 East Adams Street Syracuse, NY Walther, Thomas

18 0 XX List of Contributors Weiß, Jürgen Heinrich-Heine-Universität Düsseldorf Institut für Klinische Biochemie und Pathobiochemie Leibniz-Zentrum für Diabetes- Forschung Auf m Hennekamp Düsseldorf Weskamp, Martin npi electronic GmbH, Hauptstraße 96, Tamm Wettschureck, Nina Universität Heidelberg Pharmakologisches Institut Im Neuenheimer Feld Heidelberg Wettwer, Erich Technische Universität Dresden und Toxikologie Fetscherstraße Dresden Wieland, Thomas Universität Heidelberg Fakultät für Klinische Medizin Mannheim und Toxikologie Maybachstraße Mannheim Zacharowski, Kai Heinrich-Heine-Universität Düsseldorf Universitätsklinikum Klinik für Anästhesiologie und Intensivmedizin Moorenstraße Düsseldorf Zimmermann, Wolfram- Hubertus Universitätsklinikum Hamburg- Eppendorf Zentrum für Experimentelle Medizin Institut für Experimentelle und Klinische Pharmakologie Martinistraße Hamburg