Manual of Research Techniques in Cardiovascular Medicine

Edited by Hossein Ardehali MD, PhD, FAHA, Associate Professor of Medicine, Molecular Pharmacology and BiologicalChemistryFeinberg Cardiovascular Research InstituteNorthwestern UniversityFeinberg School ofMedicine, Chicago, IL USARoberto Bolli MDProfessor of Medicine Chief, Division of Cardiovascular MedicineDirector, Institute of Molecular CardiologyUniversity of LouisvilleLouisville, KY USADouglas W. Losordo MDVice President New Therapeutic DevelopmentBaxter Healthcare CorporationDeerfield, IL USA

• List of Contributors ixPreface xixAbout the Companion Website xxPart 1 Electrophysiology1 Measurement of calcium transient ex vivo 3Kenneth R. Laurita and Bradley N. Plummer2 Confocal imaging of intracellular calcium cycling in isolated cardiac myocytes 12Søren Grubb, J. Andrew Wasserstrom, and Gary L. Aistrup3 Generating a large animal model of persistent atrial fi brillation 20Raphaël P. Martins and José Jalife4 Confocal imaging of intracellular calcium cycling in the intact heart 32Neha Singh, Manvinder Kumar, James E. Kelly, Gary L. Aistrup, and J. Andrew Wasserstrom5 Recording and measurement of action potentials 41Kenneth S. Ginsburg and Donald M. Bers6 Patch-clamp recordings from isolated cardiac myocytes 50Kai-Chien Yang, Wei Wang, and Jeanne M. Nerbonne7 Optical mapping of the heart 60Di Lang, Sarah Gutbrod, Jacob Laughner, and Igor R. Efi movPart 2 Isolation and Maintenance of Primary Stem Cells8 Isolation of colony-forming endothelial progenitor cells 71Haruchika Masuda and Takayuki Asahara9 Cardiac resident stem cells 87João Ferreira-Martins, Fumihiro Sanada, and Marcello Rota10 Cardiospheres 95Rachel Ruckdeschel Smith11 Mesenchymal stem cells 104Jose S. Da Silva and Joshua M. Hare12 Generation and differentiation of human iPS cells 110Sebastian Diecke, Lei Ye, Sophia Zhang, and Jianyi Zhang13 Isolation of neonatal and adult rat cardiomyocytes 117Md. Abdur Razzaque and Jeffrey Robbins14 Isolation and culture of vascular smooth muscle cells 125Milton Hamblin, Lin Chang, and Y. Eugene Chen15 Isolation and culture of cardiac endothelial cells 131Asish K. Ghosh, Joseph W. Covington, and Douglas E. Vaughan16 Isolation and culture of cardiac fi broblasts 140Asish K. Ghosh, Joseph W. Covington and Douglas E. Vaughan17 Murine bone marrow transplantation model 146Prasanna Krishnamurthy, Suresh Kumar Verma, and Raj Kishore18 In vitro differentiation and expansion of vascular endothelial cells derived from mouse embryonic stem cells 149Anees Fatima, Carey Nassano-Miller, and Tsutomu KumePart 3 Manipulation of the Heart and Vessels in Vivo and ex Vivo19 Coronary ligation 157Alexander R. Mackie and Hossein Ardehali20 Transverse aortic constriction: a model to study heart failure in small animals 164Suresh Kumar Verma, Prasanna Krishnamurthy, and Raj Kishore21 Pharmacological models of hypertrophy and failure 170Angela C. deAlmeida, Tariq Hamid, and Sumanth D. Prabhu22 Hindlimb ischemia 177Jerry C. Lee, Ngan F. Huang, and John P. Cooke23 The Langendorff preparation 187Hugh Clements-Jewery and Michael J. Curtis24 Myocarditis and other immunological models of cardiac disease 197Daniela C¡ iháková and Noel R. Rose25 Models of pacing-induced heart failure 203James A. Shuman, Rupak Mukherjee, and Francis G. Spinale26 Porcine myocardial ischemia models 215Xian-Liang Tang and Roberto Bolli27 Angiogenesis assays 225Susmita Sahoo and Douglas W. Losordo28 Immunohistochemical analysis of cardiac tissue 232Barbara Ogórek, Donato Cappetta, and Jan Kajstura29 A murine model of cardiac arrest by exsanguination 237Guangming Cheng, Yiru Guo, Harold K. Elias, Carrie M. Quinn, Arash Davani, Yanjuan Yang, Magdy Girgis, Roberto Bolli, and Buddhadeb DawnPart 4 Small Animal Imaging30 Blood pressure, telemetry, and vascular measurements in the rodent model 245Robert S. Danziger31 The setting: imaging conscious, sedated, or anesthetized rodents 250Gene H. Kim and Roberto M. Lang32 Echocardiography: standard techniques (M-mode, two-dimensional imaging, and Doppler) 254Gene H. Kim, Lauren Beussink-Nelson, Sanjiv J. Shah, and Roberto M. Lang33 Echocardiography: advanced techniques (tissue Doppler, speckle tracking, and three-dimensional imaging) 275Lauren Beussink-Nelson, Gene H. Kim, Roberto M. Lang, and Sanjiv J. Shah34 In vivo tomographic cardiac imaging: positron emission tomography and magnetic resonance imaging 287Bruno C. Huber, Patricia K. Nguyen, and Joseph C. Wu35 In vivo hemodynamics 295Alexander R. Mackie, Kyle K. Henderson, Sol Misener, and Hossein ArdehaliPart 5 Metabolism, Mitochondria, and Cell Death36 Fractionation of cardiomyocytes and isolation of mitochondria 307Christopher P. Baines37 Assessment of glucose and fatty acid metabolism ex vivo 313Darrell D. Belke and E. Dale Abel38 Quantification and characterization of atherosclerotic lesions in mice 320Abhinav Agarwal, Millicent G. Winner, Srinivas D. Sithu, and Sanjay Srivastava39 Assessment of cell death in the heart 332Russell S. Whelan, Klitos Konstantinidis, and Richard N. Kitsis40 Assessment of mitochondrial function in isolated cells 343Amy K. Rines and Hossein Ardehali41 Multinuclear NMR spectroscopy of myocardial energetics and substrate utilization in isolated perfused mouse hearts 351Stephen C. Kolwicz, Jr. and Rong Tian42 Measurement of reactive oxygen species in cardiovascular disease 359Mahmood Khan, Fatemat Hassan, Sashwati Roy, and Chandan K. Sen43 Assessing autophagy 371Roberta A. Gottlieb44 Assessment of cardiomyocyte size 378A. Martin Gerdes and Alessandro PingitorePart 6 Manipulation of Gene Expression in Vitro and in Vivo45 Generation of Cre-loxP mouse models for conditional knockout and overexpression of genes in various heart cells 389Marisa Z. Jackson and Warren G. Tourtellotte46 Modulation of myocardial genes via use of adenoviral vectors and RNA interference approaches 397Qianhong Li47 Overexpression and downregulation of proteins in vitro 407Marina Bayeva and Hossein Ardehali48 In vivo microRNA studies 416Eva van RooijPart 7 Model Systems49 Vascular and cardiac studies in zebrafi sh 425Hans-Georg Simon, Molly Ahrens, and Brandon Holtrup50 Vascular and cardiac studies in Drosophila 432Lin Yu, Joseph P. Daniels, and Matthew J. WolfIndex 441