Avraham Mayevsky Mitochondrial Function In Vivo Evaluated by NADH Fluorescence Mitochondrial Function In Vivo Evaluated by NADH Fluorescence Avraham Mayevsky Mitochondrial Function In Vivo Evaluated by NADH Fluorescence Avraham Mayevsky Faculty of Life Sciences Bar-Ilan University Ramat-Gan , Israel ISBN 978-3-319-16681-0 ISBN 978-3-319-16682-7 (eBook) DOI 10.1007/978-3-319-16682-7 Library of Congress Control Number: 2015938388 Springer Cham Heidelberg New York Dordrecht London © Springer International Publishing Switzerland 2015 T his work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifi cally the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfi lms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. T he use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specifi c statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. The publisher, the authors and the editors are safe to assume that the advice and information in this book are believed to be true and accurate at the date of publication. Neither the publisher nor the authors or the editors give a warranty, express or implied, with respect to the material contained herein or for any errors or omissions that may have been made. Printed on acid-free paper Springer International Publishing AG Switzerland is part of Springer Science+Business Media (www.springer.com) This book is dedicated to my dearest wife, Zipora, for her vision, patience, and support for more than fi ve decades. Without her encouragement, my academic achievements and writing this book would have been impossible to accomplish. Pref ace This book is not a biography of my life: rather, it tells the story of my research activities since my post-doctorate training with Prof. Britton Chance at the University of Pennsylvania, Philadelphia. In this book, I am integrating the knowledge of all scientists, including myself and my research group, who investigated the function of the intracellular organelle—the mito- chondrion—under in vivo conditions. N ormal mitochondrial function is an essential factor in all tissues and organs in the body. Mitochondrial activity provides all cells in the body with the energy source (ATP) to enable normal functions. Mitochondrial dysfunction creates a major pathological state in various organs because of the decrease in this energy source (ATP). Much pathology in the human body is related to dysfunction of the mitochondria and decrease in ATP production. One of the options for evaluating mitochondrial function in vivo is to measure the autofl uo- rescence of the NADH molecule bound to the enzymatic system in the respiratory chain embedded in the mitochondria. This approach to monitor the mitochondria in vivo was started in the early 1950s by the late Prof. Britton Chance from the University of Pennsylvania, Philadelphia. More than 800 papers were published in this research area by various investiga- tors around the world. Most of the studies involved experimental animals, using various protocols, mimicking the pathological state in patients. In the past 20 years, the same methodology has been applied to monitor patients under various pathological states. During the years, I realized that many young graduate students or other scientists who try to enter this specifi c fi eld of research have diffi culties in fi nding the appropriate source of information to perform a well-controlled study using NADH fl uorescence monitoring in vivo. For this reason, I decided to integrate all the relevant information in a book that will be avail- able to all newcomers to this specifi c fi eld of research, that of mitochondrial function in vivo. This is the fi rst time that this subject has been described in detail, citing almost all the papers published in this fi eld. The fi rst two chapters are a general introduction to the signifi cance of mitochondrial func- tion in health and disease, as well as its part in tissue energy metabolism. Chapter 3 contains a historical overview of NADH monitoring ranging from isolated mitochondria in vitro to intact organs in vivo. As I found many years ago, all optical techniques, and especially monitoring of NADH fl uorescence, when applied to in vivo conditions are very sensitive to the appropriate performance of the experimental procedures. Therefore, all the important technological aspects of NADH monitoring are presented in great detail in Chap. 4 . After the development of the fi ber-optic-based fl uorometer, I realized as a physiologist that monitoring of NADH alone would not provide enough information to understand the patho- physiology of organs in vivo. Therefore, I developed the multiparametric monitoring approach enabling measurement, in addition to NADH fl uorescence, of other physiological parameters at the tissue level in vivo. Chapter 5 is devoted to this subject. In Chap. 6 , the ability to measure NADH as well as other parameters from more than one site simultaneously is presented. In Chapter 7 , which is the largest chapter in the book, I accumulated all the information regarding the responses of NADH as well as other parameters to various perturbations induced in various organs of experimental animals. Most of the results in NADH monitoring were vii viii Preface accumulated in brain studies; therefore, I selected most of the figures from those studies. In each subject, I started with a few representative results published by other researchers, followed by my own studies. It is important to note that in all studies describing the responses of NADH to various perturbations, a typical analog tracing of the responses is presented in parallel to the quantitative analysis of the results and its statistical analysis. I decided to present mainly the real-time results that appeared on paper recorders or were accumulated by comput- erized systems. T o help the reader to fi nd relevant results to the new studies, I prepared appropriate tables, presented in Chap. 8 , that classifi ed all the data accumulated according to the animal or organ selected as well as according to the perturbations used. I n Chap. 9, the use of NADH monitoring in various organs of the human body are presented in historical perspectives and with the up-to-date technology developed during the past 10 to 20 years. In the last chapter, Chap. 1 0 , a short discussion and future perspectives are presented. Ramat-Gan, Israel Avraham Mayevsky Acknowledgments I dedicate this book to my four children, Amotz, Sarit, Hagit, and Shalev, who understood my devotion to an unusual career and enabled me to fulfi ll all my obligations and academic activities. My deep appreciation is given to my parents, Shabtai and Rachel Mayevsky, for their devo- tion and support throughout my life. My appreciation is given to the late Prof. Britton Chance, an outstanding scientist, teacher, and colleague, for almost four decades of fruitful collaboration. During the 40 years of research activities at Bar Ilan University I trained many M.Sc. and Ph.D. students. I want to thank all of them for their confi dence and their efforts during the experimental work as well as in preparation and publication of the manuscripts. Special appreciation is given to a few of my graduate students who joined my research team after their graduation. Their assistance in training new undergraduate and graduate students is appreciated very much. Nili Zarchin, M.Sc., joined my laboratory in the early 1980s. Judith Sonn, Ph.D., started her activities in my laboratory in the early 1990s. Efrat Barbiro-Michaeli, Ph.D., started her activities in the mid-1990 and collaborated with me until my nomination as Professor Emeritus at Bar Ilan University a few years ago. During the past few years, she took part in the preparation for writing this book, and her help is appreciated. P rof. Gennady Rogatsky immigrated to Israel in the early 1990s and was active in my labo- ratory until his retirement. He was involved in research activities and helped me to establish the literature classifi cation for this book. My appreciation is given to his contributions. Special thanks and deep appreciation are given to Eli Zimerman, from Bar Ian University, for his long-term support in the development and maintenance of various instruments in my laboratory that enabled us to perform our studies. I want to thank Mrs. Avrille Goldreich, from the Faculty of Life Sciences at Bar Ian University, for her many years of help and support in the preparation of my manuscripts and especially this book. F inally yet importantly, I express my sincere gratitude to Yossi Mor, who introduced me to Jack Lahav, Co-Chairmen of Israel Venture Partners. This team is supporting me to translate the science described in this book into a revolutionary medical device that will improve the health of humankind. ix