Resource Book for the Design of Animal Exercise Protocols
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American Physiological Society Resource Book for the Design of Animal Exercise Protocols February 2006 i ii AMERICAN PHYSIOLOGICAL SOCIETY COMMITTEE TO DEVELOP AN APS RESOURCE BOOK FOR THE DESIGN OF ANIMAL EXERCISE PROTOCOLS Kevin C. Kregel, PhD, (Chair), The University of Iowa, Iowa City, Iowa; (Chair), Animal Care and Experimentation Committee of the American Physiological Society David L. Allen, PhD, University of Colorado-Boulder, Boulder, Colorado Frank W. Booth, PhD, University of Missouri, Columbia, Missouri Monika R. Fleshner, PhD, University of Colorado-Boulder, Boulder, Colorado Erik J. Henriksen, PhD, University of Arizona, Tucson, Arizona Timothy I. Musch, PhD, Kansas State University, Manhattan, Kansas Donal S. O’ Leary, PhD, Wayne State University, Detroit, Michigan Christine M. Parks, DVM, PhD, University of Wisconsin, Madison, Wisconsin David C. Poole, PhD, DSc, Kansas State University, Manhattan, Kansas Alice W. Ra’anan, American Physiological Society, Bethesda, Maryland Don D. Sheriff, PhD, The University of Iowa, Iowa City, Iowa Michael S. Sturek, PhD, Indiana University School of Medicine, Indianapolis, Indiana Linda A. Toth, DVM, PhD, Southern Illinois University, Springfield, Illinois iii iv PREFACE This resource book was developed to provide information on generally accepted practices for the design and implementation of exercise research protocols involving animals. It is intended to address experimental paradigms for the most commonly used species and models in the context of U.S. national regulatory requirements for animal welfare and to be used in the context of the Animal Welfare Act, the Public Health Service Policy on Humane Care and Use of Laboratory Animals, or other relevant regulations and guidelines. This project grew out of recommendations submitted by the Environmental and Exercise Physiology (EEP) Section of the American Physiological Society (APS) in 2001 to the APS Council. On the basis of a review of published studies involving exercise in various animal species, members of the EEP Section suggested the development of ethical guidelines to be applied in determining whether a given study should be published in APS journals. It has been a long-standing requirement of the APS “Guiding Principles for the Care and Use of Animals” that research studies published in APS journals must minimize the pain and discomfort of animals, but the Society has not offered specific guidance. APS “Ethical Policies and Procedures” are contained in the “Information for Authors” section of the Society’s journal publication instructions and available online at http:// www.the-aps.org/publications/APSEthicalPolicies.pdf. The APS “Guiding Principles for the Care and Use of Animals” can be found online at http:// www.the-aps.org/pa/humane/pa_aps_guiding.htm. The APS Council asked the APS Animal Care and Experimentation (ACE) Committee to examine the EEP recommendations in the broader context of existing regulations and guidelines. In 2002, ACE Committee Chair John N. Stallone asked Kevin Kregel as a member of both the EEP Section and the ACE Committee to organize this project. On December 3, 2002, a planning committee met at the Bethesda, MD headquarters of the APS. The group defined the parameters of the project as providing guidance rather than rules, developed a preliminary outline for the document, and identified additional areas of expertise that were needed. The NIH Office of Laboratory Animal Welfare agreed to provide support for the development of a resource concerning generally accepted standards for animal models of exercise research. v During the next several months, a committee to develop a resource for animal exercise protocols was formed, drawing upon the expertise of the planning committee and others. A first draft was prepared, with committee members contributing sections consistent with their expertise. The expanded project group held a workshop on June 18, 2003. After reviewing the initial draft, the group revised the structure of the document and made additional writing assignments. The third content development workshop was held March 8, 2004 to review progress, discuss broad themes, and identify topics that required further attention. Kevin Kregel, Linda Toth, Erik Henriksen, and Alice Ra’anan served as editors and participated in an additional small meeting that was held on June 1, 2004 for editorial review of content. Authors were then asked to make revisions, and the document was subsequently reviewed by science writer Deborah Berlyne. A final editorial meeting was held on January 27, 2005. The document then underwent peer review, which was administered by Kenneth Baldwin. A diverse group of reviewers was chosen for both technical expertise and broad perspectives related to animal care and exercise testing issues. Comments obtained from these individuals were evaluated and integrated into the final draft of the document, which was completed during the summer of 2005. vi Acknowledgments The authoring committee thanks the APS Council for sponsoring this project and the NIH Office of Laboratory Animal Welfare for its generous support. The committee acknowledges the contribution of Deborah Berlyne, who provided many helpful comments and suggestions, as well as the secretarial assistance of Joan Seye. Appreciation is also extended to Kenneth Baldwin, PhD, University of California at Irvine, for oversight of the peer review process. The committee acknowledges the contributions of the following external reviewers: Paul S. Cooper, DVM, The University of Iowa; V. Reggie Edgerton, PhD, University of California at Los Angeles; John O. Holloszy, MD, Washington University; M. Harold Laughlin, PhD, University of Missouri; Roland R. Roy, PhD, University of California at Los Angeles; Kem B. Singletary, DVM, MS, The University of Iowa; and Charles M. Tipton, PhD, University of Arizona. They provided many constructive comments and suggestions. Appreciation is also extended to Drs. Edgerton and Roy for their input to the contents of this document. A document of this magnitude requires substantial contributions from experts in many different areas. Appreciation is extended to the committee members who took part in various aspects of this project, from participation in the workshops to the drafting and editing of sections of this document. In addition, a special debt of gratitude is owed to Erik Henriksen, Tim Musch, and Linda Toth, who contributed a great deal of time and effort to all facets of this project. Finally, the committee especially acknowledges the efforts of APS Public Affairs Officer Alice W. Ra’anan, who managed the project from beginning to end and was critical to its successful completion. Kevin C. Kregel, Chair Committee to Develop an APS Resource Book for the Design of Animal Exercise Protocols vii viii TABLE OF CONTENTS CHAPTER 1. INTRODUCTION AND OVERVIEW I. Scope of the Document................................................................... 1 II. Why Study Exercise? ...................................................................... 1 III. Why Use Animals to Study Exercise? ............................................ 2 IV. Animal Research Oversight ............................................................ 2 CHAPTER 2. GENERAL ASPECTS OF ANIMAL CARE AND THE DEVELOPMENT OF ANIMAL USE PROTOCOLS I. Overview ........................................................................................ 7 II. Selecting Animal Models for Exercise Research ........................... 7 III. Study Design Considerations.......................................................... 8 A. Protocol Development .............................................................. 8 B. General Considerations for Exercise Study Design.................. 9 IV. Animal Stress and Humane Study Design .................................... 15 A. Stress and Exercise ................................................................. 15 B. Subject and Stressor Characteristics ....................................... 16 C. Indexes of Acute and Chronic Stress Responses .................... 17 V. Working with Compromised Animals: Animal Models of Disease that Limit Exercise Performance ...... 18 A. Special Considerations for Training Compromised Animals ... 18 B. Disease Models that Limit Exercise Capabilities ................... 19 VI. Impact of Surgery on Exercise...................................................... 20 A. Surgical Procedures on Animals Used in Exercise Studies .... 20 B. Selecting Anesthesia for Animals in Surgery ......................... 20 CHAPTER 3. EXERCISE PROTOCOLS USING RATS AND MICE I. Overview ...................................................................................... 23 II. Exercise Modalities in Rats .......................................................... 23 A. Treadmill Running in Rats...................................................... 23 B. Exercise Wheel Running in Rats ............................................ 30 C. Swimming in Rats .................................................................. 35 ix III. Exercise Modalities in Mice ......................................................... 41 A. Background and Introduction ................................................. 41 B. Treadmill Running in Mice .................................................... 43 C. Exercise Wheel Running in Mice ........................................... 48 D. Swimming in Mice ................................................................. 53 IV. Hindlimb Suspension and Immobilization