Mechanisms of reduced opioid effectiveness in a rat model of neuropathic pain Item Type text; Dissertation-Reproduction (electronic) Authors Nichols, Michael Lorne, 1967- Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 24/09/2021 16:21:32 Link to Item http://hdl.handle.net/10150/282342 INFORMATION TO USERS This manuscript has been reproduced from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter &ce, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. 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Contact UMI directly to order. UMI A Bell & Howell Infonnation Company 300 North Zeeb Road, Ann Aibor MI 48106-1346 USA 313/761-4700 800/521-0600 Mechanisms of Reduced Opioid Effectiveness in a Rat Model of Neuropathic Pain by Michael Lome Nichols A Dissertation Submitted to the Faculty of the COMMITTEE ON PHARMACOLOGY AND TOXICOLOGY (Graduate) In Partial Fulfillment of the Requirements For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College THE UNIVERSITY OF ARIZONA 1997 UMI Nximber: 9729518 UMI Microform 9729518 Copyright 1997, by UMI Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. UMI 300 North Zeeb Road Ann Arirar, MI 48103 2 THE UNIVERSITY OF ARIZONA ® GRADUATE COLLEGE As members of the Final Examination Committee, we certify that we have read the dissertation prepared by Michael L. Nichols entitled Mechanisms of Reduced Opioid Effectiveness in a Rat Model of Neuropathic Pain and recommend that it be accepted as fulfilling the dissertation requirement for tlie Degree of Doctor of Philosophy OA.KSLCS. 4-- FV^k Porr^a,^h.D. Date ^9-^7 Edward D. , Ph.D. Date Lai, Ph.D. Date r. PhUlilsCMa^./M.D'./,In,/;M.D^, Ph.D. Date H-lih7 D. Palmer, M.D., Ph.D. Date Final approval and acceptance of this dissertation is contingent upon the candidate's submission of the final copy of the dissertation to the Graduate College. I hereby certify that I have read this dissertation prepared under my direction and re^mmend that it be accepted as fulfilling the dissertation requirement. hAiAlt. f ~/0 -^1 Usfeertation Director Date 3 STATEMENT BY AUTHOR This dissertation has been submitted in partial fulfillment of requirements for an advanced degree at The University of Arizona and is deposited in the University Library to be made available to borrowers under mles of the Library. Brief quotations from this dissertation are allowable without special permission, provided that accurate acknowledgment of source is made. Requests for permission for extended quotation from or reproduction of this manuscript in whole or in part may be granted by the head of the major department or the Dean of the Graduate College when in his or her judgment the proposed use of the m ^ * • ^ ' ' hip. In all other instances, however, permission must be 4 ACKNOWLEDGEMENTS I thank Dr. Frank Porreca for giving me the opportunity to be successful when no one else would. Frank has provided me with an excellent setting to develop and excel as a young scientist. Further, Frank has given me both the guidance, motivation and example of a true mentor. I also thank my conunittee members, Drs. Ed French, Josephine Lai. Phil Malan and John Palmer for their time and effort to help me complete my Ph.D. I also thank Dr. Larry Reid everything he has given me along the way. Di (Chuck) Bian and Michael (Rash) Ossipov deserve special recognition. Chuck has worked with me on nearly every project in the laboratory, and his surgical skills have saved me countless hours of frustration. Mike O. has helped me through my confusion with his expertise and guidance. On the intellectual side of things he has given his all to help me understand. I thank them both very much. I have shared many things with Ed (Dr. Dork) Bilsky, including office space, for the greater part of nearly twelve years. Ed has been a great friend who has kept the lab from feeling like a job. I will dearly miss Ed and his antics and I wish him and his lovely wife, Jill, all the best. Other members of the Porreca tribe that deserve recognition are Joe Tumminia, C.J. (Squeegee) Kovelowski, Dr. Todd (Squirrel) Vanderah, Dr. Robert (Bob) Horvath, Sandy (Flower) Wegert, Eddie Mata, Dr. Ken (Vem) Wild and Jason Lashbrook for always lending a hand or an ear to me along the way. I consider all of you my friends. I also thank all of the friends I have made in Arizona including Ray-ray, Steve Stratton, Lungs of a Smoker (Jill, Kent, AI, Mr. Potatohead and EFang), softball teamates and camping crews. Russ (Crusty) Ingersoll is my dear friend, whom I'll miss very much. Lastly, I thank my best friend, Wanda, who has not only been my friend, but has managed to survive living with me for nearly four years. I love you very much, Wanda. 5 DEDICATION This disseration is dedicated to my parents, Robert and Marie Nichols. Their sacrifice gave me the opportunity to begin my academic career. Their support helped to propel me through my bachelor's, master's, and now, my doctoral degree. I would never have been given these opportunities without them. 6 TABLE OF CONTENTS Page LIST OF ILLUSTRATIONS 13 ABSTRACT 20 INTRODUCTION 22 Nociception 23 The neurotransmission of pain 25 Hyperalgesia 27 Opiates and opioids and pain perception 30 Brief history 30 Opioid receptors and endogenous opioids 31 Opioids and the inhibition of pain 36 Endogenous pain control 40 Abnormal pain states: peripheral neuropathy 41 Animal models of peripheral neuropathy 43 Opiates in neuropathic pain 44 Excitatory amino acids and neuropathic pain 46 7 TABLE OF CONTENTS-ConfmueJ Page The non-opioid effects of dynorphin and neuropathic pain 49 Afferent fibers and neuropathic pain 50 Modulation of the effectiveness of mu opiates (morphine) 52 HYPOTHESIS OF THE DISSERTATION 57 Hypothesis 1 57 Hypothesis 2 57 Hypothesis 3 58 METHODS 59 Animals 59 Chemicals 59 Surgical implantation of the intrathecal catheter 60 Drug administration 61 L5/L6 nerve ligation and sham surgery 62 Neonatal capsaicin treatment 63 Behavioral Testing 63 von Frey assay (including normalized von Frey fibers) 63 8 TABLE OF CONTENTS- Continued Page Warm-water tail flick assay 64 Radiant heat foot flick assay 65 Pin prick assay 65 Statistics 65 RESULTS OF PRELIMINARY EXPERIMENTS 67 Baseline testing 67 Testing with normalized von Frey fllaments 68 Effect of anesthetic on the development of tactile allodynia 71 SUMMARY OF PRELIMINARY EXPERIMENTS 77 RESULTS OF STUDIES INVOLVING OPIATES AND OPIOIDS 79 Lack of effect of intrathecal morphine to probing with von Frey monofilaments 79 Reduced antinociceptive effectiveness of morphine in rats with nerve-ligation injury 83 Studies on the effect of intrathecal [D-Ala^, NMePhe^, Gly- oljenkephalin (DAMGO) to probing with von Frey monofilaments. .83 9 TABLE OF CONTENTS- Continued Page Studies on the antiallodynic effect of intrathecal SNC 80 in rats with nerve injury 88 Studies on the antiallodynic effect of intrathecal [D-Ala^, Glu^ldeltorphin in rats with nerve injury 90 Studies on the antiallodynic effect of intrathecal biphalin in rats with nerve injury 93 Studies on the antiallodynic effect of intrathecally administered kappa agonists in rats with nerve injury 97 SUMMARY OF STUDIES INVOLVING OPIATES AND OPIOIDS 100 RESULTS OF EXPERIMENTS INVOLVING NEONATAL CAPSAICIN TREATMENT 106 Effects of capsaicin treatment on nociceptive thresholds 106 Warm-water Tail Flick {48'C) 106 Warm-water Tail Flick (52"C) 107 Radiant Heat Paw Withdrawal 107 Pin Prick 110 10 TABLE OF CONTENTS- Continued Page Von Frey probing 110 Effects of morphine in capsaicin treated rats 114 Morphine in the warm-water tail flick (52" C) 114 Morphine in the von Frey assay 120 Effects of DAMGO in capsaicin treated rats 123 DAM GO in the warm-water tail flick (52" C) 123 DAMGO in the von Frey assay 129 SUMMARY OF EXPERIMENTS INVOLVING NEONATAL CAPSAICIN TREATMENT 134 RESULTS OF EXPERIMENTS INVOLVING EXCITATORY AMINO ACID ANTAGONISTS AND DYNORPHIN A (1-13) ANTISERA 140 Studies on the antiallodynic effects of excitatory amino acid antagonists 140 MK 801 versus tactile allodynia 140 CNQX versus tactile allodynia 142 L-AP3 versus tactile allodynia 142 11 TABLE OF CONTENTS- Continued Page Studies on the effects of morphine in the presence of MK 801 148 Tactile allodynia 148 Warm-water tail flick (55°C) 151 Studies on the effect of morphine following pretreatment with antisera to dynorphin A (1-13) 154 Tactile allodynia 154 Warm-water tail flick 55"C 154 SUMMARY OF EXPERIMENTS INVOLVING EXCITATORY AMINO ACID ANTAGONISTS AND DYNORPHIN A (1-13) ANTISERA 159 RESULTS OF EXPERIMENTS INVOLVING DYNORPHIN A (1-17) 163 SUMMARY OF EXPERIMENTS INVOLVING DYNORPHIN A (1-17)..