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INFORMATION to USERS the Most Advanced Technology Has Been Used to Photo­ Graph and Reproduce This Manuscript from the Microfilm Master INFORMATION TO USERS The most advanced technology has been used to photo­ graph and reproduce this manuscript from the microfilm master. UMI films the text directly from the original or copy submitted. Thus, some thesis and dissertation copies are in typewriter face, while others may be from any type of computer printer. The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleedthrough, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send UMI a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, charts) are re­ produced by sectioning the original, beginning at the upper left-hand corner and continuing from left to right in equal sections with small overlaps. Each original is also photographed in one exposure and is included in reduced form at the back of the book. These are also available as one exposure on a standard 35mm slide or as a 17" x 23" black and white photographic print for an additional charge. Photographs included in the original manuscript have been reproduced xerographically in this copy. Higher quality 6" x 9" black and white photographic prints are available for any photographs or illustrations appearing in this copy for an additional charge. Contact UMI directly to order. University Microfilms International A Bell & Howell Information Company 300 North Z eeb Road, Ann Arbor, Ml 48106-1346 USA 313/761-4700 800/521-0600 Order Number 9011227 Cell culture models for ueural trauma Murphy, Eric James, Ph.D. The Ohio State University, 1989 Copyright ©1989 by Murphy, Eric James. All rights reserved. UMI 300 N. Zeeb Rd. Ann Arbor, MI 48106 Cell Culture Models for Neural Trauma DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Eric James Murphy, BA ******* The Ohio State University 1989 Dissertation Committee: Approved By Lloyd A. Horrocks Gerald P. Brierley Richard P. Swenson Adviser Department of Physiological Chemistry Copyright by Eric James Murphy 1989 This work is dedicated to my son Cameron and to my wife Cindy, and to my parents Roger and Sandy ii ACKNOWLEDGMENTS In the course of five years there are many people to thank for helping me in my graduate studies. These people include my coworkers Marilyn, Marianne, Kim, Carolyn, Allen, Royal, Paula and Paul. I reserve special thanks for Yutaka "Ruskie" Hirashima and Carl Bates. They have been there over the past year to make the timely completion of this work possible. I also thank Laura Dugan for her instruction in HPLC techniques. I thank Lynda, Barb and Patty for providing large quantities of cell cultures on an almost impossible schedule. I thank Che Maxwell for her help in the first and last year of my studies. I would especially like to thank my wife, Cindy and son Cameron for their patience during the past year. Cindy did a great job preparing the manuscript and worked long hours to get the job done. I thank my parents for their support and for taking care of Cam this summer. I thank my Dad for instilling in me a work ethic that enabled me to continue to work hard in order to complete the task at hand. Of course I have to thank Dr. Dan Funk for his superb surgical work on my knee and my physical therapists Big Dave and Robyn. I also thank my brother, Brian for his support during my injury and recovery as well as my friends Steve Sutherland, Steve Myers, Mark, Shirley, and Veronica. Lastly, I have to thank Dr. Lloyd Horrocks for his support and understanding. He has prepared me well for a career in science and I am sincerely thankful for this preparation. VITA 13 March 1963 Born - Bucyrus, Ohio 1984 BA (Biology and History) Hastings College Hastings, Nebraska 1985-Present Graduate Research Assistant Department of Physiological Chemi stry The Ohio State University Columbus, Ohio PUBLICATIIONS E.J. Murphy, L. Joseph, R. Stephens, and L.A. Horrocks (1986) Phospholipid Composition of Human Endothelial Cultures of Vascular Tissue. Presented at American Society for Neurochemistry Meeting, Montreal, Canada. Trans. Am. Soc. Neurochem. 17: B139. M. Jurkowitz, L. Dugan, E.J. Murphy, M. Waugh, and L.A. Horrocks (1986) Inhibition of Lysoplasmalogenase by 2-acylglycerophospho- ethanolamine. Presented at American Society for Neurochemistry S atellite Symposium on 11 Molecular Mechanisms of Neuronal Responsivity" in Burlington, Vermont. E.J. Murphy, D.K. Anderson, E.D. Means, and L.A. Horrocks (1987) Pressure Induced Trauma in Neuronal Cell Cultures: A Novel Model for Spinal Cord Injury. J. Neurochem., 48: 550A. E.J. Murphy, R. Stephens, M.S. Jurkowitz, and L.A. Horrocks (1987) Plasmalogen Quantitation in Cell Culture Lipids by Acid Hydrolysis Followed by High Pressure Liquid Chromatography. J. Neurochem., 48: S60B. E.J. Murphy, D.K. Anderson, E.D. Means, and L.A. Horrocks (1988) Pressure Induced Trauma in Neuronal Cell Cultures. Presented at American Society for Neurochemistry Meeting, New Orleans, Louisiana, Trans. Am. Soc. Neurochem., 19: BUB. v E.J. Murphy, E. Roberts, and L.A. Horrocks (1989) Aluminum Silicate Toxicity in Cell Cultures. American Society for Neurochemistry Meeting, Chicago, Illin o is. Trans. Am. Soc. Neurochem., 20: B221. E.J. Murphy, D.K. Anderson, E.D. Means, and L.A. Horrocks (1989) Pressure Induced Trauma in ROC-1 Oligodendroglial Cell Cultures. American Society for Neurochemistry Meeting, Chicago, Illin o is. Trans. Am. Soc. Neurochem., 20: A141. E.J. Murphy and L.A. Horrocks (1989) Cell Culture Model for Mechanical Trauma Induced by Pressure. 7th Annual Neurotrauma Meeting, Phoenix, Arizona. E.J. Murphy and L.A. Horrocks (1989) Pressure Induced Trauma in Cell Cultures. Society for Neuroscience, Phoenix, Arizona. Y. Hirashima, E.J. Murphy, and L.A. Horrocks (1989) Purification of Plasmalogens with Rhizopus delemar Lipase and Naja naja naja Phospholipase Ap. American Society for Neurochemistry, March 5-10, 1989, Chicago, Illin o is. Trans. Am. Soc. Neurochem., 20: A265. M. Jurkowitz-Alexander, H. Ebata, J.S. Mills, E.J. Murphy, and L.A. Horrocks (1989) Solubilization, Purification, and Characterization of Lysoplasmalogen alkenylhydrolase (lysoplasmalogenase) from rat liver microsomes. Biochim. Biophys. Acta, 1002: 203-212. E.J. Murphy and L.A. Horrocks (1989) Mechanisms of Action of CDPcholine and CDPethanolamine on Fatty Acid Release during Ischemia of Brain. In Lipid Mediators in Ischemic Brain Damage and Experimental Epilepsy, N.G. Bazan (Ed.). S. Karger AG, Basel, Switzerland. TABLE OF CONTENTS page DEDICATION................................................................................................... ii ACKNOWLEDGMENTS.................................................................. iii VITA............................................................................................................ v LIST OF TABLES.......................................................................................... ix LIST OF FIGURES...................................................................................... xi LIST OF PLATES........................................................................................ xv CHAPTER I. INTRODUCTION 1 II. COMPOSITION Introduction .............................................................................. 7 Materials and Methods ........................................................... 12 Results........................................................................................ 30 Discussion .................................................................................. 39 III. ALUMINUM SILICATES Introduction .............................................................................. 48 Materials and Methods ........................................................... 59 Results........................................................................................ 62 Discussion .................................................................................. 154 IV. PRESSURE Introduction .............................................................................. 168 Materials and Methods ........................................................... 180 R esults...................................................................................... 182 Discussion .................................................................................. 231 v ii CHAPTER V. CONCLUSION.................................................................................. 241 APPENDICIES A.......................................................................................................... 244 B.......................................................................................................... 251 BIBLIOGRAPHY............................................................................................ 270 v i i i LIST OF TABLES Table page 1. Long phospholipid separation ................................................... 21 2. Short phospholipid separation ................................................ 25 3. Phospholipid composition of cell cultures ........................ 32 4. Plasmalogen content of cell cultures .................................. 33 5. FA composition of the major phospholipids in primary neurons c e lls ........................................................
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