8214146 Snider, Ray Michael AMPHETAMINE AND METHYLPHENIDATE EFFECTS ON NEUROMUSCULAR TRANSMISSION IN THE RAT PHRENIC NERVE - DIAPHRAGM PREPARATION: MECHANISTIC STUDIES CORRELATING MUSCLE CONTRACTION, BIOCHEMICAL AND ELECTROPHYSIOLOGICAL RESULTS The Ohio State University PH.D. 1982 University Microfilms International 300 N. Zeeb Road. Ann Arbor, M I 48106 PLEASE NOTE: In all cases this material has been filmed in the best possible way from the available copy. Problems encountered with this document have been identified here with a check mark V . 1. Glossy photographs or pages ^ 2. Colored illustrations, paper or print _____ 3. Photographs with dark background. 4. Illustrations are poor copy______ 5. Pages with black marks, not original copy ___ 6. Print shows through as there is text on both sides of page _____ 7. Indistinct, broken or small print on several pages _____ 8. Print exceeds margin requirements _____ 9. Tightly bound copy with print lost in spine _____ 10. Computer printout pages with indistinct print ______ 11. Page(s) ___________lacking when material received, and not available from school or author. 12. Page(s) ___________seem to be missing in numbering only as text follows. 13. Two pages numbered ___________. Text follows. 14. Curling and wrinkled pages______ 15. Other _________________________________________________________________ University Microfilms International AMPHETAMINE AND METHYLPHENIDATE EFFECTS ON NEUROMUSCULAR TRANSMISSION IN THE RAT PHRENIC NERVE-DIAPHRAGM PREPARATION: MECHANISTIC STUDIES CORRELATING MUSCLE CONTRACTION, BIOCHEMICAL AND ELECTROPHYSIOLOGICAL RESULTS DISSERTATION. Presented in P a rtia l Fu lfillm en t o f the Requirements for the Degree Doctor of Philosophy in the Graduate School o f The Ohio State University By Ray Michael Snider; B.A. ***** The Ohio State University 1982 Reading Committee: Approved By Michael C. Gerald Dennis R. F e lle r Norman J. Uretsky Adviser Lane J. Wallace College o f Pharmacy ACKNOWLEDGEMENTS To Mary Ellen and my parents, Robert and Dorothy, whose - loving devotion and encouragement has been a source of strength to me, and to who I owe a great deal. To Dr. Michael C. Gerald with whom I have shared a productive, professional relationship and who has been profoundly instrumental in shaping my career. To the Faculty o f the Division of Pharmacology. I dedicate this work to Dr. Lauren K. Gerbrandt; without his influence on my l i f e I would not be where I am today. VITA December 28, 1948 ......................... Born - Los Angeles, Calif. 1974 .................................................... B.A. in Psychology, C alifornia State University Northridge, Northridge, California 1974-1975 ......................................... Technician in the Psychobiology Research Laboratory, V.A. Hospital, Sepulveda, California 1975-1976 ......................................... Teaching Assistant, Department of Psychology, California State University Northridge, Northridge, C alifornia 1976-1981 ......................................... Research Assistant, College of Pharmacy, The Ohio State U niversity, Columbus, Ohio PUBLICATIONS Gerbrandt, L.K., Weyand, T.G. and Snider, R.M.: Afferent control o f hippocampal theta rhythms. Soc. Neurosci. Abstr. JL_: 548, 1975. Gerald, M.C., Gupta, T.K. and Snider, R.M.: Tolerance to amphetamine- induced impairment o f rotarod performance in rats . Psychopharmacology 61: 317-318, 1979. Snider, R.M. and Gerald, M.C.: Electrophysiological analysis of (+)-amphetamine effects on neuromuscular transmission in the rat. Pharmacologist 22: 165, 1980. Snider, R.M., Meldrum, M.J. and Gerald, M.C.: Effects of methyl- phenidate on neuromuscular transmission and muscle contraction in v it r o . Fed. Proc. 4(h 278, 1981. Gerald, M.C. and Snider, R.M.: Amphetamine effects on muscle contraction in the incannulated rat phrenic nerve-diaphragm preparation. Fed. Proc. 40: 278, 1981. i i i Snider, R.M. and Gerald, M.C.: Amphetamine effects on acetylcholine release and mammalian neuromuscular transmission. L ife Sci. 29: 1661-1668, 1981. Snider, R.M. and Gerald, M.C.: Amphetamine increases acetylcholine release from the phrenic nerve: Evidence fo r a noradrenergic- mediated response. Soc. Neurosci. Abstr. 7_: 795, 1981. Meldrum, M .J ., Snider, R.M. and Gerald, M.C.: Cholinergic- independent effects of amphetamine on mammalian skeletal muscle contractions. Neuropharmacology, in press. Snider, R.M. and Gerald, M.C.: Studies on the mechanism of (+)- amphetamine enhancement o f neuromuscular transmission: Muscle contraction, electrophysiological, and biochemical studies. J. Pharmacol. Exp. Ther., submitted fo r publication, 1981. Gerald, M.C. and Snider, R.M.: -Adrenoceptor enhancement of acetylcholine release from motor nerves and effects on muscle contractions by amphetamine and norepinephrine. Fed. Proc., submitted for publication, 1981. FIELDS OF STUDY Major Field: Pharmacology Neuropharmacology Neurobiology Neuromuscular Junction Pharmacology Professor, Michael C. Gerald TABLE OF CONTENTS Page ACKNOWLEDGEMENTS. .................................................................................. i i VITA.................................................................................................................. i i i LIST OF TABLES............................................................................................. ix LIST OF FIGURES......................................................................................... x ii Chapter I . INTRODUCTION .................................................................................. 1 Overview ...................................................................................... 1 Physiology o f Neuromuscular Transmission and Muscle Contraction .......................................................... 3 Neuromuscular transmission ..................................... 3 ACh synthesis .............................................................. 9 ACh storage .................................................................. 10 ACh release ...................................................... 11 Methodological considerations ............................. 12 Termination of ACh action ..................................... 18 Muscle contraction ..................................................... 19 Pharmacology o f Neuromuscular Transmission ................. 23 Presynaptic Effects .............................................................. 24 Hemicholinium and related compounds that inhibit ACh synthesis ......................................... 24 Agents which in h ib it the release o f ACh . 24 Agents which enhance the release o f ACh . 25 Postsynaptic Effects .............................................................. 27 Competitive antagonists ......................................... 27 Noncompetitive or depolarizing blockers . 30 Agents which enhance postsynpatic effects o f A C h...................................................................... 31 Amphetamine Effects on Physical Performance in Humans .................................................................................. 32 Overview .......................................................................... 32 Laboratory studies of amphetamine's effects on physical performance in humans ................. 33 v Page Amphetamine Effects on Physical Performance in Laboratory Animals.......................................................... 39 Enhancement o f performance ................................. 40 Impairment of performance ..................................... 41 Amphetamine Effects on Isolated Mammalian Skeletal Muscle .................................................................................. 42 Amphetamine Effects on D irectly Stimulated Muscle Contractions ...................................................................... 52 Neuropharmacological Mechanism o f Amphetamine Action.................................................................................. 53 Adrenergic Mechanisms ......................................................... 53 Release o f catecholamines ....................... 54 Inhibition of reuptake ' ............................ 54 In h ib itio n of monoamine oxidase (MAO). 55 Direct adrenoceptor effects ................................. 55 Amphetamine Effects on Cholinergic Systems .... 55 Effects on autonomic ganglia ............................. 55 Effects on CNS preparations ................... 57 Adrenergic-Cholinergic Interactions ............................. 57 Adrenergic effects on neuromuscular transmission .......................................................... 58 Adrenergic effects on autonomic and central nervous system cholinergic preparations . 62 Mechanisms of NE effects on ACh release. 65 Methyl pheni date ...................................................................... 67 Neuropharmacology o f methylphenidate .... 67 Methylphenidate, performance and neuro­ muscular transmission ..................................... 69 STATEMENT OF THE PROBLEM...................................................................... 72 Chapter I I . MATERIALS AND METHODS.............................................................. 76 Animals ...................................................................................... 76 Muscle Contraction Studies ............................................. 76 General procedure ..................................................... 76 Electrophysiological Studies ......................................... 84 Biochemical