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University Microfilms International A Bell & Howell Information Company 300 North Zeeb Road, Ann Arbor, Ml 48106-1346 USA 313/761-4700 800/521-0600 Order Number 8913623 Analyses of the response of dorsal root afferents following dorsal funiculus lesions and an examination of the sacral parasympathetic nucleus in amphibians Campbell, H. Lee, Ph.D. The Ohio State University, 1989 UMI 300 N. Zceb Rd. Ann Arbor, MI 48106 ANALYSES OF THE RESPONSE OF DORSAL ROOT AFFERENTS FOLLOWING DORSAL FUNICULUS LESIONS AND AN EXAMINATION OF THE SACRAL PARASYMPATHETIC NUCLEUS IN AMPHIBIANS DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of the Ohio State University By H. LEE CAMPBELL, B.S. * * * * * The Ohio State University 1989 Dissertation Committee: Approved By Jacqueline C. Bresnahan, Ph.I Michael S. Beattie, Ph.D. Adviser James S. King, Ph.D. Department of Anatomy George F. Martin, Ph.D. For the God who exists and is not silent ACKNOWLEDGEMENTS I would like to thank Dr. Jacqueline C. Bresnahan and Dr. Michael S. Beattie for sharing their time, knowledge, creativity and good humor with me. I am entirely in the debt of Tina Van Meter, David Norris, Dr. Gail Leedy and John Komon each for their own exceptional technical assistance and each for their friendship. The faculty of the Department of Anatomy has proved to be of great help and encouragement, especially Dr. James S. King and Dr. George F. Martin. The loyal companionship of Tim Lipovsky has provided much in the way of acceptance, solace and motivation. My parents have proved themselves without equal in their commitment to my well being and unshakable confidence. I am delighted to be called their son. Finally, I would like to thank Debby for her patience, encouragement, inclination toward laughter and unselfish love. 1 1 1 VITA Date of Birth...... July 8, 1954 Education........... B.S. in Biology, Oakland University, 1981 Ph.D. in Anatomy, The Ohio State University, 1988 Committees.......... Graduate Committee, Dept, of Anatomy, 1987-1988 Council of Graduate Students, The Ohio State University, 1983-1984 Teaching Experience.. Gross Anatomy for Physical Therapists and Occupational Therapists, 1982-1983 Neuroanatomy for Medical Students, 1983, 1988 Histology for Medical Students, 1987-1988 PUBLICATIONS Campbell, H.L., F.J. Liuzzi, M.S. Beattie, and J.C. Bresnahan (1982) Large circumferential cells of the developing Rana catesbeiana spinal cord are labelled after HRP application to lateral funiculus. Neuroscience Abstracts, 23 5.5, 821. Campbell, H.L., M.S. Beattie, J.C. Bresnahan (1984) The response of dorsal root afferent fibers to dorsal funiculus lesions in developing rana catesbeiana tadpoles. Neuroscience Abstracts, 298.7, 1024. Beattie, M.S., J.C. Bresnahan, H.L. Campbell, K.M. Conway, F.J. Liuzzi, and G. Lopate (1985) Studies of iv growth and regeneration in developing anuran spinal cord. Paper presented: Neural Development, Plasticity & Regeneration Conference. Columbus, Ohio April 19-21. Campbell, H.L., M.S. Beattie, J.C. Bresnahan (1987) Circumferential cells of the developing Rana catesbeiana lumbar spinal cord. Anatomy and Embryology 176:155-163. Campbell, H.L., M.S. Beattie, and J.C. Bresnahan (1988) The response of dorsal root afferents to dorsal funicular lesions in developing and juvenile Xenopus laevis. American Association of Anatomists Abstracts, 23ID, 19a. Campbell, H.L., M.S. Beattie, and J.C. Bresnahan (1988) The response of spared dorsal root afferent collaterals following dorsal funiculus lesions in juvenile xenopus laevis. Soc. Neurosci. Abstracts, 467.7, 1170. FIELDS OF STUDY Major Field: Neuroanatomy Minor Fields: Anatomy, Histology v TABLE OF CONTENTS DEDICATION........................................ ii ACKNOWLEDGEMENTS................................. iii VITA............................................... iv LIST OF FIGURES.................................. viii ABBREVIATIONS..................................... xi INTRODUCTION...................................... 1 CHAPTER PAGE I. A REVIEW OF ANURAN SPINAL CORD NEUROANATOMY.......................... 2 1.1 Introduction................. 2 1.2 Spinal Gray.................. 4 1.3 Distribution of primary afferents..................... 24 1.4 Segmental interconnections... 28 1.5 Ascending spinal systems..... 29 1.6 Descending supraspinal systems....................... 35 1.7 Developmental considerations. 42 1.8 Conclus ion.................... 44 II. REGENERATION AND PLASTICITY IN THE DEVELOPING AND ADULT AMPHIBIAN SPINAL CORD...................................... 46 2.1 Introduction................. 46 2.2 Rana catesbeiana............. 57 2.2.1 Methods................ 59 vi CHAPTER PAGE II. 2.2.2 Results............... 63 2.2.3 Discussion............ 68 2.2.4 Conclusion............ 73 2.3 Xenopus laevis................ 75 2.3.1 Methods............... 76 2.3.2 Results............... 81 2.3.3 Discussion............ 89 2.3.4 Conclusion............ 99 III. OBSERVATIONS ON THE XENOPUS LAEVIS SACRAL SPINAL CORD........................... 100 3.1 Introduction.................. 100 3.2 Sacral parasympathetic nucleus........................... 101 3.2.1 Methods............... 103 3.2.2 Results............... 104 3.2.3 Discussion............ 107 ILLUSTRATIONS........................................... 110 BIBLIOGRAPHY............................................. 165 LIST OF FIGURES PAGE Ranid experimental design........... 111 Illustration of the normal distribution of the ninth DR ........ 113 Blastema cells labeled with tritiated thymidine .............. 115 HRP labeled DRG axons in a blastema implant............................... 117 Illustrations of aberrant axonal traj ectories......................... 119 Micrographs of aberrant axonal trajectories......................... 121 Illustrations of growth cones and retraction bulbs..................... 123 Micrographs of aberrant fiber morphologies......................... 125 Illustration of Xenopus experimental design.................. 127 An illustration of the process used to digitize an image for analysis... 129 Normal distribution of tenth root primary afferents.................... 131 Normal rostrocaudal distribution of tenth root primary afferents........ 133 viii PAGE Micrographs showing distributions of DR afferents in the DTF and VTF.... 135 Illustration of the range of injury sites sizes.......................... 137 Micrographs of a root entry zone near an injury site................. 139 A graph showing the detected area for the DTF and VTF across groups... 141 Graphs showing the optical density of the DTF and VTF across groups.... 143 Graphs showing varicosity number and size distribution across groups.... 145 Figures showing examples of crossing fibers in normal, 3 day, 5 week, and 13 week cases........................ 147 A graph showing the lengths of contralateral axons across groups... 149 Graphs showing contralateral axonal number and number of branch points.. 151 Illustrations showing the normal distribution of dorsal roots 10, 9 8, 7, 6 and also of the rubrospinal and raphespinal systems............. 153 Micrographs showing the distribution of elements at the tenth segment after thoracic HRP application..... 155 Photographs showing the surgical procedure used for labeling sacral spinal cord neurons................. 157 Neuronal elements labeled after HRP application to the tenth spinal nerve................................. 159 ix FIGURE PAGE 62 Illustration showing neurons labeled after HRP application to the tenth nerve................................. 161 63 A stereo-pair showing the distribution of cell bodies after HRP application to the tenth nerve.. 163 X ABBREVIATIONS CF central field CNS central nervous system DC dorsal columns DF dorsal field DRG dorsal root ganglion or 'DR' for dorsal root DTF dorsal terminal field [a.k.a. dorsal neuropil] EM electron microscopy GFAP glial fibrillary acidic protein LCC large circumferential