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https://theses.gla.ac.uk/ Theses Digitisation: https://www.gla.ac.uk/myglasgow/research/enlighten/theses/digitisation/ This is a digitised version of the original print thesis. Copyright and moral rights for this work are retained by the author A copy can be downloaded for personal non-commercial research or study, without prior permission or charge This work cannot be reproduced or quoted extensively from without first obtaining permission in writing from the author The content must not be changed in any way or sold commercially in any format or medium without the formal permission of the author When referring to this work, full bibliographic details including the author, title, awarding institution and date of the thesis must be given Enlighten: Theses https://theses.gla.ac.uk/ [email protected] ULTRASTRUCTURAL AND CYTOCHEMICAL STUDIES OF GONADAL ORGANIZATION, SPERMATOGENESIS AND SPERMATOZOA OF MOLLUSCS By Sheng Tao Hou BSc, MSc Thesis submitted in canditure for the degree of Doctor of Philosophy in the Faculty of Medicine, University of Glasgow. Department of Anatomy University of Glasgow September, 1991 Copyright © , 1991 Sheng Tao Hou ProQuest Number: 11008033 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a com plete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 11008033 Published by ProQuest LLC(2018). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 DEDICATION This thesis is dedicated to my wife Qiubo, with love and respect, for her understanding, full support, encouragement and secretarial assistance. DECLARATION I hereby declare that this thesis is my own composition, except where duly acknowledged, and has not been submitted for consideration of any other degree in this or any other Universities. Sheng Tao Hou ACKNOWLEDGEMENTS It is a pleasure to thank the many people whose assistance has been invaluable throughout this project. It is not possible to adequately thank my supervisor Dr W. L. Maxwell, for his continued interest, guidance, encouragement, constructive criticism and incredible patience throughout the course of my research. He has been always willing to give up his time to discuss the work, to help with collecting specimen, and to read and comment on many drafts both for publication and this thesis. The idea of this work stemmed from his teaching, for which I shall always be grateful. I wish to express my sincere gratitute to Professor R. J. Scothorne, F.R.S.E., for providing facilities for my research in the Department of Anatomy, University of Glasgow. I thank all of the technical staff in the Department for their help during my time in the department. Particularly, I am indebted to Mr J. McGadey, Mr D. Russell, Mr R. Kerr and Mr 0. Reid for their valuable instruction and assistance with SEM and TEM; to Ms M. Hughes and Mrs M. Houston for photographic help and Mrs C. Crossan for artistic assistance. My thanks are also due to Dr L. Tetley (Department of Zoology) for permission to use the freeze-fracture apparatus in the centralised laboratory of the University of Glasgow; Mrs A. Sim, for her excellent technical help with freeze-fracture and glycogen cytochemistry, and Dr D. Jiang (Aeronautics Department, University of Glasgow) for his help with statistical analysis. I am extremely grateful to Professor J. Allen in the University Marine Biological Station (Millport, Isle of Cumbrae, Scotland) for his timingly supply of materials (listed in Section Materials A, B and C) , and Mr C. P. Palmer in the Department of Palaeontology, British Museum (Natural History), London, for species identification (Scaphopoda, Antalis entalis). I especially thank Dr A. P. Payne, for his constant encouragement and strong support; Dr R. A Smith for his encouragement, stimulating discussion and spending time helping me with introductory literature concerning microtubular studies. I am also grateful to Mr G. Gillepie, Mrs L. Peedle and Ms. B. Robinson, for their ever kindly help during my study in the department, and Dr D. Badran for his companionship over the past three years. Special thanks are due to Mrs D. Maxwell, for her hospitality, generosity and encouragement throughout this period, to which I am indebted and shall always remember. Lastly, I owe a considerable debt of gratitude to my wife, parents and parents-in-law, for their love, endless and wholehearted support and encouragement which helped me through to the end. This research project was supported by a University of Glasgow Postgraduate Research Student Scholarship and an Overseas Research Student Award. I was extremely honoured to receive these awards for which I am most grateful. CONTENTS PART Is TEXT Page No. TITLE PAGE DEDICATION DECLARATION ACKNOWLEDGEMENTS ABSTRACT ........................................... I-VI GENERAL INTRODUCTION ....................................1 (A). Primitive Type of Spermatozoon And Its Spermatogenesis .....................6 (B). Modified Type of Spermatozoon And Its Spermatogenesis ....................11 (C). Organisation of Testis and Ovotestis ..................................13 MATERIALS AND METHODS...................................18 Materials .......................................... 19 (A) . Scaphopoda ................................ 19 (B) . Decapoda .................................. 19 (C). Opisthobranchia ............................19 (D) . Pulmonata ................................. 2 0 Methods.............................................2 0 (A). Light Microscopy ...........................20 (B). Transmission Electron Microscopy (TEM) ......................................21 a). Fixatives for marine molluscan spermatogenic cells ....................22 b). Fixatives for terrestial molluscan spermatogenic cells ....................23 c) . Hypertonic fixative ....................24 Page No. d). Tracers and fixatives for blood-testis barrier studies ......... 25 e). Fixation of the microtubular manchette .............................. 2 7 f). Araldite embedding ..................... 28 g). Specimen orientation and sectioning .... 30 h). Section staining ....................... 32 1) Routine UA-LC Staining ...............32 2) Cytochemical staining for glycogen ...32 (C) . Freeze-fracture ........................... 33 (D). Scanning Electron Microscopy (SEM).......................................35 CHAPTER I Spermatozoon and Spermatogensis in Antalis entalis (Scaphopoda) ............. 37 Introduction ...................................... 3 8 Results ........................................... 4 3 (A) . Testis .................................... 43 (B). Structure of the Spermatozoon .............4 6 (C) . Early Spermatogenesis .................. 49 (D) . Spermiogenesis ............................ 49 (E). Acrosome and MidpieceFormation ............ 51 Discussion ........................................ 53 (A). Comparative Spermatozoan Structure ..................................53 (B) . Spermatogenesis ........................... 57 CHAPTER II Spermiogenesis of Rossia macrosoma (Decapoda) ............................................ 63 Introduction....................................... 64 Results............................................ 68 Page No. (A). Light and Scanning Electron Microscopical Observations .................68 (B). Ultrastructure of Spermiogenesis ..........71 (1). Nuclear morphogenesis .................71 (2). Proacrosome and acrosome formation .............................72 (3). Mitochondrial midpiece and centriole .........................75 (C) . Ultrastructure of the Spermatozoon ........76 (1). Spermatozoon ..........................76 (2). Glycogen distribution .................78 Discussion .........................................80 (A). Spermatozoon and Spermiogenesis ........... 80 (B). Glycogen Deposition ....................... 86 CHAPTER III Spermatogenesis of Archidoris tuberculata. Tritonia plebeia and Aplvsia faciata (Opisthobranchia) ..........................90 Introduction .......................................91 Results ............................................98 (A). Organization of the Ovotestis and Spermatozoon ...........................98 (B). Early Spermatogenesis .................... 103 (C). Spermiogenesis ........................... 104 (1). Nuclear morphogenesis ................104 (2). Acrosome differentiation .............108 (3). Midpiece and tail formation ......... 109 Discussion ........................................ 113 (A). The Organization of Ovotestis ............ 113 (B). Comparative Spermatogenesis .............. 114 (1). Nutritive cell .......................114 Page No. (2). Golgi complex and acrosome formation ...................115 (3). Nuclear morphogenesis ...............120 (4). Pseudo-paracrystalline materials and midpiece formation ..............122 (5). Glycogen distribution ...............126 (6). Centriolar adjunct ..................127 (7). Systematic considerations .......... 129 CHAPTER IV Intercellular Junctions in the Ovotestis of Arion hortensis (Pulmonata) .............131 Introduction ......................................132 Results ..........................................