Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1973 The trs ucture, arrangement and intracellular origin of tubular mastigonemes in Ochromonas minute and Monas sp (Chrysophyceae) Francis Gordon Hill Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Biology Commons, and the Botany Commons Recommended Citation Hill, Francis Gordon, "The trs ucture, arrangement and intracellular origin of tubular mastigonemes in Ochromonas minute and Monas sp (Chrysophyceae) " (1973). Retrospective Theses and Dissertations. 5086. https://lib.dr.iastate.edu/rtd/5086 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. 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Silver prints of "photographs" may be ordered at additional charge by writing the Order Department, giving the catalog number, title, author and specific pages you wish reproduced. 5. PLEASE NOTE: Some pages may have indistinct print. Filmed as received. Xerox University Microfilms 300 North Zeeb Road Ann Arbor, Michigan 48106 74-9119 HILL, Francis Gordon, 1947- THE STRUCTURE, ARRANGEMENT AND INTRACELLULAR ORIGIN OF TUBULAR MASTIGONEMES IN OCHRQMONAS MINUTE AND NONAS SP. (CHRYSOPHYCEAETT^ Iowa State University, Ph.D., 1973 Biology University Microfilms, A XEROX Company, Ann Arbor, Michigan The structure, arrangement and intracellular origin of tubular mastigonemes in Ochromonas minute and Monas sp. (Chrysophyceae) by Francis Gordon Hill A Dissertation Submitted to the Graduate Faculty in Partial Fulfillment of The Requirements for the Degree of DOCTOR OF PHILOSOPHY Major: Cell Biology Approved: Signature was redacted for privacy. In Chcfrge of Major Work Signature was redacted for privacy. Chairman, Advisory Commitfee Cell Biology Program Signature was redacted for privacy. For the Graduate College Iowa State University Ames, Iowa 1973 ii TABLE OF CONTENTS Page INTRODUCTION 1 LITERATURE REVIEW 3 Terminology and Mastigoneme Fine Structure 3 Taxonomic Distribution 5 Differences in Mastigoneme Fine Structure Among Taxa 6 Arrangement of Mastigonemes on the Flagellum 8 Molecular Architecture of Mastigonemes 9 Attachment of Mastigonemes to the Flagella 12 Origin and Assembly of Mastigonemes 13 Release of Mastigonemes to the Flagellum 18 Control of Mastigonemogenesis 20 Mastigoneme Function 20 MATERIALS AND METHODS 24 Culture Conditions 2 4 Deflagellation and Kinetics of Reflagellation and Mastigoneme Transport 25 Light Microscopy 25 Electron Microscopy 26 Sectioned material 26 Whole mount material 27 Microscopy 27 RESULTS 28 General Morphology 2 8 iii Page Mastigoneme Structure, Arrangement and Attachment 29 Mastigoneme Morphogenesis and Transport to the Flagellum 31 Analysis of the Intracellular Location of Pre­ sumptive Mastigonemes During Flagellar Re­ generation and Colchicine Inhibition 34 DISCUSSION 59 Phyletic Relationship of Monas to 0. minute 59 Arrangement of Mastigonemes Relative to their Function 60 Attachment of the Mastigoneme to the Flagellum 61 Intracellular Development, Transport and Release of Mastigonemes 62 Golgi apparatus function 64 Control of mastigoneme arrangement 6 7 Nuclear envelope-perinuclear space function 6 8 Kinetics of Mastigoneme Assembly and Transport 72 CONCLUSION 79 BIBLIOGRAPHY 80 ACKNOWLEDGMENTS 91 1 INTRODUCTION Hair-like flagellar appendages (mastigonemes) were first reported by Loeffler (65) in 1889 in the chrysophycean Monas. Subsequently, Fischer (34) , using the same method, a mordant and staining technique developed for bacterial flagella, described a variety of arrangements of mastigonemes on the flagella of many taxa. The existence of mastigonemes was still in question however because of the harsh technique that was used to visualize them. Deflandre (24, 25) demonstrated mastigonemes in 19 34 using nigrosine negative staining and Vlk (108), in 1938, confirmed the existence of mastigonemes with darkfield microscopy of living organisms. This light microscopy is discussed in detail in several reviews (16, 17, 93, 94). Early electron microscopy conclusively demon­ strated the reality of mastigonemes (17, 94). The purpose of this dissertation is multifold. I will review the current and pertinent literature concerning the structure, taxonomic occurrence, assembly, and function of mastigonemes and describe the fine structure of the masti­ gonemes of two chrysophycean flagellates, Ochromonas minute and Monas sp. This will include the attachment of masti­ gonemes to and arrangement on the flagellum and their intra­ cellular development and transport to the cell surface. Some general features of the ultrastructure of the two organisms 2 will also be described. Observations on the structure, ar­ rangement and attachment of mastigonemes will be discussed in relation to flagellate motility in general. The insights into several basic problems in cell biology including self- assembly, functions of the nuclear envelope, modes of secre­ tion, Golgi apparatus function and dynamic membrane flow within the cell, will also be discussed. 3 LITERATURE REVIEW Terminology and Mastigoneme Fine Structure Bouck's definition of mastigonemes is used throughout this dissertation, that is to say, "any essentially fila­ mentous appendage that is clearly distinct from the flagellar membrane or its amorphous coverings" (12). Mastigonemes can be classified according to their struc­ ture into two major groups, the complex tubular mastigonemes and the fine fibrous mastigonemes. The features common to all tubular mastigonemes are: 1) a solid, tapering basal region 0.15-0.3 pm long which is often diffuse and rather ill-defined, 2) a tubular shaft 0.5-1.5 um long and 8-25 nm in diameter and 3) two to three fine terminal filaments 0.5-1.0 ym long. The major variables in tubular mastigoneme structure among taxa, that may be of some taxonomic value, are the overall length of the mastigoneme, the number of terminal filaments, the presence of a distinct basal region and the presence of filaments projecting laterally from the shaft. Nontubular or fibrous mastigonemes make up a structural­ ly diverse group. In general they are fine, flexible fila­ ments projecting from the flagellum in a disorganized array. The arrangement of fibrous mastigonemes on the flagellum varies from a precise helical array of tufts in some 4 Eugienophyceae (61, 82) to a "wooly" or tomentous coat in some Haptophyceae and Chlorophyceae (12). There is some ques­ tion whether the less structurally defined fibrous masti- gonemes found in some taxa are bonafide flagellar appendages or amorphous flagellar coatings precipitated into filaments during preparation for electron microscopy. In some small marine flagellates with scaly flagella (Prasinophyceae) Manton et al. (69, 74-78), and Parke and Manton (89, 90) have reported flagellar hairs that do not appear to fit into either of the above two categories. Coarse, often curved hairs with diagonal striations were re­ ported interspersed among the scales on the flagella of these organisms in shadowcast preparations. The attachment region was somewhat tapered although no distinct basal region was reported. Bouck (12) has suggested that if this description is confirmed with negative staining a third class of masti- gonemes may be indicated. The subject of this dissertation is the complex tubular type of mastigoneme and the fibrous type will not be con­ sidered in any depth. The term mastigoneme when used below will refer to the tubular type unless otherwise stated. 5 Taxonomic Distribution Mastigonemes have been reported in a wide variety of taxa of flagellated