Development of Sea Urchins, Ascidians, and Other Invertebrate Deuterostomes: Experimental Approaches

909 Pages·2004·11.4 MB·English

by  Charles A. Ettensohn, Gregory A. Wray, and Gary M. Wessel (Eds.)

Methods in Cell Biology
VOLUME 74
Development of Sea Urchins, Ascidians, and Other Invertebrate
Deuterostomes: Experimental Approaches
Series Editors
Leslie Wilson
Department of Biological Sciences
University of California
Santa Barbara, California
Paul Matsudaira
Whitehead Institute for Biomedical Research
Department of Biology
Massachusetts Institute of Technology
Cambridge, Massachusetts
Methods in Cell Biology
Prepared under the Auspices of the American Society for Cell Biology
VOLUME 74
Development of Sea Urchins, Ascidians, and Other Invertebrate
Deuterostomes: Experimental Approaches
Edited by
Charles A. Ettensohn
Department of Biological Sciences
Carnegie Mellon University
Pittsburgh, Pennsylvania
Gregory A. Wray
Department of Biology
Duke University
Durham, North Carolina
Gary M. Wessel
Department of Molecular and Cell Biology and Biochemistry
Brown University
Providence, Rhode Island
Elsevier Academic Press
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CONTENTS
Contributors xvii
Preface xxiii
Dedication xxv
1. The Invertebrate Deuterostomes: An Introduction to Their Phylogeny,
Reproduction, Development, and Genomics
Charles A. Ettensohn, Gary M. Wessel, and Gregory A. Wray
I. Introduction 1
II. Phylogeny 2
III. Reproduction 5
IV. Development 8
V. Genomics 10
References 11
PART I Procurement, Maintenance & Culture of Oocytes,
Embryos, Larvae and Adults
2. Care and Maintenance of Adult Echinoderms
S. Anne Bo¨ttger, Charles W. Walker, and Tatsuya Unuma
I. Overview 18
II. Introduction 18
III. Adult Echinoderm Models: Their Reproductive Cycles and Gametogenesis 18
IV. Obtaining Adult Echinoderms 24
V. Maintenance of Adult Echinoderms in Land-Based Systems 26
VI. Care and Handling of Adult Echinoderms 28
References 33
v
vi Contents
3. Echinoderm Eggs and Embryos: Procurement and Culture
Kathy R. Foltz, Nikki L. Adams, and Linda L. Runft
I. Introduction 40
II. Method for Sea Urchins (Class Echinoidea) 41
III. Method for Sea Stars (Class Asteroidea) 53
IV. Method for Brittle Stars (Class Ophiuroidea, Order Ophiurida) 60
V. Method for Sea Cucumbers (Class Holothuroidea) 64
VI. Method for Interspecific Crosses 68
VII. Summary 70
References 71
4. Culture of Echinoderm Larvae through Metamorphosis
Gregory A. Wray, Chisato Kitazawa, and Benjamin Miner
I. Introduction 75
II. Materials for Culturing Algae and Larvae 76
III. Establishing Algal Cultures 78
IV. Culturing Echinoderm Larvae 81
V. Metamorphosis and Beyond 84
References 85
5. Obtaining and Handling Echinoderm Oocytes
Gary M. Wessel, Ekaterina Voronina, and Jacqueline M. Brooks
I. Introduction 88
II. Experimental Preparation 88
III. Methods of Oocyte Collection 92
IV. Sea Urchin Oocytes Cultured In Vitro 98
V. Labeling of Oocyte Components 101
VI. Introduction of Experimental Substances into the Oocytes 110
VII. Concluding Remarks/Outlook 113
References 114
6. Procurement and Culture of Ascidian Embryos
Billie J. Swalla
I. Overview 116
II. Ascidian Development and Metamorphosis 116
III. Larval Tissue Specification 126
IV. Experimental Techniques 129
V. Protocols 132
References 139
Contents vii
7. Culture of Adult Ascidians and Ascidian Genetics
Carolyn Hendrickson, Lionel Christiaen, Karine Deschet, Di Jiang, Jean-Ste´phane Joly,
Laurent Legendre, Yuki Nakatani, Jason Tresser, and William C. Smith
I. Overview 143
II. Introduction 144
III. Culturing Ascidians 145
IV. Induced Developmental Mutants and Natural Variants/Mutants 153
V. Linkage Analysis and Mapping Genes in Ascidians 160
References 169
8. Hemichordate Embryos: Procurement, Culture, and Basic Methods
Christopher J. Lowe, Kuni Tagawa, Tom Humphreys, Marc Kirschner,
and John Gerhart
I. Introduction 172
II. Procurement, Spawning, and Culture of S. kowalevskii 173
III. Procurement, Spawning, and Culture of Ptychodera flava 179
IV. Removal of Vitelline Envelope in S. kowalevskii 182
V. Whole-Mount In Situ Hybridization 183
VI. Preparation of Blocking Reagent for P. flava 190
VII. Materials and Reagents 191
References 192
9. Cephalochordate (Amphioxus) Embryos: Procurement, Culture,
and Basic Methods
Linda Z. Holland and Ju-Ka Yu
I. Overview 195
II. Introduction 196
III. Obtaining Gametes of Branchiostoma floridae 200
IV. Raising Embryos 202
V. Manipulating Embryos 206
VI. Amphioxus Resources Available 212
VII. Concluding Remarks 213
References 213
viii Contents
PART II Embryological Approaches
10. Quantitative Microinjection of Oocytes, Eggs, and Embryos
Laurinda A. Jaffe and Mark Terasaki
I. Introduction 219
II. Methods 220
III. Equipment and Supplies (Prices are as of December, 2002) 230
References 242
11. Blastomere Isolation and Transplantation
Hyla Sweet, Shonan Amemiya, Andrew Ransick, Takuya Minokawa, David R. McClay,
Athula Wikramanayake, Ritsu Kuraishi, Masato Kiyomoto, Hiroki Nishida, and Jonathan Henry
I. Introduction 244
II. Preparation of Mouth Pipettes and Needles 245
III. Removal of Fertilization Envelope of Sea Urchin Eggs 248
IV. Isolation and Recombination of Blastomeres in Sea Urchin Embryos 249
V. Blastomere Isolation and Transplantation in Starfish Embryos 263
VI. Blastomere Isolation and Transplantation in Amphioxus 265
VII. Blastomere Isolation and Transplantation in Ascidians 266
VIII. Lineage Tracing, Blastomere Isolation, and Transplantation
in Hemichordates 267
References 270
12. Isolation and Culture of Micromeres and Primary Mesenchyme Cells
Fred H. Wilt and Stephen C. Benson
I. Introduction 273
II. Isolation and Culture of Micromeres 275
III. Isolation and Culture of Primary Mesenchyme Cells 282
References 284
13. Rapid Microinjection of Fertilized Eggs
Melani S. Cheers and Charles A. Ettensohn
I. Introduction 287
II. Equipment 289
III. Experimental Protocols 292
References 310
Contents ix
14. Methods for Embryo Dissociation and Analysis of Cell Adhesion
David R. McClay
I. Introduction 311
II. Gaining Access to the Egg and Embryo 312
III. Cell Adhesion Assays: Nonquantitative and Semiquantitative 317
IV. Quantitative Centrifugation Assay 320
References 328
PART III Cell Biological Approaches
15. Analysis of Sea Urchin Embryo Gene Expression by Immunocytochemistry
Judith M. Venuti, Carmen Pepicelli, and Vera Lynn Flowers
I. Introduction 334
II. Immunocytochemistry on Whole Embryos 335
III. Immunostaining Small Numbers of Embryos 351
IV. Co-localization 359
V. Troubleshooting 364
References 366
16. Light Microscopy of Echinoderm Embryos
Laila Strickland, George von Dassow, Jan Ellenberg, Victoria Foe,
Peter Lenart, and David Burgess
I. Introduction (D. Burgess and L. Strickland) 372
II. Formaldehyde Fixation of Cleavage Stage Sea Urchin Embryos 372
III. Staining and Imaging Fixed Embryos 382
IV. Simultaneous Fixation and Visualization of the Actin and Microtubule
Cytoskeletons (G. von Dassow and V. Foe) 385
V. Observation of Live Embryos (D. Burgess and L. Strickland) 393
VI. 4-D Imaging of Fluorescent Markers in Live Starfish Oocytes
( J. Ellenberg and P. Lenart) 399
References 407
17. TEM and SEM Methods
Bruce J. Crawford and Robert D. Burke
I. Introduction 412
II. Basic Fixation and Preparation 412
III. Embedding Media 421
IV. Preservation of Extracellular Matrix 424