Hydra: Research Methods

Front Matter....Pages i-xxxi
Introduction....Pages 1-3
Terminology for Morphology and Cell Types....Pages 5-14
Front Matter....Pages 15-15
Collecting Hydra....Pages 17-18
Identifying Hydra Species....Pages 19-28
Water, Culture Solutions, and Buffers....Pages 29-34
Visual Monitoring of pH in Solutions with Phenol Red....Pages 35-37
Hatching Brine Shrimp Larvae Axenically and/or in a Range of Quantities....Pages 39-46
Determining Growth Rates of Groups of Hydra and Budding Rates of Individual Hydra....Pages 47-52
Culturing Large Numbers of Hydra....Pages 53-62
Turbidimetric and Pipetimetric Measurements of Number of Hydra....Pages 63-65
Culturing Hydra of the Same Species but of Different Sizes....Pages 67-69
Culture of Sexually Differentiated Hydra....Pages 71-77
Preparing Axenic Hydra....Pages 79-83
Front Matter....Pages 85-85
Preparing Hydra for Transmission Electron Microscopy....Pages 87-94
Preparing Hydra for Scanning Electron Microscopy....Pages 95-104
Preparing Hydra for Freeze-Fracture and Freeze-Etching....Pages 105-115
Whole Mounts for Light Microscopy....Pages 117-120
Preparing Histological Sections for Light Microscopy....Pages 121-130
Vital Staining: Fluorescent and Immunofluorescent, and Review of Nonfluorescent Dyes....Pages 131-140
Front Matter....Pages 141-141
Macrophotography....Pages 143-149
Front Matter....Pages 151-151
Dissociating Hydra Tissue into Single Cells by the Maceration Technique....Pages 153-156
Cell Cycle Analysis of Hydra Cells....Pages 157-163
Mitotic Index....Pages 165-168
Measurement of the Numbers of Nematoblasts, Nematocytes, and Nematocysts....Pages 169-182
Marking Epithelial Cells in Living Hydra with India Ink....Pages 183-186
Front Matter....Pages 187-187
Incorporating [ 3 H]Thymidine into Hydra by Microinjection....Pages 189-191
Labeling with Gaseous 14 CO 2 or by Feeding on Radioactive Tissues....Pages 193-196
Fractionating Small Amounts of Radioactive Tissue....Pages 197-203
Rapid Whole-Mount Radioautography....Pages 205-207
Front Matter....Pages 209-209
Isolating Hydra Mutants by Sexual Inbreeding....Pages 211-221
Front Matter....Pages 223-223
Grafting: A Rapid Method for Transplanting Tissue....Pages 225-232
Quantitative Interpretation of Transplantation Phenomena....Pages 233-249
Dissociating Tissues into Cells and the Development of Hydra from Aggregated Cells....Pages 251-259
Culturing Interstitial Stem Cells in Hydra Aggregates....Pages 261-266
Separating Viable Tissue Layers....Pages 267-271
Preparing Ectoderm/Endoderm Chimeras....Pages 273-277
Front Matter....Pages 279-279
Eliminating All Nonepithelial Cells Using Colchicine....Pages 281-286
Culturing Epithelial Hydra....Pages 287-290
Reducing Populations of Interstitial Cells and Nematoblasts with Hydroxyurea....Pages 291-294
Preparing Hydra viridis with Nerve Cells and No Interstitial Cells, or with Neither of These Cell Types....Pages 295-297
Front Matter....Pages 279-279
Eliminating Interstitial Cells with Nitrogen Mustard....Pages 299-302
Altering Cell Population Levels by Gamma Irradiation....Pages 303-304
Reducing Number of Nematocytes in the Tentacles....Pages 305-307
Front Matter....Pages 309-309
Assay and Isolation of Substances Controlling Morphogenesis in Hydra....Pages 311-324
Front Matter....Pages 325-325
Isolating Mesolamellae....Pages 327-329
Isolating Undischarged and Discharged Nematocysts from Acontiate Sea Anemones....Pages 331-333
Dissolving the Nematocyst Capsule Wall and Identifying its Protein Component(s)....Pages 335-339
Purifying an Inhibitor of Succinoxidase Activity from Hydra littoralis ....Pages 341-346
Assays for Activities of Nematocyst Venoms and Their Components....Pages 347-358
Front Matter....Pages 359-359
Special Techniques for Weighing Microgram Quantities of Tissue and Assaying Them for Enzyme Activities....Pages 361-371
Extracting and Characterizing Hydra RNA: Modifications to Allow Extraction of Undegraded Material in the Presence of High Levels of Degradative Enzymes....Pages 373-377
Colorimetric Analysis for Protein of Hydra....Pages 379-381
Determining Respiration and Oxygen Evolution of Green Hydra with the Rank Brothers Oxygen Electrode....Pages 383-387
Front Matter....Pages 389-389
Isolating Endosymbiotic Algae from Hydra viridis ....Pages 391-392
Preparing Aposymbiotic Hydra....Pages 393-397
Introducing Symbiotic Algae into Aposymbiotic Hydra....Pages 399-400
Measuring Number of Algal Symbionts in Hydra viridis ....Pages 401-405
Measuring in Vivo Translocation of Reduced Organic Carbon Compounds from Endosymbiotic Algae to Hydra....Pages 407-409
Spectrophotometric Assay for Maltose....Pages 411-413
Front Matter....Pages 415-415
Experimental Methods for Epizootilogical Research with Hydras....Pages 417-425
Front Matter....Pages 427-427
Recording Electrical Activity....Pages 429-441
Bioassay for, and Characterization of, Activators and Inhibitors of the Feeding Response....Pages 443-451
Back Matter....Pages 453-463