Developmental Biology: A Guide for Experimental Study

305 Pages·2001·3.22 MB·English

Welcome to the electronic
Developmental
version of the laboratory
manual. Developmental Biology,
Biology
A Guide
A GUIDE FOR EXPERIMENTAL STUDY for Experimental Study, by
Mary S. Tyler.
T h i r d E d i t i o n
Each chapter of the text
contains a set of laboratory
exercises that corresponds to a
module in the main part of the
CD.
The text chapters are in PDF
format, with hyperlinks to an
extensive glossary. You can
access these chapters through
the Vade Mecum modules or
through the Contents below.
The printed form of this lab
manual is available through the
publisher, Sinauer Associates
(www.sinauer.com)
Mary S. Tyler
Sinauer Associates • Publisher
Sunderland, Massachusetts 01375
t a b l e o f Contents
Preface x
1. Getting Started
Things you will need 1-1
Your laboratory notebook 1-1
A format for your notebook 1-3
Formal laboratory reports 1-3
Anatomy of a scientific paper 1-3
Some hints about writing 1-4
Using the library 1-5
Accompanying materials 1-6
Selected bibliography 1-7
2. Embryological Tools
Microknives 2-1
Microneedles 2-2
Hairloops 2-2
Pipettes 2-3
Embryo spoons 2-4
Instrument tray 2-4
Retooling metal instruments 2-5
Sterile technique 2-6
The workbench 2-6
Dissecting tools 2-6
Glassware 2-7
Fluids 2-7
Rules for the road 2-8
Accompanying materials 2-8
Selected bibliography 2-8
Suppliers 2-8
Glossary 2-10
Pre-lab questions 2-11
Contents ii
3. Using the Compound Microscope
The microscope 3-1
Safety first 3-2
Stage 3-2
Magnifications 3-2
Oculars 3-2
Objectives 3-3
Koehler illumination 3-4
The condenser 3-4
Iris diaphragm 3-5
Other settings when maximum resolution is not desired 3-5
Oil immersion 3-5
Make your $1000 microscope into a $10,000 instrument for pennies 3-6
Color filters 3-6
Polarizing filters 3-7
Dark-field optics 3-8
Measurement under the microscope 3-8
Care and maintenance of your microscope: “Twelve good rules” 3-9
Accompanying materials 3-9
Selected bibliography 3-10
Suppliers 3-10
Glossary 3-11
Pre-lab questions 3-14
4. Cellular Slime Molds
Life cycle 4-1
The vegetative stage 4-1
Aggregation 4-1
Pseudoplasmodium 4-3
Culmination 4-4
Sexual reproduction: A rare event 4-4
Preparing for your laboratory studies 4-4
Culture procedures 4-5
Experiments using Dictyostelium 4-6
Labeling with vital dyes 4-6
Transecting, grafting, and disaggregating pseudoplasmodia 4-7
Disruption of cyclic AMP levels 4-8
Behavioral studies 4-9
Completing your experiments 4-10
Accompanying materials 4-10
Contents iii
Selected bibliography 4-10
Suppliers 4-12
Glossa ry 4-13
Pre-lab questions 4-17
5. Gametogenesis
Meiosis: An outline 5-1
Mammalian spermatogenesis 5-2
Stereology 5-4
Tunica albuginea 5-4
Seminiferous tubules 5-4
Spermiogenesis 5-5
Interstitial cells 5-6
Comparative sperm morphology 5-7
Mammalian oogenesis 5-7
Follicles 5-9
Theca 5-9
Oocyte 5-9
Corpus luteum 5-11
Histological Hitchcocking 5-11
Accompanying materials 5-11
Selected bibliography 5-12
Suppliers 5-12
Glossary 5-13
Pre-lab questions 5-18
6. Echinoid Fertilization and Development
Collecting 6-1
Fertilization 6-3
Instructions for normal fertilization 6-4
Watching fertilization 6-5
Interfering with fertilization 6-5
Parthenogenesis 6-6
Cleavage, gastrulation, and larval stages 6-8
Accompanying materials 6-9
Selected bibliography 6-9
Suppliers 6-11
Glossary 6-12
Pre-lab questions 6-17
Contents iv
7. Sea Urchin Development—Effects of Ultraviolet Radiation
Ultraviolet radiation in Earth’s atmosphere 7-1
Biological effects of UV radiation 7-2
Mechanisms that protect against damage from UV 7-3
Using sea urchins as a model organism for UV studies 7-3
Setting up your experiments 7-4
Special lightbulbs that can be used 7-5
Filters that can be used to exclude UV-A or UV-B 7-5
Filters that block UV-B 7-6
Filters that block both UV-A and UV-B (UV opaque) 7-6
Filters that do not block either UV-A or UV-B (UV transparent) 7-7
Safety first 7-7
Monitoring your cultures 7-7
Keeping up with the issues 7-8
Accompanying materials 7-8
Selected bibliography 7-8
Suppliers 7-10
Glossary 7-11
Pre-lab questions 7-15
8. Development of the Fruit Fly
Life cycle 8-1
Culturing Drosophila melanogaster 8-2
Collecting eggs 8-3
Collecting chamber 8-3
Mating behavior of adult flies 8-3
Observations of the egg 8-4
The chorion 8-4
Dechorionating an egg 8-5
Embryogenesis 8-6
Cleavage 8-6
Gastrulation 8-6
Later development 8-8
Embryonic staging series 8-8
Larval development 8-9
Anatomy of the larva 8-10
External anatomy 8-11
Internal anatomy 8-11
Development of imaginal discs 8-12
Dissection of imaginal discs 8-14
Contents v
Eversion of imaginal discs 8-15
Whole-mount preparations of imaginal discs 8-15
Chromosome squash from salivary glands 8-16
Pupation 8-16
Accompanying materials 8-18
Selected bibliography 8-18
Suppliers 8-19
Glossary 8-20
Pre-lab questions 8-28
9. Early Development of the Chick
The chick egg 9-1
The female chick reproductive tract 9-4
Cleavage 9-6
Gastrulation 9-7
24-Hour chick whole mount 9-9
Accompanying materials 9-12
Selected bibliography 9-12
Suppliers 9-13
Glossary 9-13
Pre-lab questions 9-20
10. 33-Hour Chick Embryo
Central nervous system 10-1
Circulatory system 10-2
33-Hour whole mount 10-2
Folds and tucks: Morphogenesis proceeds 10-2
Ectodermal structures 10-3
A category unto themselves: Color them green 10-4
Endodermal structures 10-5
Mesodermal structures 10-5
33-Hour serial sections 10-8
Experiment with evolution 10-13
Create a puppet 10-13
Accompanying materials 10-13
Selected bibliography 10-14
Suppliers 10-15
Glossary 10-15
Pre-lab questions 10-21
Contents vi
11. The Living Embryo and Making of Whole Mounts
Incubation of eggs 11-1
Preparing whole mounts 11-2
Fixation 11-5
Washing 11-6
Staining 11-6
Dehydration and clearing 11-7
Mounting 11-7
Accompanying materials 11-8
Selected bibliography 11-8
Suppliers 11-8
Glossary 11-9
Pre-lab questions 11-12
12. Histological Techniques
Fixation 12-1
Washing 12-3
Dehydration and clearing 12-3
Special microwaving techniques that save time 12-4
Paraffin infiltration 12-4
Paraffin molds 12-5
Paraffin embedding 12-6
Mounting and trimming paraffin blocks 12-6
Sectioning 12-7
Mounting sections 12-8
Staining 12-10
Using the hematoxylin, eosin, and alcian blue staining series 12-11
Hematoxylin 12-12
Mounting coverslips 12-12
Accompanying materials 12-13
Selected bibliography 12-13
Suppliers 12-14
Glossary 12-14
Pre-lab questions 12-17
13. Planarian Regeneration
Collecting and maintaining planarians 13-1
Observations of normal anatomy and behavior 13-2
Anatomy 13-2
Behavior 13-4
The regeneration process: Patterns and theories of regeneration 13-5
Contents vii
The regeneration process 13-5
Pattern formation and positional information 13-6
Diffusion gradient model 13-6
Polar coordinate model 13-8
Patterns of regeneration 13-9
Experimental procedures 13-10
Planning your experiments 13-10
Procedures 13-12
Record keeping and analysis 13-13
Accompanying materials 13-13
Selected bibliography 13-13
Suppliers 13-15
Glossary 13-16
Pre-lab questions 13-19
14. Amphibian Development
Breeding 14-1
Environmental hazards affecting amphibian development 14-2
Preparing for the field trip 14-3
Learn about your local amphibians first 14-3
Equipment needed and recording of data 14-6
Equipment for pH measurements 14-6
Come dressed for the field 14-7
Know government regulations 14-7
The field trip 14-7
Rules for the road 14-9
Night sounds 14-9
Back at the laboratory 14-11
Species identification 14-11
Normal development 14-11
Environmental effects on amphibian development 14-16
Adopt an egg mass 14-18
Stay in touch 14-18
Accompanying materials 14-18
Selected bibliography 14-19
Suppliers 14-21
Glossary 14-22
Pre-lab questions 14-25
Contents viii
15. Zebrafish Development
A new model organism 15-1
Fertilization 15-2
Cleavage 15-5
Blastula 15-5
Gastrulation 15-7
Organogenesis 15-8
Preparing for your laboratory studies 15-9
Culture procedures 15-10
Tanks and water 15-10
Sexing adult fish 15-10
Feeding 15-10
Obtaining embryos 15-10
Maintaining embryos 15-11
Raising larvae 15-12
Experiments using zebrafish embryos 15-12
Observing normal development 15-12
Removing chorion 15-13
Temperature experiments 15-13
Chemical teratogens 15-14
Lithium 15-14
Vitamin A 15-15
Alcohol 15-17
Endocrine disruptors 15-17
Testing water below the pipe 15-18
Behavioral studies 15-18
Completing your experiments 15-18
Preparation of whole mounts 15-19
Fixation 15-19
Washing 15-20
Staining procedures 15-20
Clearing and mounting 15-20
Accompanying materials 15-21
Selected bibliography 15-21
Suppliers 15-24
Glossary 15-25
Pre-lab questions 15-36
Contents ix
Web Links to Appendices
Web pages for the appendices below can be found at w ww.developmentalbiology.ne t
The hyperlinks below go directly to each of the individual pages.
Appendix 1: List of Recipes
Appendix 2: List of Suppliers and Addresses
Appendix 3: Full Glossary
Appendix 4: Laboratory Safety