Medaka : Biology, Management, and Experimental Protocols

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Edition: CD
Format: Hardcover
Pub. Date: 2009-06-30
Publisher(s): Wiley-Blackwell
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Summary

Medaka: Biology, Management, and Experimental Protocols, written by experienced researchers and reviewed by international leaders in the medaka field will provide details on how to set up and maintain medaka colonies in animal facilities, how to troubleshoot systems, how to handle the fish when applied to experimental methods, and most importantly it will introduce the researcher to cutting edge research in basic and applied biology using medaka as a model animal. The book will include well-written descriptions of experimental methods and protocols designed to educate the reader how to understand and handle medaka effectively.Medaka: Biology, Management, and Experimental Protocols will serve as the definitive reference on the species providing essential information on medaka biology, genetics, and genomics, practical guidance to maintenance of fish stocks, and valuable experimental protocols all in a single volume. This book will be a must have addition to the library of fish researchers and those using medaka as a model organism within their laboratories.

Author Biography

Masato Kinoshita is an Assistant Professor in the Division of Applied Biosciences at Kyoto University, Japan.

Kenji Murata is an Assistant Research Biochemist in the Department of Animal Science at the University of California-Davis, USA.

Kiyoshi Naruse is an Associate Professor at the National Institute for Basic Biology, Japan.

Minoru Tanaka is an Associate Professor at the National Institute for Basic Biology, Japan.

Table of Contents

Contributors
Preface
History and Features of Medaka
History
Phylogeny
Phylogeny and distribution of medaka and relatives
Genetic diversity of medaka
Advantage of Medaka as a Model Fish
Advantageous features in general
Color mutants
Introduction and history
Body color and chromatophores
Genes mutated in body-color mutants
Future use of body-color mutants
Wild strains
Inbred strains
History for establishing inbred lines
Characteristics of medaka inbred strains
For those who cannot decide which medaka to use
Polymorphic variation among inbred strains
To generate and maintain medaka inbred strains
Variation among strains
Differences from zebrafish
Medaka Management
How to Obtain Medaka
Obtain medaka from researchers who are culturing medaka
Contact the National Bio-Resource Project (medaka) in Japan
Purchase medaka from commercial vendors (aquarium shops
Catching medaka from the wild
Rearing Medaka
Breeding program
Recirculating system (mid-scale system
Aquarium system
Maintenance of recirculating system
Soft water is suitable for medaka breeding
Large-scale breeding
Outline of large-scale water system at JST Kyoto
Water system at JST Kyoto facility
Water condition
Rearing without water circulation (small-scale system
Room condition, racks, and tanks
Water
Daily care
Outdoor breeding
Feeding
Feed for adult fish and larvae
Feeding schedule
Feed
Brine shrimp (Artemia
Dry feed
Paramecium
Other feed types
Diseases
Tail rot disease
Matsukasa disease
Trichodina
Water mold
White spot disease
Water mites
Gyrodactylus
Goods for Medaka
Reproduction of Medaka
Sex Determination
Sex determination in medaka
Sex determination in the genus Oryzias
Spontaneous sex reversals in medaka
Hormonal Control of Gonadal Development
Hypothalamicûpituitaryûgonadal axis
Oocyte growth and maturation
Spermatocyte growth and maturation
Oogenesis and Spermatogenesis
Oogenesis
Spermatogenesis
Egg Envelope (Chorion
Morphology and biochemical characters of the medaka egg envelope
Origin of the egg envelope in medaka fish
Gene structure of egg envelope glycoproteins in medaka
Molecular mechanisms of liver-specific expression of ChoriogeninsÆ Genes
Assembly of the Choriogenins into the egg envelope in the ovary
Egg envelope glycoproteins as the substrates for the hatching enzyme
Conclusion
Necessary Conditions for Spawning
Reproductive Behavior
Mating
Embryo Collection
Embryo collection directly from females
Embryo collection from the bottom of the tank
Embryonic Culture
Cleaning
Incubation
Larval Culture
Generation of Sex-Reversed Medaka
Treatment with androgen to generate XX males
Treatment with estrogen to generate XY females
High-temperature treatment to generate XX males
Interstrain Variation in Reproductive Performance
Strain Preservation and Related Techniques
Shipping
Scheduling
Sorting of eggs or fish
Procedures for packaging
Transgenic medaka
MTA (Material Transfer Agreement
Quarantine and Pasteurization
Materials for pasteurization of eggs/embryos
Procedure (Movie M4-1
Cryopreservation of Medaka Sperm
Overview
The procedure for cryopreservation (Figure 4-2 and Movies M4-2
Materials
Solutions
Procedures
Artificial Insemination Using Frozen Medaka Sperm
Overview
Solutions
Materials
Procedures (Movies M4-3
Infertile mating method for collecting unfertilized eggs
Looking at Adult Medaka
General Morphology
Secondary sexual characters
Body color
Pigment cells (chromatophores
Structures of the chromatophores
Chromatophores in medaka
Chromatophore distribution in medaka
See-through medaka
Anatomy and Histology
Observations of internal organs
Observations of internal organs in the live see-through medaka
Dissection of adult medaka
Horizontal and sagittal sections of juvenile medaka
Nervous system
Adult central nervous system
Adult peripheral nervous system
Endocrine system
Hypothalamoûpituitary system
Pineal organ (epiphysis
Thyroid gland
Heart
Interrenal gland and chromaffin cells
Gonads
Endocrine pancreas (islet of Langerhans
Gastrointestinal tract
Ultimobranchial gland
Corpuscle of Stannius
Urophysis
Thymus
Gonads
Ovary
Testis
Kidney
Pronephros
Mesonephros
Histology of the kidney
How to make sections of a meture ovary for histological analysis
Looking at Medaka Embryos
Development of Various Tissues and Organs
Developmental stages
Stage 0; Unfertilized Egg û Figure 6-1
Stage 1; activated egg stage (3 minutes) û Figure 6-1
Stage 2; blastodisc stage û Figure 6-1
Stage 3; two-cell stage (1 hour 5 minutes) û Figure 6-1
Stage 4; four-cell stage (1 hour 45 minutes) û Figure 6-1
Stage 5; eight-cell stage (2 hours 20 minutes) û Figure 6-1
Stage 6; 16-cell stage (2 hours 55 minutes) û Figure 6-2
Stage 7; 32-cell stage (3 hours 30 minutes) û Figure 6-2
Stage 8; early morula stage (4 hours 5 minutes) û Figure 6-2
Stage 9; late morula stage (5 hours 15 minutes) û Figure 6-2
Stage 10; early blastula stage (6 hours 30 minutes) û Figure 6-2
Stage 11; late blastula stage (8 hours 15 minutes) û Figure 6-2
Stage 12; pre-early gastrula stage (10 hours 20 minutes) û Figure 6-3
Stage 13; early gastrula stage (13 hours) û Figure 6-3
Stage 14; pre-mid-gastrula stage (15 hours) û Figure 6-3
Stage 15; mid-gastrula stage (17 hours 30 minutes) û Figure 6-3
Stage 16; late gastrula stage (21 hours) û Figure 6-3
Stage 17; early neurula stage (1 day 1 hour) û Figure 6-3
Stage 18; late neurula stage (1 day 2 hours) û Figure 6-4
Stage 19; two-somite stage (1 day 3 hours 30 minutes) û Figure 6-4
Stage 20; four-somite stage (1 day 7 hours 30 minutes) û Figure 6-4
Stage 21; six-somite stage (1 day 10 hours) û Figure 6-4
Stage 22; nine-somite stage (1 day 14 hours) û Figure 6-4
Stage 23; 12-somite stage (1 day 17 hours) û Figure 6-4
Stage 24; 16-somite stage (1 day 20 hours) û Figure 6-5
Stage 25; 18û19-somite stage (2 days 2 hours) û Figure 6-5
Stage 26; 22-somite stage (2 days 6 hours) û Figure 6-5
Stage 27; 24-somite stage (2 days 10 hours) û Figure 6-5
Stage 28; 30-somite stage (2 days 16 hours) û Figure 6-5
Stage 29; 34-somite stage (3 days 2 hours) û Figure 6-5
Stage 30; 35-somite stage (3 days 10 hours) û Figure 6-6
Stage 31; gill blood vessel formation stage (3 days 23 hours) û Figure 6-6
Stage 32; somite completion stage (4 days 5 hours) û Figure 6-6
Stage 33; stage at which notochord vacuolization is completed (4 days 10 hours) û Figure 6-6
Stage 34; pectoral fin blood circulation stage (5 days 1 hour) û Figure 6-6
Stage 35; stage at which visceral blood vessels form (5 days 12 hours) û Figure 6-6
Stage 36; heart development stage (6 days) û Figure 6-7
Stage 37; pericardial cavity formation stage (7 days) û Figure 6-7
Stage 38; spleen development stage (8 days) û Figure 6-7
Stage 39; hatching stage (9 days) û Figure 6-7
Stage 40; first larval stage û Figure 6-8
Stage 41; second larval stage û Figure 6-8
Stage 42; third larval stage û Figure 6-8
Stage 43; first juvenile stage û Figure 6-8
Stage 44; second juvenile stage û Figure 6-8
Stage 45 û Figure 6-8
Brain
Gastrula step (stages 13û17
Neurula step (stages 17û18
Neural rod step (stages 19û22
Neural tube step (stages 23û27
Late embryonic brain step (stages 28û34
Larval brain step (stages 35û42
Hatching gland
Origin of fish hatching gland cells
Secretion of hatching enzymes from hatching gland cells
Eye development
Specification of the anterior neural plate
Eye field determination and establishment of retinal identity
Splitting of the retinal anlage into two retinal primordia
Morphogenesis I: evagination of the optic vesicle
Morphogenesis II: formation of the optic cup
Retinal differentiation I: central retina
Retinal differentiation II: CMZ
Retinotectal projection
Branchial arch and jaws
Skeletal development
Muscle development
Vasculature
Vascular anatomy of the developing medaka
Origin of the medaka endothelial lineage
Abbreviations
Acknowledgment
Blood cells (hematopoiesis
Overview
Observation of Embryonic and Adult Blood Cells
Heart
Overview
Heart architecture
Heart morphogenesis
Observation of the developing heart
Kidney
Introduction
Nephrogenesis
Pronephros
Mesonephros
Thymus
Overview
Early development of the thymus
Cortex and medulla
Involution of the thymus
Gut and liver
Bones
Vertebral column
Key words in bone formation
Fins
Introduction
Fin anatomy
Embryonic fin development (from fertilization to stage 39 [hatching stage]
Fin development after hatching (after stage 39
Gene expression during fin development
Gonads
Introduction
PGC specification
Formation of gonadal primordium (Figure 6-60B
Sexual dimorphism in germ cell proliferation (Figure 6-61
Post-hatching period in XX gonads
Post-hatching period in XY gonads
Medaka EGG Envelope and Hatching Enzyme
Overview
Preparation of a hatching enzyme solution from hatching liquid
Simple method for preparing hatching enzyme solution
Solubilization of the egg envelope using hatching enzyme
Easy method for preparation of a small amount of hatching enzyme solution (see DVD for figure
Observation of Embryos (Embedding Embryos
Anesthesia of Embryos using MS-222
Observation of embryos (mounting
Living embryos
Processed Embryos
Whole Mount in situ Hybridization (see Section 6.1.8. for a similar protocol
Fixation and storage
Rehydration, proteinase K Treatment and post-fixation at RT
Hybridization and washing
Immunoreaction and washing antibodies
Staining
Embedding in a Plastic Resin (Technovit 7100
Agarose mounting (Figure 6-68
Dehydration and infiltration (Figure 6-68
Polymerization (Figure 6-68
Pigment cells (Figure 6-69
KupfferÆs vesicle
Transgenesis
Microinjection Technique for Medaka Eggs
Equipment required
Egg holder
Glass needles for microinjection
Injector and manipulator with needle holder
Microscope and light
Other tools and fertilized eggs
Microinjection procedure
Microinjection into nuclei
DNA Microinjection
DNA microinjection for transgenesis and transient expression
DNA construction for transgenesis
Toxicity of DNA
The form of DNA for transgenesis
RNA Microinjection
Importance of 3_-UTR
Gene Knockdown Technology
Morpholinos
gripNAs
Toxicology
Status of Medaka in Toxicology
Fish Culture for Toxicology
Preparation and acclimation of fish
How to expose to chemicals
Standardized Toxicity Testings
International standardization for toxicity tests
Acute Toxicity Test (OECD TG203
Early-life stage toxicity test (OECD TG210
Applied Toxicity Tests for Endocrine Disrupters
Screening assays using medaka
Fish full lifecycle testing (FFLC) using medaka
Sensitive period to estrogen substances in early life stages
Vitellogenin as an Environmental Endocrine Disrupting Chemical Exposure Index
Features of VTG
Vitellogenin measurement
Summary and comments
New Techniques and Other Studies
Application of medaka and olyzias Sp. in seawater. Can medaka survive in seawater?
Bioinformatics
Medaka Genome Project and Genome Sequence Database
Genome database
Polymorphism between the Southern and Northern Japanese populations
How to get BAC/Fosmid clones harboring the target gene
Database for Transcribed Sequences
EST database
Developmental Expression database
Positional Cloning of the Causal Gene in Mutants
Mapping mutants using SSLP and RFLP markers
Creating mapping panel
Identification of the linkage group linked to a mutation using bulk segregation analysis with M markers
Low-resolution mapping
Intermediate-Resolution Mapping
High-Resolution Mapping
In silico chromosome walking
Identification of target gene
Construction of fosmid library
Advanced Techniques
Cell Culture from Medaka Embryo
Flow chart of primary cell culture from medaka embryo
Equipment and materials
Protocol
Notes
In Vitro Spermatogenesis from Primary Spermatocytes
Flow chart of in vitro spermatogenesis from primary spermatocytes
Required equipment and materials for primary culture of primary spermatocytes
Protocol
Single Cell Labeling
Flow chart of single cell labeling
Required equipment and materials
Protocol of single cell labeling
Example of cell labeling and tracing
Imaging of Living Embryos
Flow chart of imaging of living embryos
Fluorescent labeling
Sample preparation
Recording setup
Data analysis
Time-lapse imaging of primordial germ cell migration
Conclusion
Transplantation
Cell transplantation in embryo. Figure 10-11 shows the procedure of cell transplantation in embryo briefly
Scale
Nuclear Transplantation
Equipment and materials
Flow chart of the method
Perspectives
Mutagenesis
Benefits of using medaka
Mutagens that have been used for medaka
Mutagenesis screen using ENU
Tilling (Gene Knockout
Outline of the TILLING method
An example of screening and quality of our library
About SNPs
How to obtain a Medaka TILLING library
Cell Trace Experiment with a Caged Fluorescent Dye During Medaka Gastrulation
Flow chart of cell trace experiment with a caged fluorescent dye
Equipment and materials
Protocol (Figure 10-2
Notes
Guidelines on Using Medaka in Experiments
Internet Websites Related to Medaka Research
Solutions
Inbred Strains, Closed Colonies, and Mutant Strains
Index of Abbreviation
Attributions
Index
Table of Contents provided by Publisher. All Rights Reserved.

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