Medaka

Name: Medaka | Biology, Management, and Experimental Protocols, Volume 2
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Biology, Management, and Experimental Protocols, Volume 2

Kenji Murata Masato Kinoshita Kiyoshi Naruse Minoru Tanaka Yasuhiro Kamei(Herausgeber*in)

Wiley-Blackwell (Verlag)

1. Auflage

Erschienen am 10. September 2019

368 Seiten

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Inhalt

Cover
Title Page
Copyright Page
Contents
List of Contributors
Preface
Chapter 1 Medaka Management
1.1 Introduction
1.2 Medaka Management for Scientific Research
1.2.1 Outline of medaka life-cycle in the wild
1.2.2 Preparation of normal rearing conditions of medaka in the laboratory and procedures for breeding
1.2.2.1 Breeding system set-up
1.2.2.2 Obtaining medaka
1.2.2.3 Collecting eggs in a laboratory setting
1.2.2.4 Daily care and maintenance of eggs
1.2.2.5 Rearing medaka from the larval stage to adulthood
1.2.2.6 Anesthesia and euthanasia
1.3 Standardized Culture and Growth Curve
1.3.1 Characteristics and selection of strains
1.3.2 Management of medaka eggs and fish
1.3.2.1 Mating
1.3.2.2 Management of embryos
1.3.2.3 Management of embryos before hatching
1.3.2.4 Rearing from the larval stage to adulthood (to induce earlier maturation)
1.3.3 Maintenance of breeding tanks during breeding
1.3.3.1 Judgment of water quality
1.3.3.2 Maintenance of breeding water
1.3.4 Anesthesia
1.3.4.1 Behavior under each anesthesia stage
1.3.4.2 Difference in sensitivity to anesthesia among strains
1.3.4.3 Growth stage specificity in sensitivity to MS-222
1.3.4.4 Eugenol is recommended as an anesthetic reagent
1.3.4.5 Euthanasia
1.3.4.6 Important reminders for euthanasia
References
Chapter 2 Medaka and Oryzias Species as Model Organisms and the Current Status of Medaka Biological Resources
2.1 Introduction
2.2 Common and Unique Futures of Medaka and Related Species as Model Organisms
2.3 Phylogenetic Relationships of Medaka and Related Species
2.3.1 The javanicus species group
2.3.2 The latipes species group
2.3.3 The celebensis species group
2.4 BAC Resources of Species Related to Medaka
2.5 National Bio-Resource Project Medaka (NBRP Medaka)
2.5.1 Support for visiting researchers
Acknowledgments
References
Chapter 3 Looking at Adult Medaka
3.1 General Morphology
3.1.1 Secondary sexual characters
3.1.1.1 Dorsal fin
3.1.1.2 Anal fin
3.1.1.3 Papillar processes
3.1.1.4 Urogenital papillae
3.1.2 Body color
3.1.2.1 Pigment cells (chromatophores)
3.1.2.2 Structures of the chromatophores
3.1.2.3 Chromatophores in medaka
3.1.2.4 Chromatophore distribution in medaka
3.1.2.5 See-through medaka
3.2 Anatomy and Histology
3.2.1 Observations of internal organs
3.2.1.1 Observations of internal organs in the live see-through medaka
3.2.1.2 Dissection of adult medaka
3.2.2 Horizontal and sagittal sections of juvenile medaka
3.2.3 Nervous system
3.2.3.1 Adult central nervous system
3.2.3.2 Adult peripheral nervous system
3.2.4 Endocrine system
3.2.4.1 Hypothalamo-pituitary system
3.2.4.2 Pineal organ (epiphysis)
3.2.4.3 Thyroid gland
3.2.4.4 Heart
3.2.4.5 Interrenal gland and chromaffin cells
3.2.4.6 Gonads
3.2.4.7 Endocrine pancreas (islets of Langerhans)
3.2.4.8 Gastrointestinal tract
3.2.4.9 Ultimobranchial gland
3.2.4.10 Corpuscle of Stannius
3.2.4.11 Urophysis
3.2.4.12 Thymus
3.2.5 Gonads
3.2.5.1 Ovary
3.2.5.2 Testis
3.2.6 Kidney
3.2.6.1 Pronephros
3.2.6.2 Mesonephros
3.2.6.3 Histology of the kidney
Appendix
References
Chapter 4 Looking at Medaka Embryos
4.1 Development of Various Tissues and Organs
4.1.1 Developmental stages
4.1.1.1 Stage 0: unfertilized egg (Figure 4-1)
4.1.1.2 Stage 1: activated egg stage (3 minutes) (Figure 4-1)
4.1.1.3 Stage 2: blastodisc stage (Figure 4-1)
4.1.1.4 Stage 3: two-cell stage (1 hour 5 minutes) (Figure 4-1)
4.1.1.5 Stage 4: four-cell stage (1 hour 45 minutes) (Figure 4-1)
4.1.1.6 Stage 5: eight-cell stage (2 hours 20 minutes) (Figure 4-1)
4.1.1.7 Stage 6: 16-cell stage (2 hours 55 minutes) (Figure 4-2)
4.1.1.8 Stage 7: 32-cell stage (3 hours 30 minutes) (Figure 4-2)
4.1.1.9 Stage 8: early morula stage (4 hours 5 minutes) (Figure 4-2)
4.1.1.10 Stage 9: late morula stage (5 hours 15 minutes) (Figure 4-2)
4.1.1.11 Stage 10: early blastula stage (6 hours 30 minutes) (Figure 4-2)
4.1.1.12 Stage 11: late blastula stage (8 hours 15 minutes) (Figure 4-2)
4.1.1.13 Stage 12: pre-early gastrula stage (10 hours 20 minutes) (Figure 4-3)
4.1.1.14 Stage 13: early gastrula stage (13 hours) (Figure 4-3)
4.1.1.15 Stage 14: pre-mid-gastrula stage (15 hours) (Figure 4-3)
4.1.1.16 Stage 15: mid-gastrula stage (17 hours 30 minutes) (Figure 4-3)
4.1.1.17 Stage 16: late gastrula stage (21 hours) (Figure 4-3)
4.1.1.18 Stage 17: early neurula stage (1 day 1 hour) (Figure 4-3)
4.1.1.19 Stage 18: late neurula stage (1 day 2 hours) (Figure 4-4)
4.1.1.20 Stage 19: two-somite stage (1 day 3 hours 30 minutes) (Figure 4-4)
4.1.1.21 Stage 20: four-somite stage (1 day 7 hours 30 minutes) (Figure 4-4)
4.1.1.22 Stage 21: six-somite stage (1 day 10 hours) (Figure 4-4)
4.1.1.23 Stage 22: nine-somite stage (1 day 14 hours) (Figure 4-4)
4.1.1.24 Stage 23: 12-somite stage (1 day 17 hours) (Figure 4-4)
4.1.1.25 Stage 24: 16-somite stage (1 day 20 hours) (Figure 4-5)
4.1.1.26 Stage 25: 18-19-somite stage (2 days 2 hours) (Figure 4-5)
4.1.1.27 Stage 26: 22-somite stage (2 days 6 hours) (Figure 4-5)
4.1.1.28 Stage 27: 24-somite stage (2 days 10 hours) (Figure 4-5)
4.1.1.29 Stage 28: 30-somite stage (2 days 16 hours) (Figure 4-5)
4.1.1.30 Stage 29: 34-somite stage (3 days 2 hours) (Figure 4-5)
4.1.1.31 Stage 30: 35-somite stage (3 days 10 hours) (Figure 4-6)
4.1.1.32 Stage 31: gill blood vessel formation stage (3 days 23 hours) (Figure 4-6)
4.1.1.33 Stage 32: somite completion stage (4 days 5 hours) (Figure 4-6)
4.1.1.34 Stage 33: stage at which notochord vacuolization is completed (4 days 10 hours) (Figure 4-6)
4.1.1.35 Stage 34: pectoral fin blood circulation stage (5 days 1 hour) (Figure 4-6)
4.1.1.36 Stage 35: stage at which visceral blood vessels form (5 days 12 hours) (Figure 4-6)
4.1.1.37 Stage 36: heart development stage (6 days) (Figure 4-7)
4.1.1.38 Stage 37: pericardial cavity formation stage (7 days) (Figure 4-7)
4.1.1.39 Stage 38: spleen development stage (8 days) (Figure 4-7)
4.1.1.40 Stage 39: hatching stage (9 days) (Figure 4-7)
4.1.1.41 Stage 40: first larval stage (Figure 4-8)
4.1.1.42 Stage 41: second larval stage (Figure 4-8)
4.1.1.43 Stage 42: third larval stage (Figure 4-8)
4.1.1.44 Stage 43: first juvenile stage (Figure 4-8)
4.1.1.45 Stage 44: second juvenile stage (Figure 4-8)
4.1.1.46 Stage 45 (Figure 4-8)
4.1.2 Brain
4.1.2.1 Gastrula step (stages 13-17)
4.1.2.2 Neurula step (stages 17-18)
4.1.2.3 Neural rod step (stages 19-22)
4.1.2.4 Neural tube step (stages 23-27)
4.1.2.5 Late embryonic brain step (stages 28-34)
4.1.2.6 Larval brain step (stages 35-42)
4.1.3 Hatching gland
4.1.3.1 Origin of fish hatching gland cells
4.1.3.2 Secretion of hatching enzymes from hatching gland cells
4.1.4 Eye development
4.1.4.1 Specification of the anterior neural plate
4.1.4.2 Eye field determination and establishment of retinal identity
4.1.4.3 Splitting of the retinal anlage into two retinal primordia
4.1.4.4 Morphogenesis I: evagination of the optic vesicle
4.1.4.5 Morphogenesis II: formation of the optic cup
4.1.4.6 Retinal differentiation I: central retina
4.1.4.7 Retinal differentiation II: Ciliary Marginal Zone
4.1.4.8 Retinotectal projection
4.1.5 Branchial arch and jaws
4.1.5.1 Skeletal development
4.1.5.2 Muscle development
4.1.6 Vasculature
4.1.6.1 Vascular anatomy of the developing medaka
4.1.6.2 Origin of the medaka endothelial lineage
4.1.6.3 Abbreviations
4.1.6.4 Acknowledgment
4.1.7 Blood cells (hematopoiesis)
4.1.7.1 Overview
4.1.7.2 Observation of embryonic and adult blood cells
4.1.8 Heart
4.1.8.1 Overview
4.1.8.2 Heart architecture
4.1.8.3 Heart morphogenesis
4.1.8.4 Observation of the developing heart
4.1.9 Kidney
4.1.9.1 Overview
4.1.9.2 Nephrogenesis
4.1.9.3 Pronephros
4.1.9.4 Mesonephros
4.1.10 Thymus
4.1.10.1 Overview
4.1.10.2 Early development of the thymus
4.1.10.3 Cortex and medulla
4.1.10.4 Involution of the thymus
4.1.11 Gut and liver
4.1.12 Bones
4.1.12.1 Vertebral column
4.1.13 Fins
4.1.13.1 Overview
4.1.13.2 Fin anatomy
4.1.13.3 Embryonic fin development (from fertilization to stage 39 [hatching stage])
4.1.13.4 Fin development after hatching (after stage 39)
4.1.13.5 Gene expression during fin development
4.1.14 Gonads
4.1.14.1 Overview
4.1.14.2 PGC specification
4.1.14.3 Formation of gonadal primordium (Figure 4-60b)
4.1.14.4 Sexual dimorphism in germ cell proliferation (Figure 4-61)
4.1.14.5 Posthatching period in XX gonads
4.1.14.6 Posthatching period in XY gonads
4.2 Medaka EGG Envelope and Hatching Enzyme
4.2.1 Overview
4.2.2 Preparation of a hatching enzyme solution from hatching liquid
4.2.2.1 Procedure
4.2.3 Simple method for preparing hatching enzyme solution
4.2.3.1 Procedure
4.2.4 Solubilization of the egg envelope using hatching enzyme
4.3 Observation of Embryos (Embedding Embryos)
4.3.1 Anesthesia of embryos using MS-222
4.3.1.1 Equipment and reagents
4.3.2 Observation of embryos (mounting)
4.3.2.1 Living embryos
4.3.2.2 Processed embryos
4.4 Whole-Mount In Situ Hybridization (see section 4.1.8.4 for a similar protocol)
4.4.1 Fixation and storage
4.4.1.1 Procedure 1
4.4.2 Rehydration, proteinase K treatment, and postfixation at RT
4.4.2.1 Procedure 2
4.4.3 Hybridization and washing
4.4.3.1 Procedure 3
4.4.4 Immunoreaction and washing antibodies
4.4.4.1 Procedure 4
4.4.5 Staining
4.4.5.1 Procedure 5
4.5 Embedding in a Plastic Resin (Technovit 7100)
4.5.1 Equipment and reagents
4.5.2 Agarose mounting (Figure 4-68)
4.5.2.1 Procedure 1
4.5.3 Dehydration and infiltration (Figure 4-68)
4.5.3.1 Procedure 2
4.5.4 Polymerization (Figure 4-68)
4.5.4.1 Procedure 3
References
Chapter 5 Reproductive Behavior of Wild Japanese Medaka
5.1 Wild Japanese Medaka
5.2 Reproductive Behavior of Wild Medaka
5.2.1 Aggressive behavior
5.2.2 Spawning behavior
5.2.3 Egg deposition behavior
5.2.4 Egg discarding behavior
5.2.5 School and aggregation
5.3 Conclusion
References
Chapter 6 Cryopreservation and Transplantation of Medaka Germ Cells
6.1 Introduction
6.2 Cryopreservation of Medaka Testes
6.2.1 Solutions
6.2.2 Materials
6.2.3 Procedures
6.3 Transplantation of Thawed Testicular Cells into Recipient Larvae
6.3.1 Solutions
6.3.2 Materials
6.3.3 Procedures
References
Chapter 7 Genome Editing
7.1 Introduction
7.2 Outline of Targeted Genome Editing Using Nucleases
7.3 Preparation of CRISPR/Cas9 Genome Editing Tools
7.3.1 Materials
7.3.2 Production of custom-designed sgRNA
7.3.2.1 Preparation of the bsai-digested sgRNA backbone
7.3.2.2 Design and production of customized sgRNA
7.3.3 Production of capped RNA encoding a Cas9 nuclease
7.4 Preparation of Custom-Designed TALENs
7.4.1 Materials
7.4.2 Preparation of the TALEN assembly system
7.4.2.1 Preparation of TAL modules (HD1-6, NG1-6, NI1-6, and NN1-6)
7.4.2.2 Preparation of array backbone plasmids (pFUS vectors)
7.4.2.3 Preparation of last repeat modules (LR-HD, NG, NI, and NN)
7.4.2.4 Preparation of TALEN backbone plasmids (pCS2TAL3DD and RR vectors)
7.4.3 Design and construction of custom-designed TALENs
7.4.3.1 Design of TALEN using TALE-NT
7.4.3.2 First assembly: construction of 6-modules array vectors
7.4.3.3 Second assembly: construction of TALEN expression vectors
7.5 Heteroduplex Mobility Assay - A Simple Method to Detect Targeted Genome Modification
7.5.1 Materials
7.5.2 Procedure
7.5.2.1 Identification of the wild type, heterozygotes, and homozygotes
7.5.2.2 Evaluation of the efficiency of targeted genome modifications
7.6 How to Establish Gene Knock-out Strains
7.6.1 Design and synthesis of genome-editing tools
7.6.2 Evaluation of genome-editing activity with fertilized medaka eggs
7.6.3 Microinjection of the selected genome-editing tool(s)
7.6.4 Selection of founder fish by genotyping F1 embryos
7.6.5 Selection of F1 fish carrying the same mutation and the establishment of mutant strain
7.6.6 Selection of homozygous mutant fish in the F2 family
7.7 How to Establish Gene Knock-in Strains
7.7.1 Design and synthesis of CRISPR/Cas9 components
7.7.2 Evaluation of genome-editing activity with fertilized medaka eggs
7.7.3 Construction of donor plasmid with homology arms (Ca. 0.5 kbp) and bait sequences
7.7.4 Microinjection for establishing knock-in strains
7.7.5 Selecting G0 founders harboring the insert gene in the genomic target site
7.A Simple Genomic DNA Preparation by an Alkaline Lysis Method
7.A.1 Materials
7.A.2 Procedure
References
Chapter 8 Photo-Inducible Gene Expression in Medaka
8.1 Outline of IR-LEGO
8.2 Practical Strategies of IR-LEGO in Medaka Study
8.2.1 Selection of heat shock promoters and application studies
8.3 Laser Irradiation Conditions and Sample Preparation
8.4 Caution in Maintaining Strains
8.5 Other Uses of IR-LEGO
8.6 Summary and Future Prospects
References
Chapter 9 Screening and Testing Methods of Endocrine-Disrupting Chemicals Using Medaka
9.1 Applied Toxicity Tests for Endocrine Disruptors
9.2 Detection of Androgenic and Antiandrogenic Chemicals Using Medaka
9.2.1 The formation of papillary processes on anal fin rays as an indicative phenotype for exposure of androgenic and/or antiandrogenic chemicals
9.2.2 Candidate biomarkers for assessing the action of androgenic and antiandrogenic chemicals
9.2.3 Visualization of androgenic and antiandrogenic activity as green fluorescence with spiggin-GFP medaka
References
Chapter 10 Application of the Seawater Medaka Oryzias melastigma (McClelland) for Marine Ecotoxicology
10.1 Background and Development of Oryzias melastigma for Marine Ecotoxicology
10.2 Marine Medaka Developmental Staging
10.3 Standard Breeding and Rearing Conditions
10.3.1 Seawater
10.3.2 Temperature
10.3.3 Photoperiod
10.3.4 Feeding
10.3.5 Embryo collection and rearing
10.3.6 Hatching and larvae collection
10.3.7 Larvae rearing
10.3.8 Larvae feeding
10.4 Raising Marine Medaka for Experimental Use
10.4.1 Experiments using adult fish
10.4.2 Experiments using larvae
10.5 Troubleshooting
10.5.1 Mass mortality
10.5.2 Low egg production
10.5.3 Extensive algal growth
10.6 How to Obtain Marine Medaka O. melastigma
10.7 Experimental Protocols Using Marine Medaka
10.8 Immunotoxicity Assessment: Bacteria Challenge Assays
10.8.1 SOP for adult bacterial challenge assay
10.8.2 SOP for larval bacterial challenge assay
10.8.3 Age selection for larval bacterial challenge
10.9 Fish Dissection and the Whole Adult Histoarray
10.9.1 SOP for fish dissection
10.9.2 SOP for adult medaka histoarray
10.10 Embryo Chip
10.10.1 SOP for embryo and larvae histoarray
Appendix
10.A Materials for SOP for Adult Medaka Histoarray (see section 10.9.2)
References
Chapter 11 Telomerase and Telomere Biology in Medaka
11.1 Introduction
11.2 SOP for Quantification of Telomerase Activity Using the Real-Time Quantitative Telomeric Repeat Amplification Protocol (RTQ-TRAP)
11.2.1 Procedures for sample extraction
11.2.2 Procedures for determination of protein concentration
11.2.3 Procedures for RTQ-TRAP linearity test
11.2.4 Calculation of telomerase activity
11.3 SOP for Quantification of Telomere Length Using Southern Blotting Analysis
11.3.1 Procedures for genomic DNA extraction and digestion with restriction enzymes
11.3.2 Procedures for probe preparation
11.3.3 Procedures for electrophoresis and southern blotting
11.3.4 Procedures for hybridization and detection
11.3.5 Procedures for computerized telomere analysis
11.4 SOP for Quantification of Telomere Length Using Fluorescence In Situ Hybridization
11.4.1 Procedures for fluorescence in situ hybridization
11.4.2 Procedures for confocal microscopy detection
11.4.3 Procedures for ImageJ analysis
References
Chapter 12 Assessments of Medaka Skeletal Toxicity
12.1 Introduction
12.2 Methods
12.2.1 Embryonic exposures: dioxin
12.2.2 Embryonic exposure: dithiocarbamates
12.2.3 Whole-mount Alcian blue staining of hatchlings/larvaea
12.2.4 Whole-mount Alizarin red S staining of hatchlings/larvaec
12.2.5 In vivo Alizarin complexone fluorescent staining for mineralized bone matrix
12.2.6 In vivo calcein fluorescent staining for mineralized bone matrix
12.2.7 Confocal imaging of embryo/hatchling medaka
12.2.8 Morphological assessments
12.3 Results and Discussion
12.3.1 Dithiocarbamates
12.3.2 Dioxin
References
Appendix A Solutions
References
Attributions
Index
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