FrontPage

Medaka Book†

Preface†

Contents†

1. Keeping medaka
   • 1.1 tanks
     • 1.1.1 tanks used in Nagoya Univ.
     • 1.1.2 cleaning tanks
   • 1.2 water
     • 1.2.1 water in Nagoya
     • 1.2.3 checking water quality
   • 1.3 food
     • 1.3.1 artificial food
   • 1.4 disease
   • 1.5 room conditions---temperature, illumination, etc.
2. Breeding medaka
   • 2.1 embryo collection
     • 2.1.1 simple method for obtaining next generation
     • 2.1.2 artificial insemination
     • 2.1.3 sterilization of embryos by bleaching Under construction
   • 2.2 embryo culture
   • 2.3 preparation of food for medaka
     • 2.3.1 culture of paramecia
     • 2.3.2 culture of brine shrimp
3. Maintaining medaka stocks
   • 3.1 inbred lines
   • 3.2 closed colonies
   • 3.3 freezing medaka sperm
     • 3.3.1 Cryo-preservation of Medaka sperm
     • 3.3.2 Cryopreservation of Medaka Spermatozoa (trehalose buffer)
     • 3.3.3 Artificial insemination using frozen sperm
4. Looking inside of medaka
   • 4.1 anatomy
   • 4.2 sectioning
     • 4.2.1 Frozen sectioning
   • 4.3 3D imaging
   • 4.4 Skeleton
     • 4.4.1 Staining of cartilage and bone in whole embryos, larvae and adults by Alcian blue and Alizarin red
     • 4.4.2 Calcein and Alizarin complexone staining
     • 4.4.3 von Kossa staining of sections
     • 4.4.4 TRAP staining of sections
   • 4.5 Blood vessel
     • 4.5.1 Alkaline phosphatase staining
     • 4.5.2 Confocal microangiography
     • 4.5.3 Dye injection
5. Culturing cells
   • 5.1 cell culture
     • 5.1.1 primary cell culture from the caudal fin
   • 5.2 chromosomal analysis
     • 5.2.1 chromosomal analysis of primary cultured cells from the caudal fin
   • 5.3 FISH
6. Looking at gene expression in medaka
   • 6.1 preparation of hatching enzyme
     • 6.1.1 preparation of hatching enzyme (Nagoya)
   • 6.2 in situ hybridization
     • 6.2.1 Whole-mount in situ hybridization (Inohaya version)
     • 6.2.2 In situ hybridization in fins
   • 6.3 two-color in situ hybridization
   • 6.4 immunohistochemistry
     • 6.4.1 Antibody staining in fins
7. Generating transgenic medaka
   • 7.1 construct of transgene
   • 7.2 microinjection
   • 7.3 isolation of a transgenic line
8. Screening a new mutant---Mutagenesis
   • 8.1 ENU mutagenesis
   • 8.2 specific locus test
   • 8.3 screening
   • 8.4 complementary test
9. Manipulating gene expression in medaka
   • 9.1 ectopic expression of genes by DNA or RNA injection
     • 9.1.1 DNA construct
     • 9.1.2 preparation of synthetic RNA
   • 9.2 knock-down of genes by antisense morpholino oligonucleotide injection
   • 9.3 transplantation
     • 9.3.1 Labeling of embryo and transplantation of the embryonic shield
10. Mapping and cloning
    • 10.1 mapping a gene
      • 10.1.1 preparation of mapping panel
      • 10.1.2 detection of polymorphism
    • 10.2 determination of linkage group---M-marker analysis
      • 10.2.1 genomic DNA extraction
      • 10.2.2 PCR amplification of M-markers from the DNA pools
      • 10.2.3 separation of PCR products and determination of the candidate linkage group of mutation
    • 10.3 choromosome walking
11. Histology
    • 11.1 histochemistry
      • 11.1.1 PAS staining
      • 11.1.2 Acridine orange staining
      • 11.1.3 TUNEL staining
12. Exposing medaka to chemicals
13. Getting tools and facilities
14. How to get information

Editors†

Guide to authors†

Citation†