Medaka: Biology, Management, and Experimental Protocols

Contents......Page 7
Contributors......Page 19
Preface......Page 23
1.1 History......Page 27
1.2.1 Phylogeny and distribution of medaka and relatives......Page 29
1.2.2 Genetic diversity of medaka......Page 33
1.3.1 Advantageous features in general......Page 35
1.3.2.1 Introduction and history......Page 37
1.3.2.2 Body color and chromatophores......Page 38
1.3.2.3 Genes mutated in body-color mutants......Page 40
1.3.3 Wild strains......Page 42
1.3.4.1 History for establishing inbred lines......Page 44
1.3.4.2 Characteristics of medaka inbred strains......Page 45
Column 1.1 For those who cannot decide which medaka to use......Page 46
1.3.4.4 To generate and maintain medaka inbred strains......Page 47
Column 1.2 Variation among strains......Page 48
1.3.5 Differences from zebrafish......Page 49
2.1.2 Contact the National Bio-Resource Project (medaka) in Japan......Page 57
2.1.4 Catching medaka from the wild......Page 58
2.2.1 Breeding program......Page 59
2.2.2.1 Aquarium system......Page 61
2.2.2.2 Maintenance of recirculating system......Page 66
Column 2.1 Soft water is suitable for medaka breeding......Page 67
2.2.3.2 Water system at JST Kyoto facility......Page 73
2.2.3.3 Water condition......Page 74
2.2.4.3 Daily care......Page 75
2.2.5 Outdoor breeding......Page 76
2.3.2 Feeding schedule......Page 80
2.3.3.1 Brine shrimp (Artemia)......Page 81
2.3.3.2 Dry feed......Page 85
2.4.1 Tail rot disease......Page 86
2.4.2 Matsukasa disease......Page 87
2.4.3 Trichodina......Page 88
2.4.5 White spot disease......Page 89
2.4.7 Gyrodactylus......Page 90
2.5 Goods for Medaka......Page 91
3.1.1 Sex determination in medaka......Page 93
3.1.2 Sex determination in the genus Oryzias......Page 94
3.2.1 Hypothalamic–pituitary–gonadal axis......Page 95
3.2.2 Oocyte growth and maturation......Page 96
3.2.3 Spermatocyte growth and maturation......Page 97
3.3.1 Oogenesis......Page 98
3.3.2 Spermatogenesis......Page 100
3.4.1 Morphology and biochemical characters of the medaka egg envelope......Page 101
3.4.2 Origin of the egg envelope in medaka fish......Page 102
3.4.3 Gene structure of egg envelope glycoproteins in medaka......Page 106
3.4.4 Molecular mechanisms of liver-specific expression of Choriogenins’ Genes......Page 107
3.4.5 Assembly of the Choriogenins into the egg envelope in the ovary......Page 110
3.4.7 Conclusion......Page 112
3.5 Necessary Conditions for Spawning......Page 113
3.6 Reproductive Behavior......Page 114
3.7 Mating......Page 115
3.9.1 Cleaning......Page 116
3.11 Generation of Sex-Reversed Medaka......Page 117
3.11.3 High-temperature treatment to generate XX males......Page 118
Column 3.1 Interstrain Variation in Reproductive Performance......Page 119
4.1.2 Sorting of eggs or fish......Page 127
4.1.5 MTA (Material Transfer Agreement)......Page 128
4.2.2 Procedure (Movie M4-1)......Page 130
4.3.2 The procedure for cryopreservation (Figure 4-2 and Movies M4-2)......Page 131
4.3.3 Materials......Page 132
4.3.5 Procedures......Page 133
4.4.1 Overview......Page 136
4.4.2 Solutions......Page 137
4.4.3 Materials......Page 138
4.4.4 Procedures (Movies M4-3)......Page 139
Column 4.1 Infertile mating method for collecting unfertilized eggs......Page 141
5.1.1 Secondary sexual characters......Page 143
5.1.2.1 Pigment cells (chromatophores)......Page 144
5.1.2.3 Chromatophores in medaka......Page 145
5.1.2.4 Chromatophore distribution in medaka......Page 149
5.2.1.1 Observations of internal organs in the live see-through medaka......Page 150
5.2.1.2 Dissection of adult medaka......Page 152
5.2.3.1 Adult central nervous system......Page 153
5.2.3.2 Adult peripheral nervous system......Page 165
5.2.4.1 Hypothalamo–pituitary system......Page 173
5.2.4.2 Pineal organ (epiphysis)......Page 176
5.2.4.3 Thyroid gland......Page 178
5.2.4.6 Gonads......Page 179
5.2.4.10 Corpuscle of Stannius......Page 180
5.2.5 Gonads......Page 181
5.2.5.1 Ovary......Page 182
5.2.6 Kidney......Page 183
5.2.6.3 Histology of the kidney......Page 184
Column 5.1 How to make sections of a meture ovary for histological analysis......Page 186
6.1.1.0 Stage 0; Unfertilized Egg – Figure 6-1......Page 191
6.1.1.3 Stage 3; two-cell stage (1 hour 5 minutes) – Figure 6-1......Page 193
6.1.1.10 Stage 10; early blastula stage (6 hours 30 minutes) – Figure 6-2......Page 194
6.1.1.16 Stage 16; late gastrula stage (21 hours) – Figure 6-3......Page 196
6.1.1.17 Stage 17; early neurula stage (1 day 1 hour) – Figure 6-3......Page 197
6.1.1.23 Stage 23; 12-somite stage (1 day 17 hours) – Figure 6-4......Page 198
6.1.1.24 Stage 24; 16-somite stage (1 day 20 hours) – Figure 6-5......Page 200
6.1.1.29 Stage 29; 34-somite stage (3 days 2 hours) – Figure 6-5......Page 201
6.1.1.33 Stage 33; stage at which notochord vacuolization is completed (4 days 10 hours) – Figure 6-6......Page 202
6.1.1.39 Stage 39; hatching stage (9 days) – Figure 6-7......Page 204
6.1.1.40 Stage 40; first larval stage – Figure 6-8......Page 205
6.1.2 Brain......Page 206
6.1.2.2 Neurula step (stages 17–18)......Page 208
6.1.2.4 Neural tube step (stages 23–27)......Page 210
6.1.2.5 Late embryonic brain step (stages 28–34)......Page 211
6.1.3 Hatching gland......Page 213
6.1.3.1 Origin of fish hatching gland cells......Page 214
6.1.3.2 Secretion of hatching enzymes from hatching gland cells......Page 216
6.1.4.2 Eye field determination and establishment of retinal identity......Page 217
6.1.4.3 Splitting of the retinal anlage into two retinal primordia......Page 219
6.1.4.5 Morphogenesis II: formation of the optic cup......Page 220
6.1.4.8 Retinotectal projection......Page 222
6.1.5.1 Skeletal development......Page 224
6.1.5.2 Muscle development......Page 225
6.1.6.1 Vascular anatomy of the developing medaka......Page 227
6.1.6.2 Origin of the medaka endothelial lineage......Page 236
6.1.6.3 Abbreviations......Page 237
6.1.7.2 Observation of Embryonic and Adult Blood Cells......Page 238
6.1.8.1 Overview......Page 240
6.1.8.2 Heart architecture......Page 241
6.1.8.3 Heart morphogenesis......Page 242
6.1.8.4 Observation of the developing heart......Page 251
6.1.9.2 Nephrogenesis......Page 253
6.1.10.2 Early development of the thymus......Page 255
6.1.11 Gut and liver......Page 257
6.1.12.1 Vertebral column......Page 260
Column 6.1 Key words in bone formation......Page 268
6.1.13.2 Fin anatomy......Page 269
6.1.13.4 Fin development after hatching (after stage 39)......Page 270
6.1.13.5 Gene expression during fin development......Page 272
6.1.14.2 PGC specification......Page 273
6.1.14.4 Sexual dimorphism in germ cell proliferation (Figure 6-61)......Page 275
6.1.14.6 Post-hatching period in XY gonads......Page 277
6.2.1 Overview......Page 278
6.2.2 Preparation of a hatching enzyme solution from hatching liquid......Page 280
6.2.4 Solubilization of the egg envelope using hatching enzyme......Page 281
6.3.1 Anesthesia of Embryos using MS-222......Page 282
6.3.2.1 Living embryos......Page 283
6.3.2.2 Processed Embryos......Page 286
6.4.1 Fixation and storage......Page 287
6.4.2 Rehydration, proteinase K Treatment and post-fixation at RT......Page 288
6.4.4 Immunoreaction and washing antibodies......Page 289
6.5.1 Agarose mounting (Figure 6-68)......Page 290
6.5.3 Polymerization (Figure 6-68)......Page 291
Column 6.3 Pigment cells (Figure 6-69)......Page 292
Column 6.4 Kupffer’s vesicle......Page 293
7.1 Microinjection Technique for Medaka Eggs......Page 303
7.1.1.1 Egg holder......Page 304
7.1.1.2 Glass needles for microinjection......Page 306
7.1.1.3 Injector and manipulator with needle holder......Page 307
7.1.1.4 Microscope and light......Page 308
7.1.2 Microinjection procedure......Page 309
Column 7.1 Microinjection into nuclei......Page 311
7.2.1 DNA microinjection for transgenesis and transient expression......Page 313
7.2.2 DNA construction for transgenesis......Page 315
7.3 RNA Microinjection......Page 317
7.4 Gene Knockdown Technology......Page 318
7.4.2 gripNAs......Page 319
8.1 Status of Medaka in Toxicology......Page 323
8.2.1 Preparation and acclimation of fish......Page 324
8.2.2 How to expose to chemicals......Page 327
8.3.2 Acute Toxicity Test (OECD TG203)......Page 329
8.3.3 Early-life stage toxicity test (OECD TG210)......Page 330
8.4 Applied Toxicity Tests for Endocrine Disrupters......Page 331
8.4.2 Fish full lifecycle testing (FFLC) using medaka......Page 332
8.4.3 Sensitive period to estrogen substances in early life stages......Page 333
8.5.2 Vitellogenin measurement......Page 336
8.5.3 Summary and comments......Page 339
8.6 New Techniques and Other Studies......Page 340
Column 8.1 Application of medaka and olyzias Sp. in seawater. Can medaka survive in seawater?......Page 341
9.1 Medaka Genome Project and Genome Sequence Database......Page 345
9.1.1 Genome database......Page 346
9.1.2 Polymorphism between the Southern and Northern Japanese populations......Page 349
9.2.1 EST database......Page 350
9.2.2 Developmental Expression database......Page 352
9.3.1 Mapping mutants using SSLP and RFLP markers......Page 353
9.3.1.1 Creating mapping panel......Page 356
9.3.1.2 Identification of the linkage group linked to a mutation using bulk segregation analysis with M markers.......Page 357
9.3.1.3 Low-resolution mapping......Page 362
9.3.1.4 Intermediate-Resolution Mapping......Page 364
9.3.1.6 In silico chromosome walking......Page 365
Column 9.2 Construction of fosmid library......Page 369
10.1 Cell Culture from Medaka Embryo......Page 371
10.1.1 Flow chart of primary cell culture from medaka embryo......Page 372
10.1.3 Protocol......Page 373
10.1.4 Notes......Page 374
10.2 In Vitro Spermatogenesis from Primary Spermatocytes......Page 376
10.2.2 Required equipment and materials for primary culture of primary spermatocytes......Page 377
10.2.3 Protocol......Page 378
10.3 Single Cell Labeling......Page 379
10.3.1 Flow chart of single cell labeling......Page 380
10.3.3 Protocol of single cell labeling......Page 381
10.4 Imaging of Living Embryos......Page 383
10.4.2 Fluorescent labeling......Page 385
10.4.5 Data analysis......Page 386
10.4.7 Conclusion......Page 387
10.5.1 Cell transplantation in embryo. Figure 10-11 shows the procedure of cell transplantation in embryo briefly......Page 388
10.5.2 Scale......Page 390
10.6 Nuclear Transplantation......Page 393
10.6.1 Equipment and materials......Page 394
10.6.3 Perspectives......Page 395
10.7.2 Mutagens that have been used for medaka......Page 396
10.7.3 Mutagenesis screen using ENU......Page 397
10.8 Tilling (Gene Knockout)......Page 399
10.8.1 Outline of the TILLING method......Page 401
10.8.3 About SNPs......Page 406
10.8.4 How to obtain a Medaka TILLING library......Page 407
10.9 Cell Trace Experiment with a Caged Fluorescent Dye During Medaka Gastrulation......Page 408
10.9.2 Equipment and materials......Page 410
10.9.3 Protocol (Figure 10-2)......Page 411
10.9.4 Notes......Page 412
Appendix 1 Guidelines on Using Medaka in Experiments......Page 415
Appendix 2 Internet Websites Related to Medaka Research......Page 417
Appendix 3 Solutions......Page 423
Appendix 4 Inbred Strains, Closed Colonies, and Mutant Strains......Page 425
Appendix 5 Index of Abbreviation......Page 429
Attributions......Page 433
Index......Page 437