Over the last two decades, zebrafish have been established as a genetically versatile model system for investigating many different aspects of vertebrate developmental biology. This book is intended for students and everyone who is involved in aquaculture and fisheries. Making gynogenetic diploid zebrafish by early pressure charline walker1, greg s. Methods and protocols, expert researchers provide a collection of experimental techniques for the genetic modification, manipulation, and in vivo live imaging of zebrafish. Zebrafish undergo sexual reproduction, so breeding requires both male and female fish. Research with this model has also expanded into other. The book is extensively illustrated and contains more than 80 figures and 16 tables.
The study of zebrafish as a leading model organism for developmental biology is rapidly expanding. Of these, ssr 12 exhibited no recombinant tetratypes among 175 halftetrad embryos, placing this marker within 1 cm of the centromere of linkage group xvii. Numerous and frequentlyupdated resource results are available from this search. Ten ca n microsatellite simple sequence repeat ssr markers, 1, 2, 12, 14, 16, 18, 20, 22, 26, and 29, were used to show high chiasma interference and to determine centromeremarker map distances in the zebrafish danio rerio. As the disease progresses, fish become lethargic, the fins particularly the dorsal fin are held close to the body, and the fish stop producing eggs. Zebrafish genome yields significant similarity to human genome. Transparent adult zebra fish will make human biology even clearer date. A combination of small size, rapid embryonic development, fecundity, external fertilization, and optical transparency permit the conduct of largescale, forwardgenetic screens. Zhiyuan gong and his colleagues at the national university of singapore were working with a gene that encodes the green fluorescent protein gfp, originally extracted from a jellyfish, that naturally produced.
This volume collects sophisticated and practical techniques, including several not yet widely known, from laboratories around the world. It involves carrying a baby inside you for nine months, caring for a child for a number of years, and many more. Feb, 2020 f1 fish are interbred and the f2 progeny, referred to as an f2 family, are raised to adulthood. The zebrafish genome has been fully sequenced, which is a musthave for model organisms nowadays. Method for producing gynogenetic fishes with diagram. Zebrafish are amenable to such studies because the generation time of zebrafish is short, adults are small and easy to maintain, and females typically lay up to several hundred eggs per mating.
According to lestage 1933, the weak electrical currents if. Although adult fish are not suitable for highthroughput in vivo drug screening, zebrafish embryos are uniquely suited for this purpose kaufman et al. Alternately, fish can be obtained from carolina biologicals 8003345551. Generation of clonal zebrafish line by androgenesis without.
New conditions for generation of gynogenetic halftetrad. Parthenogenesis in sharks has been confirmed in at least three species, the bonnethead, the blacktip shark, and the zebra shark, and reported in others. Production of clones of homozygous diploid zebra fish brachydanio rerio. Students demonstrate an understanding of how living systems function and how they interact with the physical environment. Embryo viability was significantly reduced in inbred exposed fish and there was a tendency for inbred males to sire fewer offspring.
Gynogenesis is production of all female offspring by inactivating sperm. Prior to planning pregnancy, you should learn more about the things involved in getting pregnant. Liang,1 and jacqueline morris department of biology, queens college, and the graduate center, city university of new york, new york 10016, department. A small, plastic plant is included and provides good spawning substrate. Diseases caused by protozoans are common problems in aquarium fishes, and several have been observed in zebrafish. Zebra fish, any member of either of two unrelated groups of fishes, the freshwater species in the genus brachydanio family cyprinidae. Incorporation of a subacute test with zebra fish into a hierarchical system for evaluating the effect of toxicants in the aquatic environment.
Ongoing genetic screens have identified over 7,000 mutations in zebrafish affecting genes that regulate developmental patterning, organogenesis, physiology and behavior. Most gynogenetic species fall under the taxonomic groups of fish and amphibian amazon mollies, aka poecilia formosa require the sperm of closely related male poecilia latipinna to engage in gynogenesis. One of the practical goals of induced gynogenesis is the production of genetically identical population, i. Breeding plans, mass seed production and aquaculture techniques. Overview zebrafish danio rerio has emerged recently as a premiere organism to study vertebrate development and genetics. Discussion it is important to make sure that embryos produced by this method are true gynogenetic diploids. Synaptonemal complex karyotype of zebrafish heredity. Halftetrad gynogenesis can be accomplished using hydraulic pressure to disrupt microtubulemediated segregation during meiosis ii, which normally occurs after fertilization. A highquality sequence assembly of the zebrafish genome reveals the largest gene set of any vertebrate and provides information on key genomic features, and comparison to. Ichthyophthirius multifiliis the cause of ich or white spot disease and piscinoodinium pillulare the cause of freshwater velvet disease are very common in warm water captive fishes. Production of clones of homozygous diploid zebra fish brachydanio rerio nature. Zebra fish make a perfect candidate for embryology demonstrations or labs in the classroom setting.
The zebrafish danio rerio has been extensively used in biomedical research as a model to study vertebrate development and hematopoiesis and recently, it has been adopted into varied fields including immunology. In most species, gonads are paired organs of similar size, which can be partially or totally fused. This effort showed that their genome is remarkably similar to the human one, with at least 70% of human genes having a zebrafish equivalent a figure that is even higher when only diseasecausing genes are considered. However, conroy and conroy 1999 reported that kanamycin at 50 ppm with 4 doses 2 days apart apparently controlled the disease in guppies. Brachydanio rerio definition of brachydanio rerio by. Induced development of triploids and diploid gynogenetic and androgenetic fish includes 1 to 10 min exposure of activated eggs to hhp of 48. Jul 01, 2016 in the last 30 years, the zebrafish has become a widely used model organism for research on vertebrate development and disease.
A primer on the vertebrate model danio rerio nathalia g. The oldest domesticated fishes, and the consequences of an. All mature homozygous gynogenetic zebra fish obtained n9 were males. In this article we will discuss about methods for producing gynogenetic fishes.
There are large numbers of ways available for the initiation of gynogenesis in the development of embryo. We have examined the effects of tribulus terrestris tt, a nontoxic herb, on sex reversal in the convict cichlid cichlasoma nigrofasciatum with the aim of introducing a new environmentally friendly method for masculinization in c. Sequencing of the entire genetic makeup of the zebrafish has revealed that 70 per cent of proteincoding human genes are related to genes found in the zebrafish and that 84 per cent of genes known to be associated with human disease have a zebrafish counterpart. Pdf generation of clonal zebrafish line by androgenesis. This includes an understanding of the cycling of matter and flow of energy in living systems. Some important stages are in gynogenetic production which is uv irradiation, shortterm storage of sperm, heat shock time and duration, karyotype analysis and morphological screening were applied. Environmental epigenetics in zebrafish epigenetics. Zebrafish represent a vertebrate model organism that has been widely, and increasingly, employed over the last decade in the study of developmental processes, wound healing, microbehost interactions, and drug screening. Zebrafish genetics biology bibliographies cite this for me. A guide for the laboratory use of zebrafish danio rerio 4th ed. An understanding of the characteristics, structure and function of cells, organisms and living systems will be developed. Making gynogenetic diploid zebrafish by early pressure core. Periodic mating keeps eggs healthy, but making all those eggs takes a lot of work, so individual females should only be mated once per week. Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome.
Of the currently available zebrafish tumor models, only two manifest in the embryonic to larval stage so that highthroughput screening in 96well plates is feasible. In the last 20 years, research activity using the zebrafish danio rerio has increased dramatically, due in part to the ease of breeding and raising them, their genetic tractability, embryonic. Gynogenetic definition of gynogenetic by the free dictionary. Zebrafish genetics biology bibliographies in harvard style. Pdf making gynogenetic diploid zebrafish by early pressure. Microsatellitecentromere mapping in the zebrafish danio. Zebrafish genome yields significant similarity to human.
The classical method is to prick each ovum with a needle dipped in serum of blood. Acknowledgments methods for in vitro fertilization and gynogenetic diploids were developed for zebrafish by charline walker in george streisingers lab at the university of oregon in. Method for creating a uniform vascular wound in zebrafish or. With the credentials of zebrafish as a developmental model now well recognized, the emerging new opportunity is the wider application of zebrafish biology to aspects of human disease modelling. Fish with velvet disease have the characteristic behavior of rubbing their sides and flipping around in the corners of the tank. Published by habiba akter hussain 55 kristawpur, mymensingh 2200 bangladesh isbn 9843218396 cover and other photos by m. We take advantage of zebrafish as a model system for development and genetics, of stickleback as a model for evolutionary biology and evo devo, and of teleosts more generally, the most species rich of all vertebrate groups, for their outstanding phenotypic variation. The original zebrafish or zebra danio, danio rerio is a native of rivers in india and bangladesh.
Generation of clonal zebrafish line by androgenesis without egg irradiation. After fertilization, larvae survive with only the innate immune responses because adaptive immune system is morphologically and functionally mature only after 46 weeks. Fish reproductive organs include testes and ovaries. A few recent studies have suggested that pollutant exposure could induce alteration in dna methylation patterns by disturbing dnmt gene expression during zebrafish embryogenesis. Making gynogenetic diploid zebrafish by early pressure. The zebrafish model system has many attributes that contribute to its utility in identifying genes required for hematopoiesis. The oldest domesticated fishes, and the consequences of an epigenetic dichotomy in fish culture. Each f2 family contains two mutagenized genomes, one from its mother and one from its father. The generation of gynogenetic halftetrads is an important tool for genetic mapping and mutant screens in zebrafish danio rerio. Once anesthetized, gently blot fish dry on paper towel, paying special attention to dry ventral side.
Excess water will swell the eggs and prevent fertilization. Forward screens of ep diploid embryos are an efficient way to identify genes involved in both early and later developmental processes and are being used today 5,7. It measures three centimeters long and has gold and dark blue stripes. Central divider allows for separation of sexes until breeding is desired. Transparent adult zebra fish will make human biology even. Tt is a natural plant product that elevates the testosterone levels in humans and animals. Position fish on sponge holder, ventral side up fig. The zebrafish book a guide for the laboratory use of zebrafish danio brachydanio rerio by monte westerfield, institute of neuroscience, university of oregon older literature referred to zebrafish as brachydanio rerio. Different heat shock application effect on gynogenetic. Zebrafish are common aquarium fish and are relatively easy to maintain. Observations of fish eating and maintenance behavior in two species of basiliscus pp.
Zoology and wildlife conservation biological sciences aquaculture research fishculture goldfish physiological aspects. The latter is recognized as a cause of disease in zebrafish held in research facilities. Among these, a couple of reports described the impact of 2,3,7,8tetrachlorodibenzopdioxin tcdd on dna methylation of embryos, larvae, and adult zebrafish 92, 93. Katsutoshi arai, in reproductive biotechnology in finfish aquaculture, 2001. Completely paired scs from both spermatocytes and oocytes were measured to.
The growing interest in zebrafish research was paralleled by an increase in tools and methods. One of the most significant features of the zebrafish system is the ability to conduct largescale forward genetic screens in an organism with high genetic conservation to the human genome. The zebrafish as a model for cancer pubmed central pmc. However, the equipment used in the original studies is no longer available, and the. In the last 20 years, research activity using the zebrafish danio rerio has increased dramatically, due in part to the ease of breeding and raising them, their genetic tractability, embryonic accessibility, and their imaging potential. Reliable information about the coronavirus covid19 is available from the world health organization current situation, international travel. Production of cloned fish by chromosome manipulation and. In a zygotic screen, the fish of a given f2 family are intercrossed, and the f3embryos are screened for recessive mutant defects. Oclcs webjunction has pulled together information and resources to assist library staff as they consider how to handle coronavirus. Danio rerio is a tropical fish native to india, bangladesh, nepal, and myanmar.
The gynogenetic fish producing method and the different heatshock applications mentioned in this study enhanced the survival rate of gynogenetic fish. Zebrafish are remarkably biologically similar to people and share the majority of the same genes as humans, making them an important model for understanding how genes work in. Meiotic cells of zebrafish have been prepared to show synaptonemal complexes scs by light and electron microscopy. Zebrafish is an important model organism in areas of embryogenesis, genetics and regeneration. The infection is usually very difficult to eradicate in fish culture systems with antibiotics. There may also be a range of secondary organs that increase reproductive fitness. A bonnethead, a type of small hammerhead shark, was found to have produced a pup, born live on 14 december 2001 at henry doorly zoo in nebraska, in a tank containing three female hammerheads.
Through a powerful combination of genetics and experimental embryology, significant inroads have been made into the regulation of embryonic axis formation, organogenesis, and the development of neural networks. The genital papilla is a small, fleshy tube behind the anus in some fishes, from which the sperm or eggs are released. Students will also develop a deeper understanding of the. A fresh look at zebrafish from the perspective of cancer. A guide for the laboratory use of zebrafish brachydanio rerio.
Exposure of zebra fish to a chemical environmental agent, analogous to that caused by anthropogenic pollution, amplified the effects of inbreeding on key reproductive traits. Streisinger g, walker c, dower n, knauber d, singer f 1981 production of clones of homozygous diploid zebra fish brachydanio rerio. Mesabtricaine diluted in fish water recipe in the zebrafish book 3. Introduction gynogenesis is a special form of reproduction that has been known to occur in nature. Gynogenesis presented by shrish chandra yadav fgb0517 college of fisheries, cau lembucherra, agartala 2. The use again of streisingers zebrafish cloning methods to produce fully homozygous gynogenetic doubledhaploid zebrafish from the ab and the tubingen tu strains, followed by the production and sequencing of 459 of their f2 progeny for 201,917 snp loci produced the satmap, with the greatest resolution of any animal genetic mapping panel. Males have slimmer bodies with a slightly red hue, while females have bigger, silver bellies full of eggs. Making gynogenetic diploid zebrafish by early pressure protocol. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow f1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. It is a popular aquarium fish which is frequently sold under the trade name zebra danio. To establish clonal lines in fish, the first step is to obtain the first generation of doubled haploids dhs, which are completely homozygous gynogenetic or androgenetic diploids generated by inhibition of the first cleavage event to produce a double chromosome set after parthenogenetic development. A highthroughput sperm freezing protocol for zebrafish. Using genalex, all 30 fish were correctly assigned to their line except for three. These fish showed characteristics as described by wixon 2000.
Powerful techniques allow efficient generation and recovery of zebrafish mutations. The zebrafish has become a popular model system for genetic studies of vertebrate development. Water activates sperm so it is important to thoroughly dry around the cloacae. Methods for in vitro fertilization and gynogenetic diploids were developed for zebrafish by charline walker in george streisingers lab at the. A plex of tetra and pentastrs to identify zebrafish. Zebrafish genome found strikingly similar to humans. The optical transparency of zebra fish embryos is especially suited for investigations into hematopoiesis and stem cell biology. Research has shown that the males prefer to mate with other female poecilia latipinna, due to the previously discussed disadvantage for males in mating with gynogens. A guide for the laboratory use of zebrafish danio rerio.