The zebrafish information network zfin is an online database that collects, curates, and disseminates information about zebrafish biology. Zebrafish get their name because they resemble zebras. The zebrafish danio rerio is a freshwater fish belonging to the minnow family of the order cypriniformes. The zebrafish has an equivalent anatomical site to mammalian blood islands, known as the intermediate cell mass icm, where two stripes of mesodermal cells expressing tal1, lmo2, and gata1a converge to the midline of the zebrafish embryo and are surrounded by endothelial cells that become the cardinal vein aladhami and kunz, 1977, detrich. Zebrafish are able to efficiently regenerate lost cardiac muscle, and is used as a model to understand why natural heart regeneration is blocked in mammals. Quantification of estradiol uptake in zebrafish embryos and. Zebrafish are an effective model of appendage regeneration. Effects of lethal irradiation in zebrafish and rescue by. Firstly, a zebrafish alcoholic liver injury model was established using1. All efforts were made to minimize suffering, using 0.
Includes content submitted by the community and imported from the zebrafish book, 5th edition. Zirc frequently asked questions zebrafish international. Background elucidating the complex cell dynamics divisions, movement, morphological changes, etc. Guidelines for use of zebrafish in the nih intramural research program. Briefly, larvae were anesthetized in 160 mgml tricaine sigmaaldriche3 medium and injected with a total of 1 nl gentamicin 812 ng sigmaaldrich diluted in saline into the common cardinal vein. Make up a 15 mm stock concentration of tricaine ethyl 3aminobenzoate methanesulfonate salt as instructed in the zebrafish book 23. Sigmaaldrich, and microinjected with the labeled tumor cells into the subperidermal space of the yolk sac 8, 18. Oct 27, 2009 taken from the book zebrafish, edited by christiane nussleinvolhard and ralf dahm, page 22. Cyprinidae, which contains over 2,400 species, including the loveable goldfish. A twocolor acidfree cartilage and bone stain for zebrafish larvae. Hardcopies of the 4th edition of the zebrafish book can be obtained for a nominal fee from the zebrafish international resource center, 5274 university of oregon, eugene, or 97403 usa. Development of zebrafish danio rerio as a natural model. Ann arbor, mi usa stock by dissolving 200 mg tricaine pow.
Cellular dissection of zebrafish hematopoiesis sciencedirect. Finquel, ms222, tricaine s is used for anesthesia or euthanasia of fish and other coldblooded aquatic animals. An optimized method for counting dopaminergic neurons in. Zebrafish are native to the southeastern himalayas and thought to originate from the ganges. Adrenocortical carcinoma xenograft in zebrafish embryos as. Zebrafish eggs were collected, sorted to remove dead and unfertilized embryos, and placed in. Zebrafish embryos were kept in a 28 c incubator in e3 media 5 mm nacl, 0.
Ethanol treatment zebrafish eggs were collected, sorted to remove dead and unfertilized embryos, and placed in 24well culture plates at a density of nov, 2008 using standard body length as a measure of growth of fish reared in isolators as described 26, we find that the average body length of gf zebrafish at 14 dpf 4. Zebrafish embryos and larvae were handled in strict accordance with good animal practice as approved by the appropriate committee mdi biological laboratory animal core iacuc number 20. Eyes were dissected from animals younger than a month, and lenses were dissected 9 from animals older than a month. Zebrafish by sharon emerson meet your next favorite book. Immediately after confirming the lack of operculum movement, the fish were positioned dorsal side up on a piece of ice covered by a wiper wypall x60. Ucsd guidelines for the preparation, storage and use of tricaine methanesulfonate. Zebrafish is a cute book with fun illustrations and a great message. O in e3 media for 20 minutes and wash it in e3 media for 5 minutes.
Recently, zebrafish has sparked interest in other branches of the biomedical research. Mar 12, 2020 this is where zebrafish researchers can share experimental protocols and tips with the rest of the research community. The zebrafish is a widely used model organism throughout many aspects of science, the applications of which are growing 6. How best to immobilise zebrafish larvae for imaging. The zebrafish is a tropical fish native to southeast asia. Zebrafish stock photos download 669 royalty free photos. Anesthetize adult zebrafish in tricaine added to the swimming water to immobilize them.
Thus tricaine may inhibit the zebrafish heart rate by blocking sodium ion permeability in the early embryos before machr appearance. When zebrafish caudal fins tail fin are amputated, epithelial cells migrate to repair the injury, creating an apical epidermal cap. Finquel, ms222, tricaines is used for anesthesia or euthanasia of fish and other coldblooded aquatic animals. Klionsky life sciences institute and departments of molecular, cellular and developmental biology. However, the utility of zebrafish for modeling adultonset heart disease has been limited by a lack of robust methods for in vivo evaluation of cardiac function. Scn5ad1275n transgenic zebrafish blue columnslines. Download 669 zebrafish stock photos for free or amazingly low rates.
Zebrafish danio rerio as a vertebrate model host to. Welcome zebrafish in the classroom is designed to be a resource for teachers and students who are using zebrafish in undergraduate courses. An adult zebrafish was anesthetized with tricaine and had a tail clip taken for genomic analysis with a sharp razor blade. It is a blue fluorescent dye that binds nucleic acids both by groove binding.
The zebrafish danio rerio is an increasingly popular model organism in cardiovascular research. Zebrafish heart regeneration occurs by cardiomyocyte. The procedure for removing the chorions can be found in the zebrafish book or at. Adult fish that require euthanization due to infertility, age, or experimental need should be anesthetized with lethal dose of tricaine 200300 mgl in system water or e3 solution containing 5 mm nacl, 0. Standard operating procedures chen laboratory zebrafish facility. The bones of euthanized adult zebrafish were stained using aliziran red sigma and the cartilage with alcian blue sigma 18. Biology, husbandry, diseases, and research applications is a comprehensive work that fulfills a critical need for a thorough compilation of information on this species. Poke the larvae with a capped microloader pipette tip to assess their touch response before proceeding. Detail protocols for fish can be obtained from a number of practical books but they. To satisfy this criterion, we developed a transgenic zebrafish line, called photo, that allows photoconvertible optical tracking of nuclear and membrane. Zebrafish 2535 days old were immunosuppressed with 10. Zebrafish danio rerio zebrafish are small teleost freshwater fish of the carp family, named after the five horizontal blue stripes extending from gills to caudal fin tip. The text provides significant updates for working vivarium professionals maintaining zebrafish colonies, veterinarians responsible for their care and wellbeing, zoologists and ethologists.
Welcome the same characteristics that make zebrafish a powerful system for basic research also make them an excellent tool for teaching. Zebrafish ab strain were raised and maintained under standard laboratory conditions. The employment of the zebrafish model system in the study of kidney diseases like aki has led to important observations and will continue to be a good model to study kidney injury 7,8. Historically, zfin hosted the zirc site and databases. Anesthesia and euthanasia in zebrafish oxford academic journals. Immobilization by submersion in ice water 5 parts ice1 part water, 04. Zebra fish definition is any of various barred fishes. Male adult zebrafish 3months postfertilization were anesthetized with 1. Louis, mo added to the circulating water of the breeding system. Zebrafish are small teleost freshwater fish of the carp family, named after the five horizontal blue stripes extending from gills to caudal fin tip. A guide for the laboratory use of zebrafish danio rerio westerfield, monte on. Eighteen male zebrafish abwt, date of fertilisation.
Reynolds is the first graphic novel from the popular fablevision publisher. Using acridine orange to measure cell death in ethanol. L of 1% evans blue dye into the sinus venosus with a 30gauge needle attached to a 0. Zebrafish, so named due to their stripes, prefer to live in large groups called. Quantification of estradiol uptake in zebrafish embryos. Optical development in the zebrafish eye lens wang. The use of zebrafish danio rerio as a model organism began in the 1960s. Stauffer and colleagues 1988 acclimated blue tilapias. Male adult zebrafish were anesthetized with tricaine 3. This is where zebrafish researchers can share information with the rest of the community.
A guide for the laboratory use of zebrafish danio rerio, eugene, university of oregon press. A transgenic zebrafish model of a human cardiac sodium. A guide for the laboratory use of zebrafish danio rerio. Tricaine ttbs western blot color development buffer. Native to south asia, it is a popular aquarium fish, frequently sold under the trade name zebra danio and thus often called a tropical fish although both tropical and subtropical. In the past two decades, zebrafish has become one of the preferred in vivo model organisms for studying diverse processes like embryogenesis 8, 9, autophagy, behaviour 11, 12 and sleep. Several factors have made the zebrafish an attractive vertebrate model for the study of motor development. A procedure for bleaching embryos can be found in the zebrafish book westerfield, 2000. The zebrafish is named for its five uniform, horizontal blue stripes on the side of its body, all of which extend from the gill cover back to the tail. Then, i apply tricaine 178 mgl for 60 seconds and view the larva under the microscope. Enhancing regeneration after acute kidney injury by. The name zebrafish comes from the horizontal blue stripes on each side of their bodies.
Capturing tissue repair in zebrafish larvae with timelapse. The zebrafish book a guide for the laboratory use of zebrafish danio brachydanio rerio by monte westerfield, institute of neuroscience, university of oregon. In the wild they inhabit clear, shallow slowmoving bodies of water with silt and wellvegetated bottoms. Each tank contained five green and five blue glass marbles to increase. Return the zebrafish to the swimming water to recover for 10 min. Kinesin family member 6 kif6 is necessary for spine. With a genome that has been sequenced and many molecular protocols in place, it has become. Histology and cytology both wildtype and spinal deformed sd zebrafish, were euthanized using a lethal dose of ms222 tricaine, fixed in. Feel free to add new protocols to the appropriate section or add comments to any existing protocol. Zebrafish larvae were injected with a single dose of gentamicin as previously described cianciolo cosentino et al.
Danio rerio are among the smallest members of this family, with adults measuring 30 40 millimeters, or about 1. For bone and cartilage staining, zebrafish at various time points were euthanized with tricaine methanesulfonate and fixed in 4% paraformaldehydephosphate buffered saline pfapbs. Current methods to visualize mineralized tissue in zebrafish rely on. However, it has been shown that tricaine can have effects on zebrafish cardiac function, and hence may bias the. Native to south asia, it is a popular aquarium fish, frequently sold under the trade name zebra danio and thus often called a tropical fish although both tropical and subtropical the zebrafish is an important and widely used vertebrate model organism in scientific research, for. Zebrafish danio rerio is a fresh water fish that inhabits rivers in india, pakistan and other places in asia. This research investigated the protective effect of lactobacillus plantarum against alcoholinduced liver injury and the regulatory mechanism of keapnrf2are signal pathway in zebrafish. Xenotransplantation of human glioblastoma in zebrafish. Add 2 ml of methylene blue stock solution, containing 0. A guide for the laboratory use of zebrafish danio brachydanio rerio. Two fish from the control group and one fish from the tricaine group had to be excluded due to methodological problems, leaving eight fish in the tricaine group and seven fish in the control group.
However, i think my middle schoolers might enjoy this book so i think ill still purchase it for my collection. The correct evaluation of mineralization is fundamental for the study of skeletal development, maintenance, and regeneration. Preparation of medium for zebrafish embryios e3 medium. Louis, mo, usa solution containing 2 gl nahco 3 wako pure chemical industries, ltd.
Zebrafish of 48 hpf were removed from the chorion, anesthetized with 200 mgl tricaine sigmaaldrich and mounted in agarose 0. Designed as a part of a larger commitment to the childrens hospital boston, the book stresses how individuals can make a difference locally and globally. Tricaine mesylate, finquel argent laboratories, red mond, w ashington, and tricaines w estern chemical, inc. However, i think my middle schoolers might enjoy this book so i. First, forward genetics can be applied to zebrafish to identify genes that are essential for proper behaviors granato et al. I anaesthetize larvae using 200mgl tricaine and make a 0. The inhibitory effect of tricaine on heart rate is reversed after 72 h. Methylene blue water 500 ml system water added with a small drop of concentrated methylene blue solution adult female fish keep female fish separate from male fish a week before egg collection. Capturing tissue repair in zebrafish larvae with time. Each dish was wrapped in parafilm then placed atop a 3inch spacer to achieve a dose rate of 978 cgymin in a 7 cesium source irradiator. Zebra fish definition of zebra fish by merriamwebster. Major insights into cardiac developmental processes have been gained by studies of embryonic zebrafish.
An earlier staging series for zebrafish, although less complete than the present one, fairly accurately por trays the first third or 1st day of embryonic develop. A paperback version of the fifth edition, called the zebrafish book. Anesthetize the fish by bath application of tricaine. However, it has been shown that tricaine can have effects on zebrafish cardiac function, and. Ab zebrafish embryos at 48 hpf were dechorionated, anesthetized with 0. Zebrafish international resource centers pathology services. Zebrafish are popular models for tissue regeneration because fish have the ability to regenerate numerous organs and body parts, including fins, 9 spinal cord tissue, 1 and heart 11 muscle. Images were taken just prior to amputation of the tail, immediately after the amputation, then every 3 days. Ct techniques, that are ultimately limited in resolution. Zebrafish definition of zebrafish by the free dictionary. A guide for the laboratory use of zebrafish danio rerio 4th ed. The same characteristics that make zebrafish a powerful system for basic research also make them an excellent tool for teaching. Zebrafish nacre strain were bred and raised according to established protocols. Ucsd guidelines for the preparation, storage and use of.
The tricaine dose was based on the recommendation in the zebrafish book. Zebrafish also seem to have the ability to regenerate photoreceptors and retinal neurons following injury. Fish were anesthetized with lowdose tricaine ms222 for phenotype analysis. The story follows a teenagers experience forming a band in her new school. Protocols are organized into sections corresponding to the chapters of the zebrafish book, 5th edition 4th edition online. It comes in a powdered form and should be stored at room temperature as a powder. Tricaine mesylate, finquel argent laboratories, red mond, w ashington, and tricaine s w estern chemical, inc. Today, the two web sites still appear very similar, because they are closely related in their mission to serve the research community.
Commonlyused tricaine doses in zebrafish anesthesia range from 150 to 200 mgl. Protective effect of lactobacillus plantarum on alcoholic. 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. An adult zebrafish was anesthetized with tricaine and had a tail clip taken for. Xenotransplantation of human glioblastoma in zebrafish larvae. Euthanatize fish with an overdose of ms222 tricaine methane sulfonate. Positioning the zebrafish to expose the ventral aspect, inject 50.
In its larval stages it is transparent and as it matures to an adult it develops stripes that run along the length of the body and look blue in colour. Behavioural effects of the commonly used fish anaesthetic. The celltrace far red labeled gbm cells were loaded into the glass needle at a density of about 3000cellsml. Highfat diet impairs cognitive function of zebrafish. All zebrafish used for experiments were wildtype, ab strain westerfield, 2000.
All procedures were approved by the uab institutional animal. This is where zebrafish researchers can share experimental protocols and tips with the rest of the research community. Laser ablation of the zebrafish pronephros to study renal. Includes antibodies submitted by the community and imported from zfin. Traditionally, cartilage is stained by alcian blue using acidic.
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