Today, it has emerged as a favored vertebrate organism for the research in science of reproduction. Some sites, such as footpad injection of freunds complete adjuvant, intrasplenic injection and intra lymph node injection are unacceptable nowadays ccac, 2002, and should. However, this method makes it difficult to know how much reagent is absorbed or taken up per fish. Procedure summary and goal describes procedure for the administration of fluids or compounds into the abdominal cavity. The zebrafish were anesthetized using the 2step anesthetic protocol detailed above. In contrast to mammals, the spinal cord of adult zebrafish has the capacity to reinitiate generation of motor neurons after a lesion. Anesthetics and analgesics, therapeutic agents, and test compounds must frequently be administered to animal subjects by injection. The zebrafish book university of oregon press, eugene 1995. Glp 106 ip injection of rats and mice university of newcastle. We demonstrate intraperitoneal injection into adult zebrafish. If there is negative pressure, proceed with the injection. For their physical characteristics, including small size, easiness to grow, and robustness of the species, zebrafish danio rerio is rapidly becoming a popular model in biobehavioral studies. The fate of intraperitoneally injected carbon particles in.
There is a significant growth in amount numbers of scientific literature pertaining to research discoveries in reproductive sciences in zebrafish. Once anesthetized, intraperitoneal injections were performed using a beveled, 26sguaged hamilton syringe hamilton. The laboratory mouse handbook of experimental animals. I agree to abide by the risk controls outlined in this assessment. Thus, retroorbital injection may greatly facilitate zebrafish research requiring chemical or chemical delivery. While the most popular drug administration technique in zebrafish research is by immersion, systemic intraperitoneal injection is another effective alternative. Longterm drug administration in the adult zebrafish using oral. Intramuscular im is my first choice for injection and is used if the koi has ulcers that have broken through to the body cavity. It is more often applied to animals than to humans.
I have read and understood the risk assessment for glp 106 intraperitoneal injection of rats and mice. I need to perform intraperitoneal injections of a highly hydrophobic compound in adult zebrafish. Some experimental questions, particularly those related to metabolic studies, may be better addressed by delivering a defined quantity to each fish, based on weight. Soaking zebrafish embryos in ptyellow or intraperitoneal injection into adults results in nontoxic in vivo fluorescent labeling of the renal proximal tubules, the major site of blood filtrate reabsorption and a common target of injury in acute kidney injury. This practice is taken from the fish veterinary literature e. Zebrafish are also easy to manipulate for researchers who. They laugh harder when i say, of course i put them to sleep first.
Intraperitoneal injection in mice research at penn state. Steroidogenic acute regulatory protein and luteinizing. The intramuscular administration is not recommended, as the muscle of the mouse is too small. Dec 07, 2009 further, since ro injection is faster and easier than intracardiac injection techniques in the adult fish, another application of ro injection may include adult chemical genetic screening.
Pdf intraperitoneal injection into adult zebrafish researchgate. Success includes a healthy fish patient as well as a good experimental outcome. Here we demonstrate that adult zebrafish, which show functional spinal cord regeneration, are capable of motor neuron regeneration. This procedure is adapted from veterinary methods used for larger. Ensure that all equipment is available prior to starting the procedure. During the first day, carbon was phagocytized by macrophages situated in the spleen within the ellipsoids. Key wordszebrafishintraperitoneal injectiondrug exposuredrug administration methodanxiety. Intraperitoneal injection of 2 mgkg in rat neonates, 15 mgkg in adult rats and 20 mgkg in mice, 6 mgkg in zebrafish. Although characterized in detail in their structure and activity, the temporal and spatial expression of amps during vertebrate embryogenesis is still poorly understood. E2f8 promotes hepatic steatosis through fabp3 expression. Intraperitoneal injection of rats and mice animal house personnel. Fullerenes c 60 are nanomaterials with unique physicochemical and biological properties that are important for the development of many technological applications.
Zebrafish danio rerio as a model to visualize infection. Intraperitoneal injection into adult zebrafish ncbi nih. The small molecule probe ptyellow labels the renal. The system utilizes a gradual cooling method for the anesthesia and maintains the fish in a stable anesthetic plane, as well as stabilizes the animal so that intraperitoneal injections can be consistently performed. Disease models and chemical screens, volume 8 4th edition. A novel, lowcost anesthesia and injection system for zebrafish. Intraperitoneal injection is widely used in rodents as a route of drug administration. Injection often referred to as a shot in us english, or a jab in uk english is the act of putting a liquid, especially a drug, into a persons body using a needle usually a hypodermic needle and a syringe. Development of a novel zebrafish model for type 2 diabetes. Scientists developed a new lithiumsulphur battery with a capacity five times higher than that of lithiumion batteries, which maintains an efficiency of 99% for more than 200 cycles, and may keep a smartphone charged for five days.
Intraperitoneal dye injection method for visualizing the. Intrathoracic injection for the study of adult zebrafish. The lymphoid organs of rosy barb barbus conchonius and carp cyprinus carpio were investigated for their phagocytic uptake of carbon, after its intraperitoneal injection. We report the development of a small fluorescent molecule, bdnca3d2, herein referred to as ptyellow. I drank celery juice for 7 days and this is what happened no juicer required. Intraperitoneal injection into adult zebrafish europe.
Injection is into the abdominal cavity, posterior to the pelvic girdle. Intraperitoneally applied drug forms are supposed to be located in the peritoneal cavity. However, formatting rules can vary widely between applications and fields of interest or study. For each step there are best practices that can ensure success.
It was first detected in the lymphoid organs at 30 min after injection. By injecting sequences containing recognition sites for dna modifying enzymes, scientists can efficiently generate transgenic fish with modified genomes, meaning that foreign genes will be passed on to future generations. An affordable intraperitoneal injection setup for juvenile and adult. Zebra fish microinjection instruments for zebrafish research oocytes, embryos, larvae and adults world precision instruments wpi offers a broad range of research instruments for zebrafish danio rerio research, from the time the genetics of the subject are modified through the point where the physiological effects of the change are monitored. Within the cavity of the peritoneum intraperitoneal. Tricaine did not show sig nificant alterations in behavior in the 6min novel tank test fig. This enables accurate adjustment of the infection dose. With big fish i usually inject this type of chemicals dissolved in oil, but in zebrafish, if i.
Intraperitoneal injection in the mouse procedures with care. Following a described protocol for zebrafish intraperitoneal injection 78. Intraperitoneal injection or ip injection is the injection of a substance into the peritoneum body cavity. However, experimental infections are usually introduced by intraperitoneal injection in anaesthetized fish. Their expression can be constitutive or inducible by infectious challenge. The aim of this study was to evaluate the differences in histopathological structure and activities and transcription levels of antioxidant enzymes cat, sod, gpx, gst in the liver of zebrafish i. Zebrafish pd model induced by 1methyl4phenyl1,2,3,6tetrahydropyridine showed a significant level of decrease in the movement with erratic swimming pattern and increased freezing bouts. However, current ip equipment can be either costly, inaccurate, or unsafe for zebrafish. Pull back on the plunger to ensure negative pressure prior to injecting.
A guide for the laboratory use of zebrafish danio rerio, 4th edn. Conversely, intraperitoneal serotonin injections doubled numbers of new motor neurons and proliferating pmnlike progenitors caudal to the lesion. First, adult zebrafish received a single 30 mgkg intraperitoneal injection of fluorescentlytagged gentamicin gttr to determine the distribution of gentamicin within inner ear sensory epithelia. Injection of 10 3 bacteria per fish resulted in approximately 50% mortality by day 4 post injection. Intraperitoneal injection in the rat procedures with care. Pdf intraperitoneal injection into adult zebrafish semantic scholar. Seizures induced resemble human temporal lobe epilepsy. Intraperoteneal ip, intramuscular im, and intravenous iv. A convenient method for chemically treating zebrafish is to introduce the reagent into the tank water, where it will be taken up by the fish. An affordable intraperitoneal injection setup for juvenile. Even though technologies involving nanomicroparticles have great potential, it is crucial to determine possible toxicity of these technological products before extensive use. The disappearance of drugs from the peritoneal cavity upon injection is because of diffusion into the surrounding tissues.
Aug 30, 2010 some experimental questions, particularly those related to metabolic studies, may be better addressed by delivering a defined quantity to each fish, based on weight. Here we present a method for intraperitoneal ip injection into adult zebrafish. The fish may take 10 to 15 minutes to revive in the oxygenated tank, depending on the strength of the anesthetic and the particular fishes response. Antimicrobial peptides amps are ancestral components in the evolution of immunity from protozoans to metazoans. Intraperitoneal injection of trypan blue is useful for temporal observations of the lymphatic ducts, which are essential for large. Prince 1 1 department of organismal biology and anatomy, the university of chicago, 2 committee on molecular metabolism and nutrition, the university of chicago, 3 department of medicine, the university of chicago.
Intraperitoneal ip injections should be made into the animals right abdominal quadrant so as to avoid penetrating the cecum on the left side. Intraperitoneal injection is usually administered just to the right of the midline at the level of the umbilicus. Intraperitoneal injections of oil in adult zebrafish. Zebrafish a model system biology essay freebooksummary.
Intraperitoneal injection into adult zebrafish core. Intraperitoneal injection definition of intraperitoneal. Compared to intraperitoneal ip injections, the main advantage of intrathoracic injections is the focal administration of the tested factors on the target organ. In this study, we designed and developed a novel lowcost system for anesthetizing and injecting adult zebrafish. Here we use zebrafish as a model to visualize and compare the development of infection of vibrio anguillarum on and in the fish following injection or bathing. Currently, intraperitoneal ip injection of dluciferin is the preferred method of providing substrate for bioluminescence imaging bli. Here we introduce a novel anesthetic combination of ms222 and.
To encoding the acutephase proteins the aeromonas salmonicida, a gm negative bacteria and staphylococcus aureus induced the cistrons in zebra fish. A significantly lower mortality was achieved in all vaccinated groupsboth the prime vaccinated and boosted groupscompared with nonvaccinated controls. Intraperitoneal injection into adult zebrafish jove this protocol presents a method for intraperitoneal ip injection into adult zebrafish. Intraperitoneal injection into adult zebrafish mary d. Any medication injected into the peritoneal cavity in a fish of this condition. Motor neuron regeneration in adult zebrafish journal of. Intraperitoneal injection in the mouse although widely used as a means of administering substances, particularly injectable anaesthetics, this is an inherently unreliable technique, since inadvertent injection of some material into the gut, abdominal fat and subcutaneous tissues is a relatively frequent occurrence. Zebrafish embryos are infected at between 28 or 30 hpf by microinjection. Alternative sites as described for other small mammals may also be applicable. Zebrafish, intraperitoneal injection, drug exposure, drug. This procedure is adapted from veterinary methods used for larger fish. Induces limbic seizures characterized by a seizure scale devised by racine. Here we show that genes involved in motor neuron development, i. Intraperitoneal ip injection in rats and mice sop page 4 of 6 mouse rat 6.
The cross reactivity of daily intraperitoneal injection of 0. Zebrafish were overfed a commercially available fish food using an automated feeding system. Training should be undertaken in general laboratory safety. Regeneration of spinalintrinsic serotonergic interneurons was unaltered by these manipulations.
Intraperitoneal ip injections are an effective and reproducible route of drug administration. Injection is a technique for delivering drugs by parenteral administration, that is, administration via a route other than through the digestive tract. Citeseerx chapter 14 intraperitoneal injection as a. From here on we followed the zebrafish book, 4th edition, but kept the head of each embryo for genotyping. The delivery of molecules directly into the pericardium is a suitable strategy for studies of cardiac preconditioning and. Vectashield mounting medium with dapi was used to visualize the spreads. The fish were challenged by intraperitoneal injection of 9. Longterm drug administration in the adult zebrafish using. We describe a simple, lowcost ip setup, which can be easily assembled from inexpensive and readily available parts, and which provides a safe, reproducible, and. The truth is that every koi hobbyist will one day need to inject their fish either with antibiotics or with a vaccine. Intraperitoneal ip injection of zebrafish is considered an effective and reproducible route of administration to study various biological and. Hart and siddharth gaikwad and eli utterback and john dileo and allan v. Abstract zebrafish behavioral phenotypes are often evaluated in response to pharmacological modulation by various psychotropic drugs. Intraperitoneal injection of the nephrotoxin gentamicin results in acute kidney injury and a regeneration response characterized by synchronized production of new nephrons reimschuessel, 2001.
A novel, lowcost anesthesia and injection system for. Aminoglycosideinduced hair cell death of inner ear organs. A novel, lowcost anesthesia and injection system for zebrafish researchers. Intraperitoneal injection carries the risk of causing internal injury or of misplaced injection into a blood vessel or an organ, particularly the large thinwalled cecum or urinary bladder. The laboratory rabbit, guinea pig, hamster, and other rodents, 2012. Intraperitoneal injection intraperitoneal injection demons. An important step in this process is the method of drug administration. Injection of laboratory animals parenteral injections the ability to administer materials by injection is essential for most experimental studies employing laboratory animals. People often laugh when i tell them that i inject my fish. Loss of the homologous recombination gene rad51 leads to. Feb 04, 2015 zebrafish intraperitoneal injection ayca arslan ergul.
This method is useful for drugs that are difficult to dissolve in water, or which require a better control over the amount of drug delivered to an individual animal. Zebrafish patient avatars in cancer biology and precision. We also conducted in vitro overexpression in human cells. Rojo et al, non long ago explained that due to the response of l. In general, it is preferred when large amounts of blood replacement fluids are needed or when low blood pressure or other problems prevent the use of a suitable blood vessel for intravenous injection. The zebrafish international resource center provides a central repository for wildtype and mutant strains of zebrafish danio rerio and for materials and information about zebrafish research. Hence, serotonin selectively promotes the development and adult regeneration of motor neurons in. Video article intraperitoneal injection into adult zebrafish, year. Aug 20, 2008 the mammalian spinal cord does not regenerate motor neurons that are lost as a result of injury or disease. Jul 08, 2012 the location i give the ip injection was instructed by my mentor veterinarian. Wnt signaling mediates new nephron formation during. This will help prevent hypoglycemic seizures that can occur when a lamb is warmed up. Intraperitoneal exposure to nanomicroparticles of fullerene. Intramuscular im, intraperitoneal ip, and intravenous iv.
Intraperitoneal injection into adult zebrafish protocol. Intraperitoneal ip injection in rats and mice sop page 2 of 6 70% isopropyl alcohol to disinfect top of multi. World precision instruments zebrafish microinjection. Anesthetized fish were placed dorsal side up on a damp sponge and stabilized with one hand. Intraperitoneal injection of glucose if a lamb greater than 5 hours old is suffering from hypothermia, it is essential to provide it with an energy source prior to warming. Hepatic positive and negative antioxidant responses in. Glucose delivery to adult zebrafish using intraperitoneal injection. Serotonin promotes development and regeneration of spinal. Zebra fish models for diabetes linkedin slideshare.
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