This work was supported by NIH grant R01 HD050901 to MCM and American Cancer Society Postdoctoral Fellowship #PF-05-041-01-DDC to TG.

<ol>
<li>A male zebrafish will be dissected first, followed by a female fish. Before beginning the dissection, anesthetize a fish in 0.2% Tricaine and then euthanize it by incubation in ice water for 15 minutes.
</li>
<li>Begin by lightly patting the fish dry on a paper towel and placing it on a dissecting mat. Externally, zebrafish have single dorsal, caudal and anal fins and paired pectoral and pelvic fins (Figure 1). 
 <img src="/files/ftp_upload/1717/1717fig1.jpg" alt="Zebrafish anatomy diagram; labeled fins: dorsal, caudal, pectoral, pelvic, anal; educational use." data-altupdatedat="1747911167000" data-alt="Figure 1"> Figure 1. Adult male fish with fins labeled. </li>
<li>Pin the fish to the dissecting mat through the fleshy part of the tail and the ventral part of the eye socket.</li>
<li>Snip the skin on the belly of the fish just anterior to the anal fin. Cut the skin and underlying muscle along the belly from the anal fin to the operculum (the hard covering over the gill) (Figure 2, step 1). <img src="/files/ftp_upload/1717/1717fig2.jpg" alt="Zebrafish anatomy diagram with numbered regions for developmental biology research or genetic studies." data-altupdatedat="1747911167000" data-alt="Figure 2"> Figure 2. Steps in removing skin and body wall muscle to expose internal organs. </li>
<li>Next, remove the operculum and the pectoral fin, including the pectoral girdle (the thick, bony region at the base of the fin) (Figure 2, step 2). Cut the skin and underlying muscle beginning from above the now exposed gill posteriorly along the side of the fish and then down to the anal fin (Figure 2, step 3).</li>
<li>Carefully remove the skin and underlying muscle from the side of the fish. Many of the internal organs are now visible (Figure 3). 
<img src="/files/ftp_upload/1717/1717fig3.jpg" alt="Zebrafish anatomy diagram highlighting heart, liver, intestine, testis, and swim bladder." data-altupdatedat="1747911167000" data-alt="Figure 3"> Figure 3. Adult male fish with body wall and muscle removed allowing the visualization of the internal organs. </li>
<li>The testes are long, white, paired organs that are attached to the dorsal body wall. Remove a testis and place it in a dish of PBS. Examine the testis with reflected light to visualize the seminiferous tubules (Figure 4), which contain cysts with various stages of developing germ cells from spermatagonia to spermatids (Leal et al., 2009). <img src="/files/ftp_upload/1717/1717fig4.jpg" alt="Microscope image of plant root cells showing cellular structure and organization for biological study." data-altupdatedat="1747911167000" data-alt="Figure 4"> Figure 4. Dissection of a testis. The white roundish structures are the seminiferous tubules. </li>
<li>Next, remove the gastrointestinal system from the body cavity of the fish. The liver can be identified by its large size, lobed morphology, tannish color, and extensive vascularization. The gallbladder, a green translucent fluid-filled sac, and the spleen, which appears bright red, are found within the viscera.</li>
<li>Separate the intestine from the rest of the organs and stretch it out. The anterior, mid, and posterior regions of the intestine are defined by the height of the epithelial folds (Wallace et al., 2005).</li>
<li>Place a piece of the intestine in PBS and observe the epithelial folds using transmitted light.</li>
<li>Next, examine the swim bladder. The swim bladder consists of a posterior chamber, which is connected to the esophagus via the pneumatic duct, and an anterior chamber, which is connected to the inner ear through the Weberian apparatus (Finney et al., 2006).</li>
<li>Remove and discard the swim bladder. Unpin the fish and re-pin it ventral side up to dissect the kidney, which is located along the dorsal body wall. The kidney is a translucent pink structure associated with the dorsal aorta and pigmented cells. The kidney is divided into head, body, and tail regions (Figure 5). Dissect out a piece of the kidney and place it in PBS. Tease apart the kidney tissue to reveal the renal tubules. <img src="/files/ftp_upload/1717/1717fig5.jpg" alt="Earthworm anatomy diagram showing head, body, tail labeled for educational study." data-altupdatedat="1747911167000" data-alt="Figure 5"> Figure 5. Location of head, body, and tail kidney along the dorsal body wall. </li>
<li>Dissect a female fish. As described previously, euthanize the fish in ice water and pat it dry before pinning it to the dissecting mat. Remove the skin from the side of the fish as previously demonstrated. The ovary is a bilobed structure that is suspended in the body cavity by a vascularized mesovarium.</li>
<li>Remove one lobe of the ovary, place it in PBS, and examine it with transmitted light. The oocytes can be teased apart using fine needles and then staged (Figure 6, Selman et al., 1993). Stage I oocytes are approximately 10 - 150 microns in size and translucent. Stage II oocytes are about 150 - 350 microns in size and defined by the appearance of cortical granules. Stage III oocytes are 350 - 750 microns in size and are opaque due to the accumulation of yolk. Mature stage IV and V oocytes are translucent and generally not found in the ovaries of females that have recently mated. 
<img src="/files/ftp_upload/1717/1717fig6.jpg" alt="Cell growth under microscope, labeled I-III, biology experiment, cellular development analysis, diagram." data-altupdatedat="1747911167000" data-alt="Figure 6"> Figure 6. Live stage I, II, and III oocytes observed under a dissecting microscope with transmitted light. </li>
<li>Next, dissect the heart from the fish. The heart is located posterior and ventral to the gill. Begin by cutting out the heart and all of the surrounding tissue and placing it in PBS. Carefully dissect away the tissue surrounding the heart, being careful not to damage the delicate atrium.</li>
<li>Place the dissected heart into Ringer's solution to observe the heart beat. Identify the atrium, ventricle, and bulbus arteriosus (Figure 7, Hu et al., 2001). <img src="/files/ftp_upload/1717/1717fig7.jpg" alt="Heart anatomy diagram with bulbus arteriosus, atrium, ventricle labeled; cardiac structure study." data-altupdatedat="1747911167000" data-alt="Figure 7"> Figure 7. Dissected adult heart. </li>
<li>Complete the dissection by removing the brain from the fish. Begin by unpinning the fish and removing the head with a razor blade. Remove as much soft tissue as possible from the ventral side of the skull with forceps. Remove the eyes using small spring scissors. Break open the skull and remove the bone from the ventral side of the brain. Now place the head in a dish of PBS and remove the skin and skull bones from the dorsal side of the brain.</li>
<li>Identify the olfactory bulbs, telencephalon, habenula, optic tectum, cerebellum and medulla (Figure 8, Wullimann et al., 1996; Schilling, 2002). <img src="/files/ftp_upload/1717/1717fig8.jpg" alt="Zebrafish brain anatomy diagram with labeled sections: olfactory bulb, telencephalon, cerebellum." data-altupdatedat="1747911167000" data-alt="Figure 8"> Figure 8. Dissected adult zebrafish brain. Dorsal view, anterior to top. </li>
</ol>

<ol>
	<li>Finney, J. L., Robertson, G. N., McGee, C. A., Smith, F. M., Croll, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+Autonomic+Innervation+of+the+Swim+Bladder+in+the+Zebrafish+(Danio+rerio).">Structure and Autonomic Innervation of the Swim Bladder in the Zebrafish (Danio rerio).</a> J Comp Neurol. 495, 587-606 (2006).</li><li>Hu, N., Yost, H. J., Clark, E. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+morphology+and+blood+pressure+in+the+adult+zebrafish.">Cardiac morphology and blood pressure in the adult zebrafish.</a> Anat Rec. 264, 1-12 (2001).</li><li>Leal, M. C., Cardoso, E. R., N&#243;brega, R. H., Batlouni, S. R., Bogerd, J., Fran&#231;a, L. R., Schulz, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+and+stereological+evaluation+of+zebrafish+(Danio+rerio)+spermatogenesis+with+an+emphasis+on+spermatogonial+generations.">Histological and stereological evaluation of zebrafish (Danio rerio) spermatogenesis with an emphasis on spermatogonial generations.</a> Biol Reprod. 81, 177-187 (2009).</li><li>Schilling, T. F., Nusslein-Volhard, C., Dahm, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+morphology+of+larval+and+adult+zebrafish.">The morphology of larval and adult zebrafish.</a> Zebrafish: A Practical Approach (The Practical Approach Series). , (2002).</li><li>Selman, K., Wallace, R., Sarka, A., Qi, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stages+of+oocyte+development+in+the+Zebrafish,+Brachydanio+rerio.">Stages of oocyte development in the Zebrafish, Brachydanio rerio.</a> J. Morphol. 218, 203-224 (1993).</li><li>Wallace, K. N., Akhter, S., Smith, E. M., Lorent, K., Pack, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intestinal+growth+and+differentiation+in+zebrafish.">Intestinal growth and differentiation in zebrafish.</a> Mech Dev. 122, 157-173 (2005).</li><li>Wullimann, M. F., Rupp, B., Reichert, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Neuroanatomy of the zebrafish brain : a topological atlas. , (1996).</li></ol>

Experiments on animals were performed in accordance with the guidelines and regulations set forth by the University of Pennsylvania Institutional Animal Care and Use Committee.

<table><tbody><tr><td colspan="4">Reagents:</td></tr><tr><td colspan="4">&lt;strong&gt;&lt;em&gt;0.2% Tricaine (ethyl 3-aminobenzoate)&lt;/em&gt;&lt;/strong&gt;&lt;br/&gt;200 mg tricaine powder&lt;br/&gt;97.9 ml DD water&lt;br/&gt;~1 ml 1 M Tris (pH 9)&lt;br/&gt;Adjust pH to 7.0</td></tr><tr><td colspan="4">&lt;strong&gt;&lt;em&gt;Phosphate buffered saline (PBS)&lt;/em&gt;&lt;/strong&gt;&lt;br/&gt;4.0 g NaCl&lt;br/&gt;0.1 g KCl&lt;br/&gt;100 ml 0.1 M PO&lt;sub&gt;4&lt;/sub&gt; Buffer, pH 7.3&lt;br/&gt;150 ml dH&lt;sub&gt;2&lt;/sub&gt;O</td></tr><tr><td colspan="4">&lt;strong&gt;&lt;em&gt;Ringer&amp;rsquo;s solution&lt;/em&gt;&lt;/strong&gt;&lt;br/&gt;6.7g NaCl&lt;br/&gt;0.2g KCl&lt;br/&gt;0.2g CaCl&lt;sub&gt;2&lt;/sub&gt;&lt;br/&gt;1.2g Hepes&lt;br/&gt;1 L H&lt;sub&gt;2&lt;/sub&gt;O &lt;br/&gt;Adjust pH to 7.2</td></tr><tr><td>Dissecting dish with mat</td><td>Electron Microscopy Sciences</td><td>70540</td><td></td></tr><tr><td>Vannas spring scissors</td><td>Fine Science Tools</td><td>91500-09</td><td></td></tr></tbody></table>

dissection of organs from the adult zebrafish

Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease.  Although experimental analysis of the embryo and larva is extensive and the morphology has been well documented, descriptions of adult zebrafish anatomy and studies of development of the adult structures and organs, together with techniques for working with adults are lacking. The organs of the larva undergo significant changes in their overall structure, morphology, and anatomical location during the larval to adult transition.  Externally, the transparent larva develops its characteristic adult striped pigment pattern and paired pelvic fins, while internally, the organs undergo massive growth and remodeling.  In addition, the bipotential gonad primordium develops into either testis or ovary.  This protocol identifies many of the organs of the adult and demonstrates methods for dissection of the brain, gonads, gastrointestinal system, heart, and kidney of the adult zebrafish. The dissected organs can be used for in situ hybridization, immunohistochemistry, histology, RNA extraction, protein analysis, and other molecular techniques.  This protocol will assist in the broadening of studies in the zebrafish to include the remodeling of larval organs, the morphogenesis of organs specific to the adult and other investigations of the adult organ systems.

Watch this Scientific Journal Video about Dissection of Organs from the Adult Zebrafish at JoVE.com

Dissection of Organs from the Adult Zebrafish

This protocol describes a procedure for identifying and dissecting organs from the adult zebrafish.

Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease.  Although experimental ...

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104.6K Views.  University of Pennsylvania-School of Medicine. This protocol describes a procedure for identifying and dissecting organs from the adult zebrafish.

Video: Dissection of Organs from the Adult Zebrafish

Large-scale Zebrafish Embryonic Heart Dissection for Transcriptional Analysis

The zebrafish embryonic heart is composed of only a few hundred cells, representing only a small fraction of the entire embryo. Therefore, to prevent the cardiac transcriptome from being masked by the global embryonic transcriptome, it is necessary to collect sufficient numbers of hearts for further analyses. Furthermore, as zebrafish cardiac development proceeds rapidly, heart collection and RNA extraction methods need to be quick in order to ensure homogeneity of the samples. Here, we present a rapid manual dissection protocol for collecting functional/beating hearts from zebrafish embryos. This is an essential prerequisite for subsequent cardiac-specific RNA extraction to determine cardiac-specific gene expression levels by transcriptome analyses, such as quantitative real-time polymerase chain reaction (RT-qPCR). The method is based on differential adhesive properties of the zebrafish embryonic heart compared with other tissues; this allows for the rapid physical separation of cardiac from extracardiac tissue by a combination of fluidic shear force disruption, stepwise filtration and manual collection of transgenic fluorescently labeled hearts.

To analyse cardiac gene expression profiles during zebrafish heart development, total RNA has to be extracted from isolated hearts. Here, we present a protocol for collecting functional/beating hearts by rapid manual dissection from zebrafish embryos to obtain cardiac-specific mRNA.

The zebrafish embryonic heart is composed of only a few hundred cells, representing only a small fraction of the entire embryo. Therefore, to prevent the ...

Developmental Biology

Dissection of Larval Zebrafish Gonadal Tissue

Although wild zebrafish possess a ZZ/ZW sex-determination system, domesticated zebrafish have lost the sex chromosome. They utilize a polygenic sex determination system, where several genes distributed throughout the genome collectively determine the sex identities of individual fish. Currently, the genes involved in regulating gonad development and how they work remain elusive. Normally, isolating gonadal tissue is the first step to examine the sex developmental processes. Here, we present a procedure to isolate gonadal tissue from 17 dpf (days post fertilization) and 25 dpf zebrafish larvae. The isolated gonadal tissue may be subsequently examined by morphology and gene expression profiling.

Here, we present a protocol for isolating gonadal tissue of larval zebrafish, which will facilitate investigations of zebrafish sex differentiation and maintenance.

Although wild zebrafish possess a ZZ/ZW sex-determination system, domesticated zebrafish have lost the sex chromosome. They utilize a polygenic sex ...

Dissection of the Adult Zebrafish Kidney

Researchers working in the burgeoning field of adult stem cell biology seek to understand the signals that regulate the behavior and function of stem cells during normal homeostasis and disease states. The understanding of adult stem cells has broad reaching implications for the future of regenerative medicine1. For example, better knowledge about adult stem cell biology can facilitate the design of therapeutic strategies in which organs are triggered to heal themselves or even the creation of methods for growing organs in vitro that can be transplanted into humans1. The zebrafish has become a powerful animal model for the study of vertebrate cell biology2. There has been extensive documentation and analysis of embryonic development in the zebrafish3. Only recently have scientists sought to document adult anatomy and surgical dissection techniques4, as there has been a progressive movement within the zebrafish community to broaden the applications of this research organism to adult studies. For example, there are expanding interests in using zebrafish to investigate the biology of adult stem cell populations and make sophisticated adult models of diseases such as cancer5. Historically, isolation of the zebrafish adult kidney has been instrumental for studying hematopoiesis, as the kidney is the anatomical location of blood cell production in fish6,7. The kidney is composed of nephron functional units found in arborized arrangements, surrounded by hematopoietic tissue that is dispersed throughout the intervening spaces. The hematopoietic component consists of hematopoietic stem cells (HSCs) and their progeny that inhabit the kidney until they terminally differentiate8. In addition, it is now appreciated that a group of renal stem/progenitor cells (RPCs) also inhabit the zebrafish kidney organ and enable both kidney regeneration and growth, as observed in other fish species9-11. In light of this new discovery, the zebrafish kidney is one organ that houses the location of two exciting opportunities for adult stem cell biology studies. It is clear that many outstanding questions could be well served with this experimental system. To encourage expansion of this field, it is beneficial to document detailed methods of visualizing and then isolating the adult zebrafish kidney organ. This protocol details our procedure for dissection of the adult kidney from both unfixed and fixed animals.
Dissection of the kidney organ can be used to isolate and characterize hematopoietic and renal stem cells and their offspring using established techniques such as histology, fluorescence activated cell sorting (FACS)11,12, expression profiling13,14, and transplantation11,15. 
We hope that dissemination of this protocol will provide researchers with the knowledge to implement broader use of zebrafish studies that ultimately can be translated for human application.

The zebrafish kidney is home to both renal and hematopoietic adult stem/progenitor cells, and represents an outstanding opportunity to study these cell types and their progeny in a vertebrate model organism. Here, we demonstrate a detailed dissection procedure that enables the researcher to identify and surgically remove the adult zebrafish kidney, which can be used for applications such as cell isolation, transplantation, and expression studies of kidney and/or blood cell populations.

Researchers working in the burgeoning field of adult stem cell biology seek to understand the signals that regulate the behavior and function of stem ...

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work examining post-embryonic through adult cardiac development has been limited7-10. Examining the changing morphology of the maturing and aging heart are restricted by the lack of techniques available for staging and isolating juvenile and adult hearts. In order to analyze heart development over the fish's lifespan, we dissect zebrafish hearts at numerous stages and photograph them for further analysis11. The morphological features of the heart can easily be quantified and individual hearts can be further analyzed by a host of standard methods. Zebrafish grow at variable rates and maturation correlates better with fish size than age, thus, post-fixation, we photograph and measure fish length as a gauge of fish maturation. This protocol explains two distinct, size dependent dissection techniques for zebrafish, ranging from larvae 3.5mm standard length (SL) with hearts of 100&mu;m ventricle length (VL), to adults, with SL of 30mm and VL 1mm or larger. Larval and adult fish have quite distinct body and organ morphology. Larvae are not only significantly smaller, they have less pigment and each organ is visually very difficult to identify. For this reason, we use distinct dissection techniques.
We used pre-dissection fixation procedures, as we discovered that hearts dissected directly after euthanization have a more variable morphology, with very loose and balloon like atria compared with hearts removed following fixation. The fish fixed prior to dissection, retain in vivo morphology and chamber position (data not shown). In addition, for demonstration purposes, we take advantage of the heart (myocardial) specific GFP transgenic Tg(myl7:GFP)twu34 (12), which allows us to visualize the entire heart and is particularly useful at early stages in development when the cardiac morphology is less distinct from surrounding tissues. Dissection of the heart makes further analysis of the cell and molecular biology underlying heart development and maturation using in situ hybridization, immunohistochemistry, RNA extraction or other analytical methods easier in post-embryonic zebrafish. This protocol will provide a valuable technique for the study of cardiac development maturation and aging.

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

A clear, standardized method for dissection and isolation of the zebrafish heart at multiple developmental stages are described. Annotation and quantification techniques are also discussed.

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work ...

Analysis of Nephron Composition and Function in the Adult Zebrafish Kidney

The zebrafish model has emerged as a relevant system to study kidney development, regeneration and disease. Both the embryonic and adult zebrafish kidneys are composed of functional units known as nephrons, which are highly conserved with other vertebrates, including mammals. Research in zebrafish has recently demonstrated that two distinctive phenomena transpire after adult nephrons incur damage: first, there is robust regeneration within existing nephrons that replaces the destroyed tubule epithelial cells; second, entirely new nephrons are produced from renal progenitors in a process known as neonephrogenesis. In contrast, humans and other mammals seem to have only a limited ability for nephron epithelial regeneration. To date, the mechanisms responsible for these kidney regeneration phenomena remain poorly understood. Since adult zebrafish kidneys undergo both nephron epithelial regeneration and neonephrogenesis, they provide an outstanding experimental paradigm to study these events. Further, there is a wide range of genetic and pharmacological tools available in the zebrafish model that can be used to delineate the cellular and molecular mechanisms that regulate renal regeneration. One essential aspect of such research is the evaluation of nephron structure and function. This protocol describes a set of labeling techniques that can be used to gauge renal composition and test nephron functionality in the adult zebrafish kidney. Thus, these methods are widely applicable to the future phenotypic characterization of adult zebrafish kidney injury paradigms, which include but are not limited to, nephrotoxicant exposure regimes or genetic methods of targeted cell death such as the nitroreductase mediated cell ablation technique. Further, these methods could be used to study genetic perturbations in adult kidney formation and could also be applied to assess renal status during chronic disease modeling.

The zebrafish adult kidney is an excellent system for renal regeneration and disease studies. An essential aspect of such research is the assessment of nephron structure and function. This protocol describes several methodologies that can be implemented to assess nephron tubule composition and to evaluate renal reabsorption.

The zebrafish model has emerged as a relevant system to study kidney development, regeneration and disease. Both the embryonic and adult zebrafish kidneys ...

Recovery of Adult Zebrafish Hearts for High-throughput Applications

Use of the zebrafish model system for studying development, regeneration, and disease is expanding toward use of adult hearts for cell dissociation and purification of RNA, DNA, and proteins. All of these applications demand the rapid recovery of significant numbers of zebrafish hearts to avoid gene regulatory, metabolic, and other changes that begin after death. Adult zebrafish hearts are also required for studying heart structure for a variety of mutants and for studying heart regeneration. However, the traditional zebrafish heart dissection is slow and difficult and requires specialized tools, making large-scale dissection of adult zebrafish hearts tedious. Traditional methods also harbor the risk of damaging the heart during the dissection. Here, we describe a method for dissection of adult zebrafish hearts that is fast, reproducible, and preserves heart architecture. Furthermore, this method does not require specialized tools, is painless for the zebrafish, can be performed on fresh or fixed specimens, and can be performed on zebrafish as young as one month old. The approach described expands the use of adult zebrafish for cardiovascular research.

Use of zebrafish for cardiovascular research is expanding towards research on adult hearts. For these applications, quick and simple isolation of cardiac tissues is key to avoid post-mortem changes and to obtain an adequate number of samples. Here, we describe a fast and reproducible method for dissecting adult zebrafish hearts.

Use of the zebrafish model system for studying development, regeneration, and disease is expanding toward use of adult hearts for cell dissociation and ...

Partial Hepatectomy in Adult Zebrafish

Liver failure is one of the leading causes of death worldwide, and mortality from chronic liver disease is rising sharply in the United States. Healthy livers are capable of regenerating from toxic damage, but in advanced liver disease, the natural ability of the liver to regenerate is impaired. Zebrafish have emerged as a powerful experimental system for studying regeneration. They are an ideal model for studying liver regeneration from partial hepatectomy, a procedure with direct clinical relevance in which part of the liver is surgically removed, leaving the rest intact. There is no standard protocol for partial hepatectomy; previous studies using this model have used slightly different protocols and reported disparate results. Described here is an efficient, reproducible protocol for performing a partial hepatectomy in adult zebrafish. We use this technique to demonstrate that zebrafish are capable of epimorphic regeneration of the resected lobe. This protocol can be used to further interrogate the mechanisms required for liver regeneration in zebrafish.

This protocol describes the procedure for removing the ventral lobe of the liver in adult zebrafish to enable the study of liver regeneration.

Liver failure is one of the leading causes of death worldwide, and mortality from chronic liver disease is rising sharply in the United States. Healthy ...

Isolation of Cardiac and Vascular Smooth Muscle Cells from Adult, Juvenile, Larval and Embryonic Zebrafish for Electrophysiological Studies

Zebrafish have long been used as a model vertebrate organism in cardiovascular research. The technical difficulties of isolating individual cells from the zebrafish cardiovascular tissues have been limiting in studying their electrophysiological properties. Previous methods have been described for dissection of zebrafish hearts and isolation of ventricular cardiac myocytes. However, the isolation of zebrafish atrial and vascular myocytes for electrophysiological characterization was not detailed. This work describes new and modified enzymatic protocols that routinely provide isolated juvenile and adult zebrafish ventricular and atrial cardiomyocytes, as well as vascular smooth muscle (VSM) cells from the bulbous arteriosus, suitable for patch-clamp experiments. There has been no literary evidence of electrophysiological studies on zebrafish cardiovascular tissues isolated at embryonic and larval stages of development. Partial dissociation techniques that allow patch-clamp experiments on individual cells from larval and embryonic hearts are demonstrated.

The present protocol describes the acute isolation of viable cardiac and vascular smooth muscle cells from adult, juvenile, larval, and embryonic zebrafish (Danio rerio), suitable for electrophysiological studies.

Zebrafish have long been used as a model vertebrate organism in cardiovascular research. The technical difficulties of isolating individual cells from the ...

Adult Zebrafish Intestine Dissection: A Technique to Analyze Compound Absorption

Source: Jie J. et. al., <a href="https://www.jove.com/v/58366/" target="_blank">Oral Intubation of Adult Zebrafish: A Model for Evaluating Intestinal Uptake of Bioactive Compounds</a>. J. Vis. Exp. (2018).
This video describes the technique of isolating the intestine to evaluate the intestinal uptake of bioactive compounds. In this sample, we will isolate the intestine from an adult zebrafish to analyze the uptake of immunostimulants.

Source: Jie J. et. al., Oral Intubation of Adult Zebrafish: A Model for Evaluating Intestinal Uptake of Bioactive Compounds. J. Vis. Exp. (2018).
This ...

JoVE Encyclopedia of Experiments

Danio rerio (zebrafish)

Adult

Heart Dissection: A Method to Observe Zebrafish Cardiac Development

Source: Singleman C. et. al., <a href="https://www.jove.com/v/3165/" target="_blank">Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio</a>. J. Vis. Exp. (2011).
This video describes a technique of isolating the heart from a zebrafish at multiple developmental stages to study cardiac development, maturation, and ageing.

Source: Singleman C. et. al., Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio. J. Vis. Exp. (2011).
This video describes a technique ...

Dissection of Organs from the Adult Zebrafish

Summary

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Chapters in this video

Large-scale Zebrafish Embryonic Heart Dissection for Transcriptional Analysis

Dissection of Larval Zebrafish Gonadal Tissue

Dissection of the Adult Zebrafish Kidney

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

Analysis of Nephron Composition and Function in the Adult Zebrafish Kidney

Recovery of Adult Zebrafish Hearts for High-throughput Applications

Partial Hepatectomy in Adult Zebrafish

Isolation of Cardiac and Vascular Smooth Muscle Cells from Adult, Juvenile, Larval and Embryonic Zebrafish for Electrophysiological Studies

Adult Zebrafish Intestine Dissection: A Technique to Analyze Compound Absorption

Heart Dissection: A Method to Observe Zebrafish Cardiac Development