Name: dissection of organs from the adult zebrafish
Uploaded: 2010-03-04T16:45:00.000+00:00
Duration: 10 min 18 s
Description: Watch this Scientific Journal Video about Dissection of Organs from the Adult Zebrafish at JoVE.com

Ce travail a été soutenu par NIH R01 HD050901 à MCM et la bourse de recherche postdoctorale American Cancer Society # PF-05-041-01-DDC à TG.

<ol><li> Un poisson zèbre mâle sera disséqué en premier, suivi par un poisson femelle. Avant de commencer la dissection, anesthésier un poisson dans 0,2% tricaïne puis l&#39;euthanasier par incubation dans l&#39;eau glacée pendant 15 minutes. </li><li> Commencez par une légère tapotant le poisson sec sur une serviette en papier et en le plaçant sur un tapis de dissection. Extérieurement, le poisson zèbre ont dorsale, caudale et anale et jumelé nageoires pectorales et pelviennes (figure 1). <img src="/files/ftp_upload/1717/1717fig1.jpg" alt="Figure 1" /> Figure 1. Poissons mâles adultes avec des ailettes étiquetés. </li><li> Pin le poisson à travers le tapis disséquer la partie charnue de la queue et la partie ventrale de la cavité de l&#39;œil. </li><li> Snip la peau sur le ventre du poisson juste en avant de la nageoire anale. Coupez la peau et les muscles sous-jacents le long du ventre de la nageoire anale à l&#39;opercule (la couverture fort au cours des branchies) (figure 2, étape 1). <img src="/files/ftp_upload/1717/1717fig2.jpg" alt="Figure 2" /> Figure 2. Étapes de retirer la peau et les muscles de la paroi du corps pour exposer les organes internes. </li><li> Ensuite, retirez l&#39;opercule et la nageoire pectorale, y compris la ceinture scapulaire (l&#39;épaisseur, de la région osseuse à la base de la nageoire) (figure 2, étape 2). Coupez la peau et les muscles sous-jacents commencent au-dessus du présent exposé arrière des branchies le long du côté du poisson et ensuite jusqu&#39;à la nageoire anale (figure 2, étape 3). </li><li> Retirez soigneusement la peau et les muscles sous-jacents du côté du poisson. Beaucoup des organes internes sont maintenant visibles (figure 3). <img src="/files/ftp_upload/1717/1717fig3.jpg" alt="Figure 3" /> Figure 3. Adulte poissons mâles avec paroi musculaire du corps et enlevé permettant la visualisation des organes internes. </li><li> Les testicules sont longues, blanc, organes pairs qui sont attachés à la paroi dorsale du corps. Supprimer un testicule et le placer dans un plat de PBS. Examiner les testicules avec la lumière réfléchie de visualiser les tubules séminifères (Figure 4), qui contiennent des kystes aux divers stades de développement des cellules germinales à partir de spermatides spermatagonia (Leal et al., 2009). <img src="/files/ftp_upload/1717/1717fig4.jpg" alt="Figure 4" /> Figure 4. Dissection d&#39;un testicule. Les structures blanches arrondie sont les tubules séminifères. </li><li> Ensuite, retirez le système gastro-intestinal de la cavité abdominale du poisson. Le foie peut être identifié par sa grande taille, la morphologie lobé, de couleur tannish, et la vascularisation importante. La vésicule biliaire, d&#39;un vert translucide, sac rempli de liquide, et la rate, qui apparaît en rouge vif, se trouvent dans les viscères. </li><li> Séparée de l&#39;intestin du reste des organes et l&#39;étirer. L&#39;antérieure, les régions moyenne et postérieure de l&#39;intestin sont définis par la hauteur des plis épithéliaux (Wallace et al., 2005). </li><li> Placer un morceau de l&#39;intestin dans le PBS et d&#39;observer les replis épithéliaux en utilisant la lumière transmise. </li><li> Ensuite, examinez la vessie natatoire. La vessie natatoire se compose d&#39;une chambre postérieure, qui est relié à l&#39;œsophage par le conduit pneumatique, et une chambre antérieure, qui est relié à l&#39;oreille interne par l&#39;appareil de Weber (Finney et al., 2006). </li><li> Retirez et jetez la vessie natatoire. Détacher le poisson et re-broches c&#39;est la face ventrale à disséquer les reins, qui est située le long de la paroi dorsale du corps. Le rein est une structure associée rose translucide avec l&#39;aorte dorsale et les cellules pigmentées. Le rein est divisé en régions tête, le corps et la queue (figure 5). Disséquer un morceau du rein et de la placer dans du PBS. Démêler le tissu rénal de révéler les tubules rénaux. <img src="/files/ftp_upload/1717/1717fig5.jpg" alt="Figure 5" /> Figure 5. Localisation de la tête, le corps et la queue du rein le long de la paroi dorsale du corps. </li><li> Disséquer un poisson femelle. Comme décrit précédemment, euthanasier les poissons dans l&#39;eau glacée et sécher avant l&#39;épinglant sur le tapis de dissection. Enlevez la peau du côté des poissons comme précédemment démontré. L&#39;ovaire est une structure bilobée qui est suspendu dans la cavité du corps par un mésovarium vascularisé. </li><li> Retirer un lobe de l&#39;ovaire, le placer dans le PBS, et l&#39;examiner à la lumière transmise. Les ovocytes peuvent être taquiné dehors à l&#39;aide de fines aiguilles et ensuite mis en scène (figure 6, Selman et al., 1993). Phase I ovocytes sont environ 10 - 150 microns dans la taille et translucides. Etape II ovocytes sont environ 150 - 350 microns dans la taille et définie par l&#39;apparition de granules corticaux. Etape III ovocytes sont de 350 à 750 microns et sont opaques à cause de l&#39;accumulation de jaune d&#39;oeuf. Mature étape ovocytes IV et V sont translucides et généralement ne se trouve pas dans les ovaires des femelles qui ont récemment estimé. <img src="/files/ftp_upload/1717/1717fig6.jpg" alt="Figure 6" /> Figure 6. Étape en direct I, II, III et des ovocytes observé sous undisséquant microscope à lumière transmise. </li><li> Ensuite, disséquer le cœur du poisson. Le cœur est situé postérieure et ventrale de la branchie. Commencez par découper le cœur et l&#39;ensemble des tissus environnants et de la placer dans du PBS. Soigneusement disséquer loin le tissu entourant le coeur, en faisant attention à ne pas endommager l&#39;atrium délicate. </li><li> Placez le coeur disséqué dans la solution de Ringer à observer les battements du cœur. Identifier l&#39;atrium, ventricule et bulbe artériel (figure 7, Hu et al., 2001). <img src="/files/ftp_upload/1717/1717fig7.jpg" alt="Figure 7" /> Figure 7. Disséqué coeur adulte. </li><li> Remplissez la dissection en enlevant le cerveau des poissons. Commencez par désépinglage le poisson et enlever la tête avec une lame de rasoir. Retirez le tissu mou autant que possible de la face ventrale du crâne avec une pince. Retirez les yeux à l&#39;aide des ciseaux petit ressort. Pause ouvrir le crâne et enlever l&#39;os de la face ventrale du cerveau. Maintenant, placez la tête dans un plat de PBS et enlever les os du crâne à partir de la peau et la face dorsale du cerveau. </li><li> Identifier les bulbes olfactifs, télencéphale, habénula, tectum optique, le cervelet et le bulbe (figure 8, Wullimann et al, 1996;. Schilling, 2002). <img src="/files/ftp_upload/1717/1717fig8.jpg" alt="Figure 8" /> Figure 8. Disséqué le cerveau de poisson zèbre adulte. Vue dorsale, antérieure vers le haut. </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>

Expériences sur les animaux ont été effectuées en conformité avec les directives et règlements établis par l&#39;Université de Pennsylvanie des soins des animaux et du Comité institutionnel d&#39;utilisation.

Réactifs: 0,2% (éthyl 3-aminobenzoate) tricaïne 200 mg de poudre tricaïne 97,9 ml d&#39;eau JJ ~ 1 ml 1 M Tris (pH 9) Ajuster le pH à 7,0 Tampon phosphate salin (PBS) 4,0 g de NaCl 0,1 g de KCl 100 ml à 0,1 M de tampon PO 4, pH 7,3 150 ml dH 2 O Solution de Ringer 6.7g de NaCl 0.2g KCl 0.2g CaCl 2 Hepes 1,2 g 1 LH 2 O Ajuster le pH à 7,2 Equipement: Dissection plat avec tapis Sciences Electron Microscopy Le catalogue # 70540 Vannas des ciseaux à ressort Outils Fine Science Le catalogue # 91500-09

dissection of organs from the adult zebrafish

Au cours des 20 dernières années, le poisson-zèbre est devenu un organisme modèle puissant pour comprendre le développement des vertébrés et de la maladie. Bien que l&#39;analyse expérimentale de l&#39;embryon et la larve est vaste et la morphologie a été bien documenté, les descriptions de l&#39;anatomie de poisson zèbre pour adultes et des études sur le développement des structures pour adultes et les organes, ainsi que des techniques pour travailler avec les adultes sont absents. Les organes de la larve subissent d&#39;importants changements dans leur structure générale, la morphologie et la localisation anatomique lors de la larve de la transition pour adultes. Extérieurement, la larve transparente développe son modèle caractéristique de pigments pour adultes rayé et jumelé nageoires pelviennes, tandis qu&#39;à l&#39;intérieur, les organes subissent une croissance massive et le remodelage. En outre, le primordium bipotentielles gonades se développe en testicule ou l&#39;ovaire soit. Ce protocole identifie la plupart des organes de l&#39;adulte et démontre des méthodes pour la dissection du cerveau, les gonades, le système gastro-intestinal, le cœur et les reins du poisson-zèbre adulte. Les organes disséqués peut être utilisé pour l&#39;hybridation in situ, immunohistochimie, l&#39;histologie, l&#39;extraction d&#39;ARN, l&#39;analyse des protéines, et d&#39;autres techniques moléculaires. Ce protocole aidera à l&#39;élargissement des études dans le poisson zèbre pour inclure le remodelage des organes larvaires, la morphogenèse des organes spécifiques aux adultes et d&#39;autres enquêtes sur les systèmes d&#39;organes adultes.

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

Dissection of Organs from the Adult Zebrafish

Ce protocole décrit une procédure pour identifier et disséquer les organes de l&#39;zèbre adulte.

Dissection d&#39;organes sur le poisson zèbre pour adultes

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

dissection-of-organs-from-the-adult-zebrafish

Universidad San Sebastian

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JoVE Journal

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104.5K vues.  University of Pennsylvania-School of Medicine. Ce protocole décrit une procédure pour identifier et disséquer les organes de l'zèbre adulte.

Vidéo: 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.

À grande échelle de poisson zèbre embryonnaire Coeur Dissection d&#39;analyse transcriptionnelle

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 ...

Recherche

Biologie du développement

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.

Dissection de tissu gonadique Larval Zebrafish

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.

Dissection du rein Zebrafish adultes

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

Biologie

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.

Dissection du cœur chez les larves, le poisson zèbre juvéniles et adultes, Danio rerio</em

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.

Analyse de néphron Composition et fonction du rein poisson zèbre adulte

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.

Recouvrement des adultes coeurs de poisson zèbre à haut débit pour des applications

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.

Hépatectomie partielle chez le poisson zèbre adulte

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.

Isolement de cellules musculaires lisses cardiaques et vasculaires provenant de poissons-zèbres adultes, juvéniles, larvaires et embryonnaires pour des études électrophysiologiques

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.bdigitaluss.remotexs.co/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.

Dissection intestinale de poisson-zèbre adulte : une technique pour analyser l’absorption des composés

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.bdigitaluss.remotexs.co/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.

Dissection cardiaque : une méthode pour observer le développement cardiaque du poisson-zèbre

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 d'organes sur le poisson zèbre pour adultes

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À grande échelle de poisson zèbre embryonnaire Coeur Dissection d'analyse transcriptionnelle