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

Este trabajo fue apoyado por el NIH subvención R01 HD050901 a MCM y estadounidenses beca postdoctoral Cancer Society # PF-05-041-01-DDC a TG.

<ol><li> Un pez cebra macho se diseca primero, seguido por un pez hembra. Antes de comenzar la disección, anestesiar a un pez en un 0,2% tricaína y la eutanasia que mediante la incubación en agua helada durante 15 minutos. </li><li> Comience suavemente acaricia el pescado seco en una toalla de papel y colocarlo sobre una alfombra de disección. Externamente, el pez cebra tiene una sola aleta dorsal, caudal y anal y emparejado aletas pectorales y pélvicas (Figura 1). <img src="/files/ftp_upload/1717/1717fig1.jpg" alt="Figura 1" /> Figura 1. Peces adultos machos con aletas de la etiqueta. </li><li> Pin el pescado a la lona a través de la disección de la parte carnosa de la cola y la parte ventral de la órbita del ojo. </li><li> Cortar la piel en el vientre del pez justo por delante de la aleta anal. Cortar la piel y el músculo subyacente a lo largo del vientre de la aleta anal a la opérculo (la cubierta dura más de las branquias) (Figura 2, paso 1). <img src="/files/ftp_upload/1717/1717fig2.jpg" alt="Figura 2" /> Figura 2. Pasos en la eliminación de la piel y el músculo de la pared del cuerpo para exponer los órganos internos. </li><li> A continuación, retire el opérculo y la aleta pectoral, incluyendo la faja pectoral (la región de espesor, del hueso en la base de la aleta) (Figura 2, paso 2). Cortar la piel y el músculo subyacente principio por encima de la ahora expuesta posteriormente enmalle a lo largo del lado de los peces y, después, a la aleta anal (Figura 2, paso 3). </li><li> Retire con cuidado la piel y el músculo subyacente desde el lado de los peces. Muchos de los órganos internos son visibles (Figura 3). <img src="/files/ftp_upload/1717/1717fig3.jpg" alt="Figura 3" /> Figura 3. Peces adultos masculinos con la pared del cuerpo y los músculos eliminado permitiendo la visualización de los órganos internos. </li><li> Los testículos son largos, los órganos blanco, a la par que se fija a la pared dorsal del cuerpo. Quitar un testículo y colócalo en un plato de PBS. Examinar los testículos con la luz reflejada para visualizar los túbulos seminíferos (Figura 4), que contienen quistes en las diferentes fases de las células germinales en desarrollo de spermatagonia de espermátidas (Leal et al., 2009). <img src="/files/ftp_upload/1717/1717fig4.jpg" alt="Figura 4" /> Figura 4. Disección de un testículo. Las estructuras redondeadas blancas son los túbulos seminíferos. </li><li> A continuación, retire el sistema gastrointestinal, desde la cavidad del cuerpo de los peces. El hígado puede ser identificado por su gran tamaño, la morfología lobulada, color tannish, y la vascularización extensa. La vesícula biliar, un saco verde transparente llena de líquido, y el bazo, de color rojo brillante, se encuentran dentro de las vísceras. </li><li> Separar el intestino del resto de los órganos y se extienden hacia fuera. Las regiones anterior, media y posterior del intestino se definen por la altura de los pliegues epiteliales (Wallace et al., 2005). </li><li> Coloque un pedazo de intestino en PBS y se observan los pliegues epiteliales con luz transmitida. </li><li> A continuación, examinar la vejiga natatoria. La vejiga natatoria se compone de una cámara posterior, que se conecta al esófago a través del conducto neumático y una cámara anterior, que se conecta al oído interno a través del aparato de Weber (Finney et al., 2006). </li><li> Retire y deseche la vejiga natatoria. Liberar a los peces y re-pin es la parte ventral hasta la disección del riñón, que se encuentra a lo largo de la pared dorsal del cuerpo. El riñón es una estructura transparente de color rosa asociado con la aorta dorsal y las células pigmentadas. El riñón se divide en regiones de la cabeza, cuerpo y cola (Figura 5). Diseccionar un pedazo de riñón y colocarlo en PBS. Separar el tejido del riñón para revelar los túbulos renales. <img src="/files/ftp_upload/1717/1717fig5.jpg" alt="Figura 5" /> Figura 5. Ubicación de la cabeza, cuerpo y cola del riñón a lo largo de la pared dorsal del cuerpo. </li><li> Diseccionar un pez hembra. Como se describió anteriormente, la eutanasia a los peces de agua fría y seque antes de sujetándola a la lona de disección. Quitar la piel de la cara de los peces como se demostró anteriormente. El ovario es una estructura bilobulada que se suspende en la cavidad del cuerpo por un mesovario vascularizado. </li><li> Extraer un lóbulo del ovario, lugar en PBS, y examinar con luz transmitida. Los ovocitos pueden ser objeto de burlas aparte utilizando agujas muy finas y por etapas (Figura 6, Selman et al., 1993). Ovocitos en estadio I es de aproximadamente 10 a 150 micras de tamaño y translúcido. Etapa II ovocitos son de 150 a 350 micras de tamaño y se define por la aparición de los gránulos corticales. Etapa III ovocitos son 350 a 750 micras de tamaño y son opacas, debido a la acumulación de yema de huevo. Ovocitos maduros etapa IV y V son translúcidos y, en general no se encuentran en los ovarios de las mujeres que recientemente han acoplado. <img src="/files/ftp_upload/1717/1717fig6.jpg" alt="Figura 6" /> Figura 6. Etapa en vivo I, II, III y los oocitos observados bajo unmicroscopio de disección con luz transmitida. </li><li> A continuación, analizar el corazón de los peces. El corazón se encuentra posterior y ventral de las branquias. Comience por cortar el corazón y todo el tejido circundante y colocarla en PBS. Diseccionar cuidadosamente el tejido que rodea el corazón, con cuidado de no dañar la delicada atrio. </li><li> Coloque el corazón disecado en solución de Ringer para observar el latido del corazón. Identificar el atrio, el ventrículo y bulbo arterioso (Figura 7, Hu et al., 2001). <img src="/files/ftp_upload/1717/1717fig7.jpg" alt="Figura 7" /> Figura 7. Disección del corazón adulto. </li><li> Completar la disección del cerebro mediante la eliminación de los peces. Comience por desclavar el pescado y la eliminación de la cabeza con una cuchilla de afeitar. Eliminar los tejidos blandos como sea posible de la parte ventral del cráneo con unas pinzas. Eliminar los ojos con unas tijeras pequeñas de primavera. Romper el cráneo y extraer el hueso de la parte ventral del cerebro. Ahora coloca la cabeza en un plato de PBS y quitar los huesos piel y el cráneo de la cara dorsal del cerebro. </li><li> Identificar los bulbos olfatorios, telencéfalo, habénula, techo óptico, cerebelo y la médula (Figura 8, Wullimann et al, 1996;. Schilling, 2002). <img src="/files/ftp_upload/1717/1717fig8.jpg" alt="Figura 8" /> Figura 8. Cerebro diseccionado adultos de pez cebra. Vista dorsal, anterior al inicio. </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>

Los experimentos con animales se realizaron de acuerdo con las directrices y normas establecidas por la Universidad de Pennsylvania Cuidado de Animales institucional y el empleo.

Reactivos: 0,2% tricaína (etil-3-aminobenzoato) 200 mg polvo tricaína 97,9 ml de agua DD ~ 1 ml 1 M Tris (pH 9) Ajustar el pH a 7.0 Tampón fosfato salino (PBS) 4,0 g de NaCl 0,1 g de KCl 100 ml 0,1 M tampón PO 4, pH 7,3 150 ml dH 2 O Solución Ringer 6,7 g de NaCl 0,2 g KCl 0,2 g de CaCl2 1,2 g de Hepes 1 LH 2 O Ajustar el pH a 7.2 Equipos: Plato de disección con estera Microscopía electrónica de Ciencias Catálogo # 70540 Vannas primavera tijera Herramientas de Bellas Ciencia Catálogo # 91500-09

dissection of organs from the adult zebrafish

En los últimos 20 años, el pez cebra se ha convertido en un organismo modelo de gran alcance para la comprensión del desarrollo de vertebrados y la enfermedad. Aunque el análisis experimental del embrión y la larva es muy amplia y la morfología ha sido bien documentada, las descripciones de la anatomía de adultos de pez cebra y los estudios de desarrollo de las estructuras y órganos adultos, junto con técnicas para trabajar con adultos que carecen. Los órganos de la larva de experimentar cambios significativos en su estructura general, la morfología y localización anatómica en la larva de la transición para adultos. Externamente, la larva transparente desarrolla su patrón característico de rayas pigmento y adultos emparejados aletas pélvicas, mientras que internamente, los órganos de someterse a un crecimiento masivo y remodelación. Además, la gónada bipotenciales primordio desarrolla en cualquiera de los testículos o los ovarios. Este protocolo se identifican muchos de los órganos del adulto y muestra los métodos para la disección del cerebro, las gónadas, el sistema gastrointestinal, el corazón y los riñones de los adultos de pez cebra. Los órganos diseccionados se pueden utilizar para la hibridación in situ, inmunohistoquímica, la histología, la extracción de ARN, el análisis de proteínas, y otras técnicas moleculares. Este protocolo ayudará en la ampliación de estudios en el pez cebra para incluir la remodelación de los órganos de las larvas, la morfogénesis de órganos específicos para las investigaciones de adultos y otros de los sistemas de órganos adultos.

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

Dissection of Organs from the Adult Zebrafish

Este protocolo describe un procedimiento para identificar y disecar los órganos de los adultos de pez cebra.

La disección de los órganos del pez cebra adultos

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

Research

JoVE Journal

Biology

104.5K vistas.  University of Pennsylvania-School of Medicine. Este protocolo describe un procedimiento para identificar y disecar los órganos de los adultos de pez cebra.

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.

A gran escala de pez cebra embrionario del corazón Disección de Análisis transcripcional

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 ...

Investigación

Biología del Desarrollo

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.

La disección de las larvas de pez cebra de tejidos gonadales

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.

La disección del riñón adultos de pez cebra

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

Biología

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.

La disección del corazón en las larvas, juveniles y adultos de pez cebra, 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.

Análisis de Composición Nephron y función en el riñón adultos de pez cebra

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.

Recuperación de adultos de pez cebra Corazones para aplicaciones de alto rendimiento

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.

Hepatectomía parcial en pez cebra adulto

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.

Aislamiento de células musculares lisas cardíacas y vasculares de peces cebra adultos, juveniles, larvales y embrionarios para estudios electrofisiológicos

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.

Disección del intestino del pez cebra adulto: una técnica para analizar la absorción de compuestos

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 (pez cebra)

Adulto

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.

Disección del corazón: un método para observar el desarrollo cardíaco del pez cebra

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