Nos gustaría agradecer a nuestro asesor, el Dr. Eva Marder. Esta investigación fue financiada por el Instituto Nacional de Salud subvenciones NS17813 a Eva Marder y NS059255 a AE T.

<ol><li> Antes de la disección de la langosta debe ser enterrado en el hielo durante 30 minutos para anestesiar a los animales. </li><li> El sexo de la langosta puede ser determinado observando la forma de la primera swimmeret. Los varones tienen swimmerets óseas y las hembras tienen swimmerets plumas. <img src="/files/ftp_upload/1320/1320fig1.png" alt="" /></li><li> Aislar el cefalotórax. <ol><li> Use un movimiento de torsión para estafar a las garras delanteras. </li><li> Retire el abdomen (parte de la cola) mediante el uso de las tijeras grandes para hacer un corte profundo entre el cefalotórax y abdomen. Esto reduce los tractos de fibras gigantes que controlan la respuesta voltear la cola. A continuación, proceder a cortar el abdomen. </li><li> Quite las antenas por el recorte de ellos con unas tijeras, y giro de las piernas y los maxilípedos terceros. </li></ol></li><li> Retire el caparazón de todo el estómago, (es decir, la región de la cabeza entre la ranura del cuello uterino y la tribuna). <ol><li> A partir del borde lateral de la sección de pinzas para el uso y la espátula para retirar con cuidado el caparazón duro, dejando la hipodermis subyacente rojizo intacto. La espátula es útil para la separación de las capas para que el caparazón se rompió con la gubia. </li><li> Avanzar hacia los ojos a encontrar bilateral ligeramente calcificada placas pequeño óvalo (IIIa huesecillos). Lo más cerca posible a estos huesecillos, separar con cuidado el músculo de los huesecillos del oído con las tijeras pequeñas. </li><li> Usando una espátula y pinzas para quitar caparazón de entre los ojos y la porción caudal (aproximadamente 1 centímetro) de la tribuna. </li></ol></li><li> Separe la cara (región, incluyendo los ojos y rostro). <ol><li> Con la tijera pequeña hacer un corte horizontal a través del tejido más rostral expuestos en la tribuna para separarlo del exoesqueleto. A continuación, utilice el borde de la tijera y / o una espátula para rodar con cuidado el tejido caudal, justo pasado el nivel de la cadena de huesecillos IIIa. </li><li> A partir dorsal, use tijeras pequeñas para separar el tejido restante del exoesqueleto cara. Corte lo más cerca del caparazón duro como sea posible, se detiene cuando usted consigue cerca del centro y el apego de la boca. </li><li> En la vista ventral de las mandíbulas deben ser fácilmente visibles. Separe maxilípedos restantes. </li><li> Localizar los labios suaves en las mandíbulas. </li><li> Haga cortes laterales para separar la cara mientras se mantiene el labio une al estómago. (Hay una parte ligeramente curvada azul boca externa que pueda ser utilizado como una guía.) </li></ol></li><li> Desmontar y retirar el estómago. <ol><li> Mantenga la langosta en la palma de su mano, la cara ventral y para que el extremo anterior se dirige hacia el codo. </li><li> Coloque una pinza hemostática en el labio y dejar que la gravedad tire de la hemostasia y los labios hacia abajo y lejos de la pared ventral del cuerpo. </li><li> Con la tijera pequeña cortada mandíbulas. </li><li> Localice la cara dorsal de la pared ventral del cuerpo, que forma un duro &quot;plataforma&quot; para el labio ventral y el estómago. Corte justo debajo de la &quot;plataforma&quot;, muy cerca de la pared del cuerpo, a través de todo medio-lateral la línea media. Toda la parte ventral del estómago ahora debe ser separada de la pared del cuerpo. </li><li> A su vez de la langosta de manera que la parte dorsal está arriba. Corte a lo largo del borde rostral del surco cervical, donde se extirpó el caparazón, y tira suavemente el estómago hacia fuera. </li><li> Separe los intestinos extra que permanecer unidos hasta el estómago. </li></ol></li><li> Abra el estómago, y prepararlo para la fijación. <ol><li> Mantener el estómago en la palma de la mano con la hipodermis hacia abajo. La pinza hemostática aún debe atribuirse a los labios y debe estar entre los dedos. </li><li> Utilice unas tijeras pequeñas para hacer un poco anterior-posterior corte de la apertura de la boca (por debajo del labio) a través de la parte ventral del píloro. </li><li> Para abrir aún más el estómago, hacer dos cortes laterales a lo largo de la cadena de huesecillos que atraviesa el centro del estómago. </li><li> Solución salina fría puede ser vertido en el estómago para mejorar la visibilidad y expulsar el contenido del estómago. </li><li> A fin de permitir la preparación para poner completamente en el plato de la disección de utilizar las tijeras pequeñas para recortar los tres dientes gástrico. </li></ol></li><li> Pin del estómago hacia abajo en el plato de la disección. <ol><li> Coloque el estómago, hipodermis hacia arriba, en un plato de la disección de solución salina. </li><li> Coloque pasadores de insectos en los labios y las cuatro esquinas creado cuando el estómago se abrió. Por las esquinas anterior pines altura y cerca de la orilla, no molestar a los ganglios de la comisura inferior o nervios del esófago. Para los pines esquinas posteriores lugar un poco alta para no molestar el músculo dilatador del píloro. </li></ol></li></ol>

<ol>
	<li>Marder, E., Bucher, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+pattern+generators+and+the+control+of+rhythmic+movements.">Central pattern generators and the control of rhythmic movements.</a> Curr. Biol. 11, R986-R996 (2001).</li><li>Nusbaum, M. P., Beenhakker, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+small-systems+approach+to+motor+pattern+generation.">A small-systems approach to motor pattern generation.</a> Nature. 417, 343-350 (2002).</li><li>Clemens, S., Massabuau, J. C., Legeay, A., Meyrand, P., Simmers, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+modulation+of+interacting+central+pattern+generators+in+lobster+stomatogastric+ganglion:+influence+of+feeding+and+partial+pressure+of+oxygen.">In vivo modulation of interacting central pattern generators in lobster stomatogastric ganglion: influence of feeding and partial pressure of oxygen.</a> J. Neurosci. 18, 2788-2799 (1998).</li><li>Tobin, A. E., Bierman, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homarus+americanus+stomatogastric+nervous+system+dissection.">Homarus americanus stomatogastric nervous system dissection.</a> J Vis Exp. 27, (2009).</li><li>Maynard, D. M., Dando, M. 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+structure+of+the+stomatogastric+neuromuscular+system+in+Callinectes+sapidus,+Homarus+Americanus+and+Panulirus+argus+(Decapoda+crustacean).">The structure of the stomatogastric neuromuscular system in Callinectes sapidus, Homarus Americanus and Panulirus argus (Decapoda crustacean).</a> Philos. Trans. R. Soc. Lond. B Biol. Sci. 268, 161-220 (1974).</li><li>Herrick, F. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Natural+history+of+the+American+Lobster.">Natural history of the American Lobster.</a> U.S. Bur. Fish. , 219-252 (1909).</li><li>Marder, E., Bucher, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+circuit+dynamics+using+the+stomatogastric+nervous+system+of+lobsters+and+crabs.">Understanding circuit dynamics using the stomatogastric nervous system of lobsters and crabs.</a> Annu. Rev. Physiol. 69, 291-316 (2007).</li></ol>

En nuestro laboratorio, la disección del estómago de langosta se hace para tener acceso al sistema nervioso stomatogastric, una red que controla las contracciones rítmicas del estómago. La anatomía del estómago la langosta ha sido bien descrita previamente por 5-7, y se recomienda familiarizarse con su anatomía. Si lo hace, reducirá el riesgo de dañar los delicados nervios y músculos. Una vez completada la disección del estómago, varias porciones de la STNS puede ser m...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Homarus americanus</td>
<td>&nbsp;</td>
<td>Commercial Lobster</td>
<td>&nbsp;</td>
<td>Can be purchased from any lobster supplier, We specifically request &#x201C;chicken&#x201D; lobsters.</td>
</tr>
<tr>
<td>Black Sylgard 170</td>
<td>&nbsp;</td>
<td>Dow Corning</td>
<td>170</td>
<td>&nbsp;</td>
<tr>
<td>Preparation Dish</td>
<td>&nbsp;</td>
<td>Fischer </td>
<td>08-807</td>
<td>Fill 2" with Black Sylgard 170.</td>
</tr>
<tr>
<td>Fine Scissors</td>
<td>&nbsp;</td>
<td>FST</td>
<td>14090-09</td>
<td>&nbsp;</td>
<tr>
<td>Insect Pins size 6</td>
<td>&nbsp;</td>
<td>FST</td>
<td>26000-65</td>
<td>&nbsp;</td>
<tr>
<td>Mayo Scissors</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>This is a common product offered by many suppliers of dissecting tools.</td>
</tr>
<tr>
<td>Micro-Spatula</td>
<td>&nbsp;</td>
<td>Ward Sci.</td>
<td>15 V 4313</td>
<td>This is a very common product offered by many suppliers of lab tools.</td>
</tr>
<tr>
<td>Rongeurs</td>
<td>&nbsp;</td>
<td>FST</td>
<td>&nbsp;</td>
<td>16000-14</td>
</tr>
<tr>
<td>Plastic hemostat</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>This is a common product offered by many suppliers of dissecting or jewelry making tools.</td>
</tr>		</tbody>
		</table>
		</div>

gross dissection of the stomach of the lobster, homarus americanus

El estómago de la langosta americana (<i&gt; Homarus americanus</i&gt;) Se encuentra en el cefalotórax, entre la tribuna y el surco del cuello uterino. El extremo anterior del estómago se define por la apertura de la boca y el extremo posterior de la parte inferior del píloro. A lo largo de la parte dorsal del estómago se encuentra el sistema nervioso stomatogastric (STNS). Este sistema nervioso, que contiene las redes rítmica que subyacen al comportamiento de alimentación, es un sistema modelo establecido para el estudio de las redes de la generación del ritmo y la neuromodulación<sup&gt; 1,2</sup&gt;. Si bien es posible estudiar este sistema<i&gt; In vivo</i<sup&gt; 3</sup&gt;, El STNS sigue produciendo su actividad rítmica cuando se aíslan<i&gt; In vitro</i&gt;. Para el estudio de este sistema<i&gt; In vitro</i&gt; El estómago debe ser quitada del animal. En este artículo se describe cómo el vídeo del estómago puede ser diseccionada de la langosta americana. En un artículo de video que lo acompaña<sup&gt; 4</sup&gt; Que demuestran cómo el STNS puede ser aislado del estómago.

Watch this Scientific Journal Video about Gross Dissection of the Stomach of the Lobster, Homarus Americanus at JoVE.com

Gross Dissection of the Stomach of the Lobster, Homarus Americanus

Se describe la disección bruto del estómago de la langosta americana (<i&gt; Homarus americanus</i&gt;).

La disección bruto del estómago de la langosta, Homarus americanus</em

The stomach of the American lobster (Homarus americanus) is located in the cephalothorax, between the rostrum and the cervical groove. The anterior end of ...

gross-dissection-of-the-stomach-of-the-lobster-homarus-americanus

Research

JoVE Journal

Biology

Gross Dissection of the Stomach of the Lobster, Homarus Americanus

14.4K vistas.  Brandeis. Se describe la disección bruto del estómago de la langosta americana ( Homarus americanus).

Video: Gross Dissection of the Stomach of the Lobster, Homarus Americanus

Micro-dissection of Rat Brain for RNA or Protein Extraction from Specific Brain Region

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates micro-dissection of four major brain regions include olfactory bulb, frontal cortex, striatum and hippocampus in fresh rat brain tissue. Useful tips for quick removal of respective regions to avoid RNA and protein degradation of the tissue are given.

Micro-disección de cerebro de rata para el ARN o la extracción de proteínas a partir de región específica del cerebro

Micro-dissection of rat brain into various regions is extremely important for the study of different neurodegenerative diseases. This video demonstrates ...

Investigación

Biología

Homarus Americanus Stomatogastric Nervous System Dissection

With the goal of understanding how nervous systems produce activity and respond to the environment, neuroscientists turn to model systems that exhibit the activity of interest and are accessible and amenable to experimental methods. The stomatogastric nervous system (STNS) of the American lobster (Homarus americanus; also know was the Atlantic or Maine lobster) has been established as a model system for studying rhythm generating networks and neuromodulation of networks. The STNS consists of 3 anterior ganglia (2 commissural ganglia and an oesophageal ganglion), containing modulatory neurons that project centrally to the stomatogastric ganglion (STG). The STG contains approximately 30 neurons that comprise two central pattern generating networks, the pyloric and gastric networks that underlie feeding behaviors in crustaceans1,2. While it is possible to study this system in vivo3, the STNS continues to produce its rhythmic activity when isolated in vitro. Physical isolation of the STNS in a dish allows for easy access to the somata in the ganglia for intracellular electrophysiological recordings and to the nerves of the STNS for extracellular recordings. Isolating the STNS is a two-part process. The first part, dissecting the stomach from the animal, is described in an accompanying video article4. In this video article, fine dissection techniques are used to isolate the STNS from the stomach. This procedure results in a nervous system preparation that is available for electrophysiological recordings.

Homarus Americanus Stomatogastric Nervous System Dissection

We describe the fine dissection of the stomatogastric nervous system from the stomach of the American lobster (Homarus americanus).

Homarus americanus Stomatogastric La disección del sistema nervioso

With the goal of understanding how nervous systems produce activity and respond to the environment, neuroscientists turn to model systems that exhibit the ...

Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish (Danio rerio)

Neurosensory epithelia in the inner ear are the crucial structures for hearing and balance functions. Therefore, it is important to understand the cellular and molecular features of the epithelia, which are mainly composed of two types of cells: hair cells (HCs) and supporting cells (SCs). Here we choose to study the inner ear sensory epithelia in adult zebrafish not only because the epithelial structures are highly conserved in all vertebrates studied, but also because the adult zebrafish is able to regenerate HCs, an ability that mammals lose shortly after birth. We use the inner ear of adult zebrafish as a model system to study the mechanisms of inner ear HC regeneration in adult vertebrates that could be helpful for clinical therapy of hearing/balance deficits in human as a result of HC loss.
Here we demonstrate how to do gross and fine dissections of inner ear sensory epithelia in adult zebrafish. The gross dissection removes the tissues surrounding the inner ear and is helpful for preparing tissue sections, which allows us to examine the detailed structure of the sensory epithelia. The fine dissection cleans up the non-sensory-epithelial tissues of each individual epithelium and enables us to examine the heterogeneity of the whole epithelium easily in whole-mount epithelial samples.

Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish Danio rerio

The inner ear sensory epithelium of adult zebrafish is a good model system for understanding the mechanisms of hair cell regeneration in adult vertebrates. This protocol demonstrates the fine dissection of the epithelia, through which we can get tissue samples for studying the regenerative events at cellular and subcellular levels.

La disección fina y gruesa de los epitelios sensoriales del oído interno en adultos de pez cebra ( Danio rerio)

Neurosensory epithelia in the inner ear are the crucial structures for hearing and balance functions. Therefore, it is important to understand the ...

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.

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

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 ...

Using SCOPE to Identify Potential Regulatory Motifs in Coregulated Genes

SCOPE is an ensemble motif finder that uses three component algorithms in parallel to identify potential regulatory motifs by over-representation and motif position preference1. Each component algorithm is optimized to find a different kind of motif. By taking the best of these three approaches, SCOPE performs better than any single algorithm, even in the presence of noisy data1. In this article, we utilize a web version of SCOPE2 to examine genes that are involved in telomere maintenance. SCOPE has been incorporated into at least two other motif finding programs3,4 and has been used in other studies5-8. 
The three algorithms that comprise SCOPE are BEAM9, which finds non-degenerate motifs (ACCGGT), PRISM10, which finds degenerate motifs (ASCGWT), and SPACER11, which finds longer bipartite motifs (ACCnnnnnnnnGGT). These three algorithms have been optimized to find their corresponding type of motif. Together, they allow SCOPE to perform extremely well.
Once a gene set has been analyzed and candidate motifs identified, SCOPE can look for other genes that contain the motif which, when added to the original set, will improve the motif score. This can occur through over-representation or motif position preference. Working with partial gene sets that have biologically verified transcription factor binding sites, SCOPE was able to identify most of the rest of the genes also regulated by the given transcription factor.
Output from SCOPE shows candidate motifs, their significance, and other information both as a table and as a graphical motif map. FAQs and video tutorials are available at the SCOPE web site which also includes a "Sample Search" button that allows the user to perform a trial run. 
Scope has a very friendly user interface that enables novice users to access the algorithm's full power without having to become an expert in the bioinformatics of motif finding. As input, SCOPE can take a list of genes, or FASTA sequences. These can be entered in browser text fields, or read from a file. The output from SCOPE contains a list of all identified motifs with their scores, number of occurrences, fraction of genes containing the motif, and the algorithm used to identify the motif. For each motif, result details include a consensus representation of the motif, a sequence logo, a position weight matrix, and a list of instances for every motif occurrence (with exact positions and "strand" indicated). Results are returned in a browser window and also optionally by email. Previous papers describe the SCOPE algorithms in detail1,2,9-11.

A straight-forward and robust method to identify potential regulatory motifs in co-regulated genes is presented. SCOPE does not require any user parameters and returns motifs that represent excellent candidates for regulatory signals. The identification of such regulatory signals helps to understand the underlying biology.

Usando alcance para identificar posibles motivos de reglamentación en los genes coregulated

SCOPE is an ensemble motif finder that uses three component algorithms in parallel to identify potential regulatory motifs by over-representation and ...

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 ...

Dissection and Flat-mounting of the Threespine Stickleback Branchial Skeleton

The posterior pharyngeal segments of the vertebrate head give rise to the branchial skeleton, the primary site of food processing in fish. The morphology of the fish branchial skeleton is matched to a species' diet. Threespine stickleback fish (Gasterosteus aculeatus) have emerged as a model system to study the genetic and developmental basis of evolved differences in a variety of traits. Marine populations of sticklebacks have repeatedly colonized countless new freshwater lakes and creeks. Adaptation to the new diet in these freshwater environments likely underlies a series of craniofacial changes that have evolved repeatedly in independently derived freshwater populations. These include three major patterning changes to the branchial skeleton: reductions in the number and length of gill raker bones, increases in pharyngeal tooth number, and increased branchial bone lengths. Here we describe a detailed protocol to dissect and flat-mount the internal branchial skeleton in threespine stickleback fish. Dissection of the entire three-dimensional branchial skeleton and mounting it flat into a largely two-dimensional prep allows for the easy visualization and quantification of branchial skeleton morphology. This dissection method is inexpensive, fast, relatively easy, and applicable to a wide variety of fish species. In sticklebacks, this efficient method allows the quantification of skeletal morphology in genetic crosses to map genomic regions controlling craniofacial patterning.

The branchial skeleton, including gill rakers, pharyngeal teeth, and branchial bones, serves as the primary site of food processing in most fish. Here we describe a protocol to dissect and flat-mount this internal skeleton in threespine sticklebacks. This method is also applicable to a variety of other fish species.

La disección y-montaje plana del esqueleto branquial de tres espinas Stickleback

The posterior pharyngeal segments of the vertebrate head give rise to the branchial skeleton, the primary site of food processing in fish. The morphology ...

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

Lipids are essential for animal development and physiological homeostasis. Dysregulation of lipid metabolism results in various developmental defects and diseases, such as obesity and fatty liver. Usually, lipids are stored in lipid droplets, which are the multifunctional lipid storage organelles in cells. Lipid droplets vary in size and number in different tissues and under different conditions. It has been reported that lipid droplets are tightly controlled through regulation of its biogenesis and degradation. In Drosophila melanogaster, the oenocyte is an important tissue for lipid metabolism and has been recently identified as a human liver analogue regarding lipid mobilization in response to stress. However, the mechanisms underlying the regulation of lipid droplet metabolism in oenocytes remain elusive. To solve this problem, it is of utmost importance to develop a reliable and sensitive method to directly visualize lipid droplet dynamic changes in oenocytes during development and under stressful conditions. Taking advantage of the lipophilic BODIPY 493/503, a lipid droplet-specific fluorescent dye, described here is a detailed protocol for the dissection and subsequent lipid droplet staining in the oenocytes of Drosophila larvae in response to starvation. This allows for qualitative analysis of lipid droplet dynamics under various conditions by confocal microscopy. Furthermore, this rapid and highly reproducible method can also be used in genetic screens for indentifying novel genetic factors involving lipid droplet metabolism in oenocytes and other tissues.

Dissection and Lipid Droplet Staining of Oenocytes in Drosophila Larvae

Presented here are detailed methods for the dissection and lipid droplet staining of oenocytes in Drosophila larvae using BODIPY 493/503, a lipid droplet-specific fluorescent dye.

Disección y tinción de gotas de lípidos de los ovenocitos en larvas de Drosophila

Lipids are essential for animal development and physiological homeostasis. Dysregulation of lipid metabolism results in various developmental defects and ...

Dissection of the Endolymphatic Sac from Mice

The study of mutant mouse models of human hearing and balance disorders has unraveled many structural and functional changes which may contribute to the human phenotypes. Although important progress has been done in the understanding of the development and function of the neurosensory epithelia of the cochlea and vestibula, limited knowledge is available regarding the development, cellular composition, molecular pathways and functional characteristics of the endolymphatic sac. This is, in large part, due to the difficulty of visualizing and microdissecting this tissue, which is an epithelium comprised of only one cell layer. The study presented here describes an approach to access and microdissect the endolymphatic sac from the wild-type mouse inner ear at different ages. The result of a similar dissection is shown in a pendrin-deficient mouse model of enlargement of the vestibular aqueduct. A transgenic mouse with a fluorescent endolymphatic sac is presented. This reporter mouse can be used to readily visualize the endolymphatic sac with limited dissection and determine its size. It can also be used as an educational tool to teach how to dissect the endolymphatic sac. These dissection procedures should facilitate further characterization of this understudied part of the inner ear.

<meta charset="utf8"></head><body>Disección del saco endolinfático de ratones

This study describes how to dissect the endolymphatic sac from the inner ear of the mouse at different ages. The result of a similar dissection is shown in an Slc26a4-null mouse model of enlargement of the endolymphatic sac. A transgenic mouse with a fluorescent reporter expressed in the endolymphatic sac is presented as a model to readily visualize the endolymphatic sac, and educational tool.

Disección del saco endolinfático de ratones

The study of mutant mouse models of human hearing and balance disorders has unraveled many structural and functional changes which may contribute to the ...