Financiación proporcionada por las Becas de Ribble G. en el Departamento de Biología, los investigadores jóvenes y Kentucky Pregrado Educación Eureka! Oficina (Univ. de KY).

<ol>
	<li>Ashton, K., Wagoner, A. P., Carrillo, R., Gibson, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+trait+loci+for+the+monoamine-related+traits+heart+rate+and+headless+behavior+in+Drosophila+melanogaster.">Quantitative trait loci for the monoamine-related traits heart rate and headless behavior in Drosophila melanogaster.</a> Genetics. 157, 283-294 (2001).</li><li>Azpiazu, N., Frasch, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tinman+and+Bagpipe:+Two+homeo+box+genes+that+determine+cell+fates+in+the+dorsal+mesoderm+of+Drosophila.">Tinman and Bagpipe: Two homeo box genes that determine cell fates in the dorsal mesoderm of Drosophila.</a> Genes &#38; Development. 7, 1325-1340 (1993).</li><li>Badre, N. H., Martin, M. E., Cooper, R. 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Biol. 202, 3037-3048 (1999).</li><li>Bier, E., Bodmer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila,+an+emerging+model+for+cardiac+disease.">Drosophila, an emerging model for cardiac disease.</a> Gene. 342, 1-11 (2004).</li><li>Bodmer, 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+gene+tinman+is+required+for+specification+of+the+heart+and+visceral+muscles+in+Drosophila.">The gene tinman is required for specification of the heart and visceral muscles in Drosophila.</a> Development. 118, 719-729 (1993).</li><li>Bodmer, R., Jan, L. Y., Jan, Y. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+homeobox-containing+gene,+msh-2,+is+transiently+expressed+early+during+mesoderm+formation+of+Drosophila.">A new homeobox-containing gene, msh-2, is transiently expressed early during mesoderm formation of Drosophila.</a> Development. 110, 661-669 (1990).</li><li>Cooper, A. S., Cooper, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monitoring+activity+of+Drosophila+larvae:+Impedance+&#38;+video+microscopy+measures.">Monitoring activity of Drosophila larvae: Impedance &#38; video microscopy measures.</a> Drosophila Information Service. 87, 85-87 (2004).</li><li>Dasari, S., Cooper, R. 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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modified+minimal+hemolymph-like+solution,+HL3.1,+for+physiological+recordings+at+the+neuromuscular+junctions+of+normal+and+mutant+Drosophila+larvae.">A modified minimal hemolymph-like solution, HL3.1, for physiological recordings at the neuromuscular junctions of normal and mutant Drosophila larvae.</a> J. Neurogenetics. 18, 377-402 (2004).</li><li>Ganetzky, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+analysis+of+ion+channel+dysfunction+in+Drosophila.">Genetic analysis of ion channel dysfunction in Drosophila.</a> Kidney International. 3, 766-771 (2000).</li><li>GG, G. u. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+analysis+of+heartbeat+in.">Pharmacological analysis of heartbeat in.</a> Drosophila. J. Neurobiol. 28, 269-280 (1995).</li><li>He, B., Adler, P. 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B. 167, 89-97 (1997).</li><li>Johnson, E., Ringo, J., Bray, N., Dowse, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+and+pharmacological+identification+of+ion+channels+central+to+the+Drosophila+cardiac+pacemaker.">Genetic and pharmacological identification of ion channels central to the Drosophila cardiac pacemaker.</a> J. Neurogenetics. 12, 1-24 (1998).</li><li>Johnson, E., Ringo, J., Dowse, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Native+and+heterologous+neuropeptides+are+cardioactive+in+Drosophila+melanogaster.">Native and heterologous neuropeptides are cardioactive in Drosophila melanogaster.</a> J. Insect Physiol. 1, 1229-1236 (2000).</li><li>Johnson, E., Sherry, T., Ringo, J., Dowse, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+the+cardiac+pacemaker+of+Drosophila:+cellular+mechanisms.">Modulation of the cardiac pacemaker of Drosophila: cellular mechanisms.</a> J. Comp. Physiol. B, Biochem., Systemic, Environmental Physiol. 172, 227-236 (2002).</li><li>Lewis, 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=A+new+standard+food+medium.">A new standard food medium.</a> Drosophila Inform. Serv. 34, 117-117 (1960).</li><li>Miller, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+circulation+in+insects.">Control of circulation in insects.</a> General Pharmacol. 29, 23-38 (1997).</li><li>Molina, M. R., Cripps, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ostia,+the+inflow+tracts+of+the+Drosophila+heart,+develop+from+a+genetically+distinct+subset+of+cardial+cells.">Ostia, the inflow tracts of the Drosophila heart, develop from a genetically distinct subset of cardial cells.</a> Mechanisms of Development. 118, 51-59 (2001).</li><li>Muller, K. J., Nicholls, J. G., Stent, G. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Neurobiology of the Leech. , 254-254 (1981).</li><li>Nichols, R., Kaminski, S., Walling, E., Zornik, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regulating+the+activity+of+a+cardioacceleratory+peptide.">Regulating the activity of a cardioacceleratory peptide.</a> Peptides. 20, 1153-1158 (1999).</li><li>Ocorr, K., Akasaka, T., Bodmer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-related+cardiac+disease+model+of+Drosophila.">Age-related cardiac disease model of Drosophila.</a> Mechanisms of Ageing and Development. 128, 112-116 (2007a).</li><li>Ocorr, K. A., Crawley, T., Gibson, G., Bodmer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+variation+for+cardiac+dysfunction+in+Drosophila.">Genetic variation for cardiac dysfunction in Drosophila.</a> PLoS ONE. 2, e601-e601 (2007).</li><li>Papaefthmiou, C., Theophilidis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+in+vitro+method+for+recording+the+electrical+activity+of+the+isolated+heart+of+the+adult+Drosophila+melanogaster.">An in vitro method for recording the electrical activity of the isolated heart of the adult Drosophila melanogaster.</a> In Vitro Cellular &#38; Developmental Biol. Animal. 37, 445-449 (2001).</li><li>Peron, S., Zordan, M. A., Magnabosco, A., Reggiani, C., Megighian, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+action+potential+to+contraction:+Neural+control+and+excitation-contraction+coupling+in+larval+muscles+of+Drosophila.">From action potential to contraction: Neural control and excitation-contraction coupling in larval muscles of Drosophila.</a> Comp. Biochem.Physiol. A: Molecular &#38; Integrative Physiol. 154, 173-183 (2009).</li><li>Ponzielli, R., Astier, M., Chartier, A., Gallet, A., Therond, P., Semeriva, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+tube+patterning+in+Drosophila+requires+integration+of+axial+and+segmental+information+provided+by+the+Bithorax+Complex+genes+and+hedgehog+signaling.">Heart tube patterning in Drosophila requires integration of axial and segmental information provided by the Bithorax Complex genes and hedgehog signaling.</a> Develop. 129, 4509-4521 (2002).</li><li>Sl ma, K., Farkas, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heartbeat+patterns+during+the+postembryonic+development+of+Drosophila+melanogaster.">Heartbeat patterns during the postembryonic development of Drosophila melanogaster.</a> J. Insect Physiol. 51, 489-503 (2005).</li><li>St hmer, W., Roberts, W. S., Almers, W., Sakmann, B., Neher, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Single channel recordings. The loose patch clamp, 123-132 (1983).</li><li>Wessells, R. J., Bodmer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Screening+assays+for+heart+function+mutants+in+Drosophila.">Screening assays for heart function mutants in Drosophila.</a> Biotechniques. 37, 58-66 (2004).</li><li>Zornik, E., Paisley, K., Nichols, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+transmitters+and+a+peptide+modulate+Drosophila+heart+rate.">Neural transmitters and a peptide modulate Drosophila heart rate.</a> Peptides. 20, 45-51 (1999).</li></ol>

<ol><li> Herramientas de disección: pinzas finas # 5 y tijeras finas (todos obtenidos de herramientas de Ciencias Artes (EE.UU.), Inc., 373-G Vintage Park Drive, Foster City, CA 94404 a 1.139) </li><li> Microscopio de disección con función de zoom para la disección y la tasa de conteo del corazón. Para la estimulación del centro del corazón esto es necesario también. </li><li> Para las grabaciones extracelulares e intracelulares un microscopio compuesto con los objetivos de pie (4 x y 20x) se utiliza. Amplificador estándar intracelular y A / D para el consejo sobre la grabación de la línea a una computadora. Las señales eléctricas se registran en la línea a una interfaz de PowerLab/4s (ADInstruments, Australia). Utilizamos el software estándar de ADInstruments llamado Gráfico o ámbito de aplicación. </li><li> Productos químicos: Todos los productos químicos solución salina se obtuvo de Sigma Chemical Company (St. Louis, MO). </li><li> De registros intracelulares que utilizan tubos capilares de vidrio (Catálogo # 30-31-0 de FHC, Brunswick, ME, 04011, EE.UU.) y para la grabación de macropatch focal y electrodos de estimulación se utiliza Kimax-51, Kimble productos del arte. N º 34502, ID 0.8-1.1mm, longitud 100 mm. El electrodo intracelular debe tener una resistencia de 20 a 30 Mohm. El electrodo macropatch se construye mediante la ruptura de la punta de la copa después de una punta fina se realizó a partir de un extractor de electrodos. La punta de rota tiene que ser una rotura limpia perpendicular sobre 20μM de diámetro. La punta es entonces el calor pulido de 10μM de diámetro interno para el registro extracelular focal. Para el electrodo de estimulación para conducir el corazón se utiliza una punta final de unos 20-50 m de diámetro. El eje del electrodo se ejecuta sobre un elemento de calefacción para hacer que se doble alrededor de 45 grados con una curva gradual. Esto produce un piso o una luz perpendicular electrodo sobre el tubo de corazón como el ángulo con el micromanipulador se producen alrededor de otros 45 grados a la preparación. </li></ol>

monitoring heart function in larval drosophila melanogaster for physiological studies

Se presentan varios métodos para registrar la función cardíaca en la larva<em&gt; Drosophila</em&gt;. El procedimiento permite que la frecuencia cardiaca se mide en las larvas de todo sin restricciones y restringido. Para el control directo del medio ambiente que rodea el corazón otro enfoque utiliza las larvas disecadas y la eliminación de los órganos internos con el fin de bañarse en el corazón compuestos deseados. El corazón expuesto también permite que los potenciales de membrana a controlar lo que puede dar una idea de las corrientes iónicas generadas por los miocitos y de la conducción eléctrica a lo largo del tubo cardíaco. Estos métodos tienen ventajas y desventajas para futuros experimentos que se describen. La preparación del corazón larvas ofrece un modelo adicional, además de la<em&gt; Drosophila</em&gt; Esquelético MNJ para investigar el papel de la regulación del calcio intracelular en la función celular. Aprender más acerca de las corrientes iónicas subyacentes que dan forma a los potenciales de acción en los miocitos en varias especies, que se puede esperar a tener una idea de las disfunciones iónicos conocidos asociados a genes específicos responsables de diversas enfermedades en los mamíferos.

Watch this Scientific Journal Video about Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies at JoVE.com

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

Se presenta de varias maneras para controlar la función del corazón en la larva de<em&gt; Drosophila</em&gt; Para la evaluación de preguntas relacionadas con la función de uniones, las mutaciones de los canales iónicos, la modulación de la actividad de marcapasos y los estudios farmacológicos.

Monitorización de la función del corazón en las larvas Drosophila melanogaster Para los estudios fisiológicos

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained  and  ...

monitoring-heart-function-larval-drosophila-melanogaster-for

Nanyang Technological University

Research

JoVE Journal

Biology

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

18.8K vistas.  University of Kentucky, Lexington. Se presenta de varias maneras para controlar la función del corazón en la larva de Drosophila Para la evaluación de preguntas relacionadas con la función de uniones, las mutaciones de los canales iónicos, la modulación de la actividad de marcapasos y los estudios farmacológicos.

Video: Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

Dissection of Larval CNS in Drosophila Melanogaster

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to examine the expression of various novel and marker proteins.  Staining of whole larvae is impractical because the tough cuticle prevents antibodies from penetrating inside the body cavity.  In order to stain these tissues it is necessary to dissect the animal prior to fixing and staining.  In this article we demonstrate how to dissect Drosophila larvae without damaging the CNS.  Begin by tearing the larva in half with a pair of fine forceps, and then turn the cuticle "inside-out" to expose the CNS.  If the dissection is performed carefully the CNS will remain attached to the cuticle.  We usually keep the CNS attached to the cuticle throughout the fixation and staining steps, and only completely remove the CNS from the cuticle just prior to mounting the samples on glass slides.  We also show some representative images of a larval CNS stained with Eve, a transcription factor expressed in a subset of neurons in the CNS.  The article concludes with a discussion of some of the practical uses of this technique and the potential difficulties that may arise.

In this article we demonstrate how to dissect the central nervous system from third instar Drosophila larvae.

La disección del sistema nervioso central de larvas de Drosophila melanogaster

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to ...

Investigación

Biología

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field. Studying locomotor behavior in Drosophila melanogaster relies on the ability to be able to quantify changes in motion during or in response to a given task. For this reason, a high-resolution video tracking system, such as the one we describe in this paper, is a valuable tool for measuring locomotion in real-time. Our protocol involves the use of an initial air pulse to break the flies momentum, followed by a thirty second filming period in a square chamber. A tracking program is then used to calculate the instantaneous speed of each fly within the chamber in 10 msec increments. Analysis software then compiles this data, and outputs a variety of parameters such as average speed, max speed, time spent in motion, acceleration, etc. This protocol will discuss proper feeding and management of flies for behavioral tasks, handling flies without anesthetization or immobilization, setting up a controlled environment, and running the assay from start to finish.

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement. This article demonstrates how to do this using a high-resolution tracking system.

Vídeo de alta resolución de Seguimiento de la locomoción en adultos de Drosophila melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

Drosophila Larval NMJ Immunohistochemistry

The Drosophila neuromuscular junction (NMJ) is an established model system used for the study of synaptic development and plasticity. The widespread use of the Drosophila motor system is due to its high accessibility. It can be analyzed with single-cell resolution. There are 30 muscles per hemisegment whose arrangement within the peripheral body wall are known. A total of 31 motor neurons attach to these muscles in a pattern that has high fidelity. Using molecular biology and genetics, one can create transgenic animals or mutants. Then, one can study the developmental consequences on the morphology and function of the NMJ. Immunohistochemistry can be used to clearly image the components of the NMJ. In this article, we demonstrate how to use antibody staining to visualize the Drosophila larval NMJ.

Drosophila Larval NMJ Immunohistochemistry

This protocol demonstrates how to perform immunohistochemistry on dissected Drosophila larva.

Drosophila larval NMJ inmunohistoquímica

The Drosophila neuromuscular junction (NMJ) is an established model system used for the study of synaptic development and plasticity. The widespread use ...

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.

Here we describe how to optimize the acquired video image for an olfactory magnetic-tether (OMT) apparatus. We also describe two sample experimental protocols for studying visuo-olfactory fusion.

Seguimiento visual mediada olor durante el vuelo en Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual ...

Fluorescent Labeling of Drosophila Heart Structures

The Drosophila melanogaster dorsal vessel, or heart, is a tubular structure comprised of a single layer of contractile cardiomyocytes, pericardial cells that align along each side of the heart wall, supportive alary muscles and, in adults, a layer of ventral longitudinal muscle cells. The contractile fibers house conserved constituents of the muscle cytoarchitecture including densely packed bundles of myofibrils and cytoskeletal/submembranous protein complexes, which interact with homologous components of the extracellular matrix. Here we describe a protocol for the fixation and the fluorescent labeling of particular myocardial elements from the hearts of dissected larvae and semi-intact adult Drosophila. Specifically, we demonstrate the labeling of sarcomeric F-actin and of &alpha;-actinin in larval hearts. Additionally, we perform labeling of F-actin and &alpha;-actinin in myosin-GFP expressing adult flies and of &alpha;-actinin and pericardin, a type IV extracellular matrix collagen, in wild type adult hearts. Particular attention is given to a mounting strategy for semi-intact adult hearts that minimizes handling and optimizes the opportunity for maintaining the integrity of the cardiac tubes and the associated tissues. These preparations are suitable for imaging via fluorescent and confocal microscopy. Overall, this procedure allows for careful and detailed analysis of the structural characteristics of the heart from a powerful genetically tractable model system.

Fluorescent Labeling of Drosophila Heart Structures

Here we describe a basic protocol for fluorescent labeling of different elements of heart tubes from larva and adult Drosophila melanogaster. These specimens are well-suited for imaging via fluorescent or confocal microscopy. This technique permits detailed structural analysis of the features of the hearts from a powerful model organism.

Marcaje fluorescente de Drosophila Estructuras del Corazón

The Drosophila melanogaster dorsal vessel, or heart, is a tubular structure comprised of a single layer of contractile cardiomyocytes, pericardial cells ...

Visualizing the Beating Heart in Drosophila

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in myogenic hearts. A key step in examining heart function in the fly is finding a way to access the heart in a manner that preserves its myogenic function while still allowing the beating heart organ to be observed and recorded. Two different methods for observing and recording the beating heart in both larva and adult Drosophila are described here. Our semi-intact preparation using adult flies allows clear visualization of the abdominal heart without interference from the pigmented cuticle and overlying fat bodies. To record larval heart beats it is necessary to immobilize the larva, which minimizes body wall movements thereby reducing heart movements that are not associated with myocardial contractions. Our methodologies produce stable adult and larval heart preparations that can beat for hours at rates of 1-3 Hz.

Visualizing the Beating Heart in Drosophila

Technique required for visualizing the beating heart in larval and adult Drosophila are presented. Each life stage requires a different methodology.

Visualizar el corazón latiendo en Drosophila</em

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in ...

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

The morphogenesis of the Drosophila embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell shape changes during embryonic development. One of the challenges faced in studying this structure is that the lumen of the heart tube, as well as the membrane features that are crucial to heart tube formation, are difficult to visualize in whole mount embryos, due to the small size of the heart tube and intra-lumenal space relative to the embryo. The use of transmission electron microscopy allows for higher magnification of these structures and gives the advantage of examining the embryos in cross section, which easily reveals the size and shape of the lumen. In this video, we detail the process for reliable fixation, embedding, and sectioning of late stage Drosophila embryos in order to visualize the heart tube lumen as well as important cellular structures including cell-cell junctions and the basement membrane.

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

We describe a process for fixation, embedding, sectioning, and imaging of late stage Drosophila embryos for Trasmission Electron Microscopy of the embryonic heart tube. This technique allows for the visualization of the heart tube lumen as well as the basement membrane, which lines the lumen of the heart.

Preparación de embriones para Microscopía Electrónica de la Drosophila Tubo cardíaco embrionario

The morphogenesis of the Drosophila  embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell ...

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish water and sediment. The choice of biological indicators must be based on reliable scientific knowledge and, possibly, on the availability of standardized procedures. In this article, we present a standardized protocol that used the marine crustacean Artemia to evaluate the toxicity of chemicals and/or of marine environmental matrices. Scientists propose that the brine shrimp (Artemia) is a suitable candidate for the development of a standard bioassay for worldwide utilization. A number of papers have been published on the toxic effects of various chemicals and toxicants on brine shrimp (Artemia). The major advantage of this crustacean for toxicity studies is the overall availability of the dry cysts; these can be immediately used in testing and difficult cultivation is not demanded1,2. Cyst-based toxicity assays are cheap, continuously available, simple and reliable and are thus an important answer to routine needs of toxicity screening, for industrial monitoring requirements or for regulatory purposes3. The proposed method involves the mortality as an endpoint. The numbers of survivors were counted and percentage of deaths were calculated. Larvae were considered dead if they did not exhibit any internal or external movement during several seconds of observation4. This procedure was standardized testing a reference substance (Sodium Dodecyl Sulfate); some results are reported in this work. This article accompanies a video that describes the performance of procedural toxicity testing, showing all the steps related to the protocol.

Long-term Lethal Toxicity Test with the Crustacean Artemia franciscana

This study concerns the development and standardization of a valuable methodological protocol to determine long-term (14 days) lethal toxicity exerted by chemical substances, industrial wastewater or sewage and liquid environmental samples on the saltwater crustacean, Artemia franciscana.

A largo plazo Prueba de toxicidad letal con el Crustáceo Artemia franciscana</em

Our research activities target the use of biological methods for the evaluation of environmental quality, with particular reference to saltwater/brackish ...

Measurement of Lifespan in Drosophila melanogaster

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced physical performance and increased risk of disease. Individual aging is manifest at the population level as an increase in age-dependent mortality, which is often measured in the laboratory by observing lifespan in large cohorts of age-matched individuals. Experiments that seek to quantify the extent to which genetic or environmental manipulations impact lifespan in simple model organisms have been remarkably successful for understanding the aspects of aging that are conserved across taxa and for inspiring new strategies for extending lifespan and preventing age-associated disease in mammals.
The vinegar fly, Drosophila melanogaster, is an attractive model organism for studying the mechanisms of aging due to its relatively short lifespan, convenient husbandry, and facile genetics. However, demographic measures of aging, including age-specific survival and mortality, are extraordinarily susceptible to even minor variations in experimental design and environment, and the maintenance of strict laboratory practices for the duration of aging experiments is required. These considerations, together with the need to practice careful control of genetic background, are essential for generating robust measurements. Indeed, there are many notable controversies surrounding inference from longevity experiments in yeast, worms, flies and mice that have been traced to environmental or genetic artifacts1-4. In this protocol, we describe a set of procedures that have been optimized over many years of measuring longevity in Drosophila using laboratory vials. We also describe the use of the dLife software, which was developed by our laboratory and is available for download (<a href="http://sitemaker.umich.edu/pletcherlab/software">http://sitemaker.umich.edu/pletcherlab/software</a>). dLife accelerates throughput and promotes good practices by incorporating optimal experimental design, simplifying fly handling and data collection, and standardizing data analysis. We will also discuss the many potential pitfalls in the design, collection, and interpretation of lifespan data, and we provide steps to avoid these dangers.

Measurement of Lifespan in Drosophila melanogaster

Drosophila melanogaster is a powerful model organism for exploring the molecular basis of longevity regulation. This protocol will discuss the steps involved in generating a reproducible, population-based measurement of longevity as well as potential pitfalls and how to avoid them.

Medición de la esperanza de vida en<em&gt; Drosophila melanogaster</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)

Longitudinal study of the heartbeat in small animals contributes to understanding structural and functional changes during heart development. Optical coherence microscopy (OCM) has been demonstrated to be capable of imaging small animal hearts with high spatial resolution and ultrahigh imaging speed. The high image contrast and noninvasive properties make OCM ideal for performing longitudinal studies without requiring tissue dissections or staining. Drosophila has been widely used as a model organism in cardiac developmental studies due to its high number of orthologous human disease genes, its similarity of molecular mechanisms and genetic pathways with vertebrates, its short life cycle, and its low culture cost. Here, the experimental protocols are described for the preparation of Drosophila and optical imaging of the heartbeat with a custom OCM system throughout the life cycle of the specimen. By following the steps provided in this report, transverse M-mode and 3D OCM images can be acquired to conduct longitudinal studies of the Drosophila cardiac morphology and function. The en face and axial sectional OCM images and the heart rate (HR) and cardiac activity period (CAP) histograms, were also shown to analyze the heart structural changes and to quantify the heart dynamics during Drosophila metamorphosis, combined with the videos constructed with M-mode images to trace cardiac activity intuitively. Due to the genetic similarity between Drosophila and vertebrates, longitudinal study of heart morphology and dynamics in fruit flies could help reveal the origins of human heart diseases. The protocol here would provide an effective method to perform a wide range of studies to understand the mechanisms of cardiac diseases in humans.

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy OCM

Here, the experimental protocols are described for preparing Drosophila at different developmental stages and performing longitudinal optical imaging of Drosophila heartbeats using a custom optical coherence microscopy (OCM) system. The cardiac morphological and dynamical changes can be quantitatively characterized by analyzing the heart structural and functional parameters from OCM images.

Drosophila Preparación e Imagen longitudinal de la función cardiaca in vivo utilizando microscopía de coherencia óptica (OCM)

Longitudinal study of the heartbeat in small animals contributes to understanding structural and functional changes during heart development. Optical ...

Monitorización de la función del corazón en las larvas Drosophila melanogaster Para los estudios fisiológicos

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