Name: obtaining hemocytes from the hawaiian bobtail squid euprymna scolopes and observing their adherence to symbiotic and non-symbiotic bacteria
Uploaded: 2010-02-11T04:52:00
Description: Watch this Scientific Journal Video about Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria at JoVE.com

Fontes de financiamento: Universidade de Connecticut Research Foundation e do Departamento de Biologia Molecular e Celular para SVN, Sigma Xi Grant-in-Aid de Pesquisa e H. e R. Antonio Majorie Romano de Pós-Graduação Educação Fellowship para AJC

<ol><li> Prepare 500 mL de 0,22 M esterilizada por filtração, a água do mar artificial (FSW; salinidade 35 ppt). Filtro de água do mar artificial ou natural através de um filtro de 0,22 micron M para remover partículas e bactérias. </li><li> Anestesiar um adulto Hawaiian bobtail squid (Euprymna scolopes), colocando em uma solução 2% de etanol na FSW. Colocar o animal na anestesia por aproximadamente 10 minutos. A lula deixará de natação e não responder ativamente ao toque. Respiração contínua, indicado pelo movimento do...

<ol>
	<li>Koropatnick, T. A., Kimbell, J. R., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responses+of+host+hemocytes+during+the+initiation+of+the+squid-Vibrio+symbiosis.">Responses of host hemocytes during the initiation of the squid-Vibrio symbiosis.</a> Biol. Bull. 212, 29-39 (2007).</li><li>McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unseen+forces:+the+influence+of+bacteria+on+animal+development.">Unseen forces: the influence of bacteria on animal development.</a> Dev. Biol. 242, 1-14 (2002).</li><li>McFall-Ngai, M. J., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Symbiont+recognition+and+subsequent+morphogenesis+as+early+events+in+an+animal-bacterial+mutualism.">Symbiont recognition and subsequent morphogenesis as early events in an animal-bacterial mutualism.</a> Science. 254, 1491-1494 (1991).</li><li>Nyholm, S. V., Stewart, J. J., Ruby, E. G., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recognition+between+symbiotic+Vibrio+fischeri+and+the+haemocytes+of+Euprymna+scolopes.">Recognition between symbiotic Vibrio fischeri and the haemocytes of Euprymna scolopes.</a> Environ. Microbiol. 11, 483-493 (2009).</li><li>Nyholm, S. V., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+winnowing:+establishing+the+squid-Vibrio+symbiosis.">The winnowing: establishing the squid-Vibrio symbiosis.</a> Nat. Rev. Microbiol. 2, 632-642 (2004).</li><li>Nyholm, S. V., Stabb, E. V., Ruby, E. G., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+of+an+animal-bacterial+association:+recruiting+symbiotic+Vibrios+from+the+environment.">Establishment of an animal-bacterial association: recruiting symbiotic Vibrios from the environment.</a> Proc. Natl. Acad. Sci. USA. 97, 10231-10235 (2000).</li><li>Nyholm, S. V., McFall-Ngai, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sampling+the+light-organ+microenvironment+of+Euprymna+scolopes:+description+of+a+population+of+host+cells+in+association+with+the+bacterial+symbiont+Vibrio+fischeri.">Sampling the light-organ microenvironment of Euprymna scolopes: description of a population of host cells in association with the bacterial symbiont Vibrio fischeri.</a> Biol Bull. 195, 89-97 (1998).</li><li>Visick, K. L., Ruby, E. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vibrio+fischeri+and+its+host:+it+takes+two+to+tango.">Vibrio fischeri and its host: it takes two to tango.</a> Curr. Opin. Microbiol. 9, 632-638 (2006).</li></ol>

Estudos sobre o papel do sistema imunológico na mediação de sinalização molecular entre bactérias benéficas e os seus anfitriões têm, nos últimos anos, fez contribuições significativas para a nossa compreensão da co-evolução dos eucariotos com sua microbiota. O sistema squid / vibrio provou-se como um sistema modelo tratável para responder a perguntas fundamentais neste campo 2,3,5,6,8. A luz de órgãos do Euprymna squid scolopes permite a colonização exclusivamente pela b...

<!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>Lab-Tek chambered #1.0 borosilicate coverglass system (8-chambers)</td>
<td>&nbsp;</td>
<td>Thermo-Fisher Scientific</td>
<td>155411</td>
<td>&nbsp;</td>
<tr>
<td>26.5 G 1ml latex-free insulin syringe</td>
<td>&nbsp;</td>
<td>Becton, Dickinson and Company</td>
<td>C34551</td>
<td>&nbsp;</td>
<tr>
<td>Cell Tracker Orange</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>C34551</td>
<td>&nbsp;</td>
<tr>
<td>SteREO Discovery V20 Microscope</td>
<td>&nbsp;</td>
<td>Zeiss</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>SP2 Confocal Microscope</td>
<td>&nbsp;</td>
<td>Leica</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

obtaining hemocytes from the hawaiian bobtail squid euprymna scolopes and observing their adherence to symbiotic and non-symbiotic bacteria

Estudos sobre o papel do sistema imunológico na mediação de sinalização molecular entre bactérias benéficas e os seus anfitriões têm, nos últimos anos, fez contribuições significativas para a nossa compreensão da co-evolução dos eucariotos com sua microbiota. A associação simbiótica entre o havaiano bobtail lula,<em&gt; Euprymna scolopes</em&gt; E bactéria bioluminescente<em&gt; Vibrio fischeri</em&gt; Tem sido utilizado como um sistema modelo para entender os efeitos das bactérias benéficas no desenvolvimento animal. Estudos recentes têm demonstrado que macrófagos-like hemócitos, o único componente celular do sistema inato do hospedeiro lula imune, provavelmente desempenham um papel importante na mediação do estabelecimento e manutenção da associação. Este protocolo irá demonstrar como obter hemócitos de<em&gt; E. scolopes</em&gt; E depois usar essas células em ensaios de ligação bacteriana. Adulto lula são os primeiros anestesiados antes de hemolinfa coletadas pela seringa do vaso sanguíneo principal cefálica. Os hemócitos host, contidas na hemolinfa extraídos, são aderidas em lamínulas de vidro câmaras e expostos à proteína fluorescente verde marcado simbiótica<em&gt; Vibrio fischeri</em&gt; E não-simbiótica<em&gt; Vibrio harveyi</em&gt;. Os hemócitos são contrastado com um corante fluorescente (Cell Rastreador de Orange, Invitrogen) e, em seguida, visualizado por microscopia de fluorescência.

Watch this Scientific Journal Video about Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria at JoVE.com

Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria

Este vídeo vai demonstrar como obter hemócitos (células sangüíneas) a partir do Hawaiian bobtail lula,<em&gt; Euprymna scolopes</em&gt; Para utilização em ensaios biológicos de células bacterianas e aderência. Hemócitos serão marcadas com um corante fluorescente e expostos a GFP-rotulados bactérias.

Obtenção Hemócitos do Hawaiian Bobtail Squid Euprymna scolopes Observando e sua adesão a bactérias simbióticas e não-simbióticas

Studies concerning the role of the immune system in mediating molecular signaling between beneficial bacteria and their hosts have, in recent years, made ...

Research

JoVE Journal

Biology

Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria

Obtaining Eggs from Xenopus laevis Females

Obtenção de ovos de Xenopus laevis fêmeas

The eggs of Xenopus laevis intact, lysed, and/or fractionated are useful for a wide variety of experiments. This protocol shows how to induce egg laying, ...

Biologia

Obtaining High Quality RNA from Single Cell Populations in Human Postmortem Brain Tissue

Obtenção de RNA de alta qualidade a partir de populações de células no tecido humano único post-mortem do cérebro

We proposed to investigate the gray matter reduction in the superior temporal gyrus seen in schizophrenia patients, by interrogating gene expression ...

Formulations for Freeze-drying of Bacteria and Their Influence on Cell Survival

As formulações para liofilização de bactérias e sua influência na sobrevivência das células

Cellular water can be removed to reversibly inactivate microorganisms to facilitate storage. One such method of removal is freeze-drying, which is ...

A Simple Protocol for Extracting Hemocytes from Wild Caterpillars

Um protocolo simples para extrair Hemócitos de Lagartas selvagens

Insect hemocytes (equivalent to mammalian white blood cells) play an important role in several physiological processes throughout an insect's life cycle ...

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

Isolamento e Cultura de miofibras individuais e suas células satélites do músculo esquelético

Muscle  regeneration in the adult is performed by resident stem cells called satellite  cells. Satellite cells are defined by  their position between the ...

Collection, Isolation and Enrichment of Naturally Occurring Magnetotactic Bacteria from the Environment

Isolamento, coleta e Enriquecimento de bactérias naturais magnetotáticas do Meio Ambiente

Magnetotactic bacteria (MTB) are aquatic microorganisms that were first notably described in 19751 from sediment samples collected in salt marshes of ...

Obtaining Specimens with Slowed, Accelerated and Reversed Aging in the Honey Bee Model

Obtenção de amostras com retardado, acelerado e invertida Envelhecimento no Modelo Bee Mel

Societies of highly social animals feature vast lifespan differences between closely related individuals. Among social insects, the honey bee is the best ...

A Method for Extracting Pigments from Squid Doryteuthis pealeii

A Method for Extracting Pigments from Squid Doryteuthis pealeii

Um método para extrair Pigmentos de Squid<em&gt; Pealeii Doryteuthis</em

Cephalopods can undergo rapid and adaptive changes in dermal coloration for sensing, communication, defense, and reproduction purposes. These capabilities ...

Split Green Fluorescent Protein System to Visualize Effectors Delivered from Bacteria During Infection

Sistema de proteína verde fluorescente Split Visualizar efetores entregue de bactérias durante a infecção

Bacteria, one of the most important causative agents of various plant diseases, secrete a set of effector proteins into the host plant cell to subvert the ...

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current

Isolation of High Quality Murine Atrial and Ventricular Myocytes for Simultaneous Measurements of Ca2+ Transients and L-Type Calcium Current

Isolamento de miócitos murinos atrial e ventricular de alta qualidade para medidas simultâneas de Ca2+ transitórios e corrente de cálcio tipo L

Mouse models play a crucial role in arrhythmia research and allow studying key mechanisms of arrhythmogenesis including altered ion channel function and ...