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

Fonti di finanziamento: University of Connecticut Research Foundation e dal Dipartimento di biologia molecolare e cellulare a SVN, Sigma Xi Grant-in-Aid-di Ricerca e Antonio H. e R. Majorie Romano Graduate Education Fellowship per AJC

<ol><li> Preparare 500 ml di 0,22 micron, sterilizzata per filtrazione di acqua di mare artificiale (FSW; salinità 35 ppt). Filtro acqua di mare artificiale o naturale attraverso un micron 0,22 micron filtro per rimuovere le particelle e batteri. </li><li> Anestetizzare un adulto hawaiano bobtail calamari (Euprymna scolopes), mettendo in una soluzione al 2% di etanolo in FSW. Posto l&#39;animale in anestesia per circa 10 minuti. Il calamaro cesserà di nuoto e non rispondere attivamente al tatto. Respirazione continua, indicato dal mo...

<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>

Studi riguardanti il ​​ruolo del sistema immunitario nella mediazione di segnale molecolare tra batteri benefici ei loro ospiti hanno, negli ultimi anni, dato un contributo significativo alla nostra comprensione della co-evoluzione degli eucarioti, con la loro flora batterica. Il calamaro / vibrione del sistema si è dimostrato come sistema modello trattabile per rispondere alle domande fondamentali in questo campo 2,3,5,6,8. La luce-organo del Euprymna calamari scolopes permette di colo...

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<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>
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<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>
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<td>Cell Tracker Orange</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>C34551</td>
<td>&nbsp;</td>
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<td>SteREO Discovery V20 Microscope</td>
<td>&nbsp;</td>
<td>Zeiss</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>SP2 Confocal Microscope</td>
<td>&nbsp;</td>
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obtaining hemocytes from the hawaiian bobtail squid euprymna scolopes and observing their adherence to symbiotic and non-symbiotic bacteria

Studi riguardanti il ​​ruolo del sistema immunitario nella mediazione di segnale molecolare tra batteri benefici ei loro ospiti hanno, negli ultimi anni, dato un contributo significativo alla nostra comprensione della co-evoluzione degli eucarioti, con la loro flora batterica. L&#39;associazione simbiotica tra l&#39;hawaiano bobtail calamari,<em&gt; Euprymna scolopes</em&gt; E il batterio bioluminescente<em&gt; Vibrio fischeri</em&gt; È stato utilizzato come sistema modello per la comprensione degli effetti di batteri benefici sullo sviluppo animale. Recenti studi hanno dimostrato che macrofagi come ematociti concentrati, il componente esclusivo cellulare del sistema immunitario dell&#39;ospite calamaro, probabilmente svolgono un ruolo importante nel mediare la creazione e il mantenimento di questa associazione. Questo protocollo dimostrerà come ottenere ematociti concentrati da<em&gt; E. scolopes</em&gt; E poi usare queste cellule batteriche in saggi di binding. Adulti calamari vengono prima anestetizzati prima di emolinfa viene raccolta da siringa dal vaso sanguigno principale cefalica. Il ematociti concentrati host, contenuta nel emolinfa estratte, sono rispettati coprioggetto di vetro a camera e poi esposto alla proteina verde fluorescente etichettati simbiotico<em&gt; Vibrio fischeri</em&gt; E non-simbiotico<em&gt; Vibrio harveyi</em&gt;. Il ematociti concentrati sono di contrasto con un colorante fluorescente (Cell Tracker Orange, Invitrogen) e poi visualizzate utilizzando la microscopia a fluorescenza.

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

Questo video dimostra come ottenere ematociti concentrati (cellule del sangue) dal hawaiano bobtail calamari,<em&gt; Euprymna scolopes</em&gt; Per l&#39;utilizzo in saggi di adesione cellulare biologico e batteriche. Ematociti concentrati sarà tinto con un colorante fluorescente ed esposto a GFP-etichettata batteri.

Ottenere ematociti concentrati dal hawaiano Bobtail Squid Euprymna scolopes L&#39;osservazione e la loro aderenza ai batteri simbionti e non Symbiotic

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

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

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