Name: single sensillum recordings in the insects drosophila melanogaster and anopheles gambiae
Uploaded: 2010-02-17T14:25:00
Description: Watch this Scientific Journal Video about Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae at JoVE.com

1. Odore diluizioni <ol><li> La maggior parte dei odoranti sono solubili in olio di paraffina. Tuttavia, DMSO o l&#39;etanolo può essere utilizzato anche come solventi alternativi per gli odori particolari. Preparare diluizioni appropriate (ad esempio 1:10 del volume: volume, v: v) da odoranti puro in flaconcini di vetro. La maggior parte degli odori diluizioni sono stabili a temperatura ambiente, ma per i composti altamente volatili è meglio fare diluizioni di lavoro su base settimanale. Ogni sensillum risponde a d...

<ol>
	<li>Boeckh, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Elektrophysiologische+Untersuchungen+an+einzelnen+Geruchsrezeptoren+auf+der+Antenne+des+TotengrAbers+(Necrophorus+Coleoptera).">Elektrophysiologische Untersuchungen an einzelnen Geruchsrezeptoren auf der Antenne des TotengrAbers (Necrophorus Coleoptera).</a> Z. Vergl. Physiol. 46, 212-248 (1962).</li><li>Schneider, D., Hecker, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zur+Elektrophysiologic+der+Antenne+des+Seidenspinners+Bombyx+mori+bei+Reizung+mit+angereicherten+Extrakten+des+Sexuallockstoffes.">Zur Elektrophysiologic der Antenne des Seidenspinners Bombyx mori bei Reizung mit angereicherten Extrakten des Sexuallockstoffes.</a> Z. Naturforschg. 11b, 121-124 (1956).</li><li>Touhara, K., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensing+odorants+and+pheromones+with+chemosensory+receptors.">Sensing odorants and pheromones with chemosensory receptors.</a> Annu Rev Physiol. 71, 307-332 (2009).</li><li>Sato, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect+olfactory+receptors+are+heteromeric+ligand-gated+ion+channels.">Insect olfactory receptors are heteromeric ligand-gated ion channels.</a> Nature. 452, 1002-1006 (2008).</li><li>Wicher, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+odorant+receptors+are+both+ligand-gated+and+cyclic-nucleotide-activated+cation+channels.">Drosophila odorant receptors are both ligand-gated and cyclic-nucleotide-activated cation channels.</a> Nature. 452, 1007-1011 (2008).</li><li>Hallem, E. A., Ho, M. G., Carlson, J. 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+molecular+basis+of+odor+coding+in+the+Drosophila+antenna.">The molecular basis of odor coding in the Drosophila antenna.</a> Cell. 117, 965-979 (2004).</li><li>Hallem, E. A., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coding+of+odors+by+a+receptor+repertoire.">Coding of odors by a receptor repertoire.</a> Cell. 125, 143-160 (2006).</li><li>Boeckh, J., Kaissling, K. E., Schneider, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insect+olfactory+receptors.">Insect olfactory receptors.</a> Cold Spring Harb Symp Quant Biol. 30, 263-280 (1965).</li><li>de Bruyne, M., Foster, K., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor+coding+in+the+Drosophila+antenna.">Odor coding in the Drosophila antenna.</a> Neuron. 30, 537-552 (2001).</li><li>Lu, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Odor+coding+in+the+maxillary+palp+of+the+malaria+vector+mosquito+Anopheles+gambiae.">Odor coding in the maxillary palp of the malaria vector mosquito Anopheles gambiae.</a> Curr Biol. 17, 1533-1544 (2007).</li><li>Hallem, E. A., Fox, A. N., Zwiebel, L. J., Carlson, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Olfaction:+mosquito+receptor+for+human-sweat+odorant.">Olfaction: mosquito receptor for human-sweat odorant.</a> Nature. 427, 212-213 (2004).</li><li>Couto, A., Alenius, M., Dickson, B. J. M. o. l. e. c. u. l. a. r. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=anatomical,+and+functional+organization+of+the+Drosophila+olfactory+system.">anatomical, and functional organization of the Drosophila olfactory system.</a> Curr. Biol. 15, 1535-1547 (2005).</li><li>Fishilevich, E., Vosshall, L. 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+and+functional+subdivision+of+the+Drosophila+antennal+lobe.">Genetic and functional subdivision of the Drosophila antennal lobe.</a> Curr Biol. 15, 1548-1553 (2005).</li></ol>

Stimoli olfattivi vengono utilizzati dagli organismi di identificare fonti di cibo, potenziali compagni, e predatori. Neuroni sensoriali olfattivi (OSNs) sono al centro primo relè tra stimoli esterni e centri superiori del cervello dove le informazioni vengono successivamente trattati. In Drosophila melanogaster e Anopheles gambiae, OSNs sono facilmente accessibili e la loro attività elettrica possono essere monitorati, mentre stimolato da sbuffi odore. Il singolo di regi...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
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<tr>
<td>Paraffin oil</td>
<td>Odors</td>
<td>Fluka</td>
<td>76235</td>
<td>&nbsp;</td>
<tr>
<td>High purity odors (&gt;98%)</td>
<td>Odors</td>
<td>Sigma-Aldrich</td>
<td>&nbsp;</td>
<td>Methyl acetate 
#296996 
1-octen-3-ol 
#74950 </td>
</tr>
<tr>
<td>Filter paper strips</td>
<td>Odors</td>
<td>Fisherbrand</td>
<td>05-714-1</td>
<td>Chromatography paper</td>
</tr>
<tr>
<td>Connectors</td>
<td>Odors</td>
<td>Cole-Parmer</td>
<td>EW-06365-40</td>
<td>1/16x1/8"</td>
</tr>
<tr>
<td>Glass vials</td>
<td>Odors</td>
<td>Agilent Technologies</td>
<td>5182-0556</td>
<td>&nbsp;</td>
<tr>
<td>Air line plastic tubing</td>
<td>Odor Delivery</td>
<td>Python Products</td>
<td>500PAL</td>
<td>&nbsp;</td>
<tr>
<td>1 serological pipette</td>
<td>Odor Delivery</td>
<td>Corning</td>
<td>4101</td>
<td>10 mL</td>
</tr>
<tr>
<td>Plastic tubing</td>
<td>Odor Delivery</td>
<td>Cole-Parmer</td>
<td>EW-06418-0</td>
<td>0.050"x0.090"OD</td>
</tr>
<tr>
<td>Disposable borosilicate glass Pasteur pipettes</td>
<td>Odor Delivery</td>
<td>FisherBrand</td>
<td>13-678-20A</td>
<td>5-3/4 inches</td>
</tr>
<tr>
<td>Programmable stimulus controller</td>
<td>Odor Delivery</td>
<td>Syntech</td>
<td>CS-55</td>
<td>&nbsp;</td>
<tr>
<td>Anti-vibration table</td>
<td>Electrophysiology Equipment</td>
<td>TMC</td>
<td>63533</td>
<td>36&#x201D;Wx30&#x201D;Dx29&#x201D;H</td>
</tr>
<tr>
<td>Faraday cage</td>
<td>Electrophysiology Equipment</td>
<td>TMC</td>
<td>MI8133303</td>
<td>&nbsp;</td>
<tr>
<td>Inverted microscope</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>E600FN ECLIPSE</td>
<td>Recording microscope</td>
</tr>
<tr>
<td>10x and 100x objectives</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>10x Plan Fluor 100x L Plan</td>
<td>&nbsp;</td>
<tr>
<td>Dissecting microscope</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>EZ645</td>
<td>electrode sharpening/insect prep microscope</td>
</tr>
<tr>
<td>Magnetic stands</td>
<td>Electrophysiology Equipment</td>
<td>Newport</td>
<td>MODEL 150</td>
<td>&nbsp;</td>
<tr>
<td>IDAC</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>IDAC-4</td>
<td>&nbsp;</td>
<tr>
<td>Acquisition software</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>Autospike</td>
<td>&nbsp;</td>
<tr>
<td>1 macromanipulator</td>
<td>Electrophysiology Equipment</td>
<td>NARISHIGE</td>
<td>MN-151</td>
<td>Joystick manipulator 
Used for positioning reference electrode
</td>
</tr>
<tr>
<td>1 micromanipulator</td>
<td>Electrophysiology Equipment</td>
<td>EXFO</td>
<td>PCS-6000</td>
<td>Used for positioning recording electrode</td>
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<tr>
<td>Crocodile clip</td>
<td>Electrophysiology Equipment</td>
<td>Pomona</td>
<td>AL-B-12-0</td>
<td>&nbsp;</td>
<tr>
<td>Electric cable</td>
<td>Electrophysiology Equipment</td>
<td>Pomona</td>
<td>B-36-0</td>
<td>Test Cable Assembly</td>
</tr>
<tr>
<td>2 electrode holders</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>N/A</td>
<td>Electrode holders (set of 2) for tungsten wire electrode</td>
</tr>
<tr>
<td>AC probe</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>N/A</td>
<td>Universal single ended probe (10xAC)</td>
</tr>
<tr>
<td>Tungsten electrodes</td>
<td>Electrophysiology Equipment</td>
<td>Microprobes</td>
<td>M210</td>
<td>straight tungsten rods, 0.005&#x201C;x3&#x201C;</td>
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<tr>
<td>Potassium hydroxide</td>
<td>Electrophysiology Equipment</td>
<td>Sigma-Aldrich</td>
<td>221473</td>
<td>&nbsp;</td>
<tr>
<td>Syringe</td>
<td>Electrophysiology Equipment</td>
<td>BD</td>
<td>301625</td>
<td>20 mL</td>
</tr>
<tr>
<td>Power supply</td>
<td>Electrophysiology Equipment</td>
<td>WILD HEERBRUGG 6V 40W</td>
<td>e.g MTR32</td>
<td>&nbsp;</td>
<tr>
<td>Vertical puller</td>
<td>Insect prep</td>
<td>Narishige</td>
<td>PB-7</td>
<td>&nbsp;</td>
<tr>
<td>Razor blade</td>
<td>Insect prep</td>
<td>VWR</td>
<td>55411-050</td>
<td>&nbsp;</td>
<tr>
<td>Dental wax</td>
<td>Insect prep</td>
<td>Patterson</td>
<td>091-1503</td>
<td>&nbsp;</td>
<tr>
<td>Microscope slide</td>
<td>Insect prep</td>
<td>FisherBrand</td>
<td>12-550A</td>
<td>&nbsp;</td>
<tr>
<td>Cover glass</td>
<td>Insect prep</td>
<td>FisherBrand</td>
<td>12-541A</td>
<td>18X18 #1.5</td>
</tr>
<tr>
<td>Polypropylene mesh</td>
<td>Insect prep</td>
<td>Small Parts inc.</td>
<td>CMP-0500-B</td>
<td>&nbsp;</td>
<tr>
<td>Glass electrode</td>
<td>Insect prep</td>
<td>Frederick Haer & Co.</td>
<td>27-32-0-075</td>
<td>Capillary tubing borosilicate 1.5mm OD x 1.12mm ID x 75 mm</td>
</tr>
<tr>
<td>Double-sided tape (3M)</td>
<td>Insect prep</td>
<td>3M</td>
<td>MMM6652P3436</td>
<td>Double-sided tape (3M)</td>
</tr>
<tr>
<td>Forceps</td>
<td>Insect prep</td>
<td>Fine Science Tools</td>
<td>021x0053</td>
<td>Dumont #5 Mirror Finish Forceps</td>
</tr>
<tr>
<td>Small plastic cup</td>
<td>Insect prep</td>
<td>VWR</td>
<td>89009-662</td>
<td>7 x 5.7 (23/4 x 21/4)</td>
</tr>
<tr>
<td>Electric aspirator</td>
<td>Insect prep</td>
<td>Gempler&#x2019;s</td>
<td>RHM200</td>
<td>&nbsp;</td>		</tbody>
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single sensillum recordings in the insects drosophila melanogaster and anopheles gambiae

L&#39;olfatto è essenziale per gli insetti di trovare cibo, compagni, predatori e siti di deposizione delle uova 3. Insetti neuroni olfattivi sensoriali (OSNs) sono racchiusi in peli sensoriali chiamati sensilli, che coprono la superficie degli organi olfattivi. La superficie di ogni sensillum è coperta di minuscoli pori, attraverso i quali passano odoranti e si dissolvono in un liquido chiamato linfa sensillum, che bagna i dendriti sensoriali della OSNs ospitato in un sensillum dato. I dendriti OSN esprimere recettore odorizzante (OR) proteine, che in funzione di insetti come odore canali ionici 4, 5. L&#39;interazione di odoranti con OR aumenta o diminuisce la velocità basale cottura della OSN. Questa attività neuronale, sotto forma di potenziali d&#39;azione rappresenta la prima rappresentazione della qualità, intensità e caratteristiche temporali del odorizzante 6, 7. Dato il facile accesso a questi peli sensoriali, è possibile effettuare registrazioni extracellulari da OSNs singolo con l&#39;introduzione di un elettrodo di registrazione nella linfa sensillum, mentre l&#39;elettrodo di riferimento si trova nella linfa degli occhi o del corpo dell&#39;insetto. In Drosophila, sensilli casa da uno a quattro OSNs, ma ogni OSN mostra tipicamente un caratteristico picco di ampiezza. Tecniche di ordinamento Spike permettono di assegnare le risposte spiking a OSNs individuali. Questo singolo registrazione sensillum (SSR) tecnica controlla la differenza di potenziale tra la linfa sensillum e l&#39;elettrodo di riferimento come picchi elettrici che vengono generati dall&#39;attività recettore sulla OSNs 1, 2, 8. Cambiamenti nel numero di picchi in risposta alla odorizzante rappresentare la base cellulare di odore di codifica negli insetti. Qui, descriviamo il metodo di preparazione attualmente in uso nel nostro laboratorio per eseguire SSR sulla Drosophila melanogaster e Anopheles gambiae, e mostrano tracce di rappresentanza indotta dalla odoranti in un sensillum specifico modo.

Watch this Scientific Journal Video about Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae at JoVE.com

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Risposte elettrofisiologiche dei neuroni sensoriali olfattivi di odori può essere misurata in insetti utilizzando registrazioni sensillum singolo. In questo articolo il video ci mostrerà come eseguire registrazioni sensillum singolo nelle antenne del moscerino della frutta (<em&gt; Drosophila melanogaster</em&gt;) E la palpi mascellari della zanzara della malaria (<em&gt; Anopheles gambiae</em&gt;).

Registrazioni Sensillum singolo in Insetti Drosophila melanogaster E Anopheles gambiae</em

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed ...

Research

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Neuroscience

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Single-unit In vivo Recordings from the Optic Chiasm of Rat

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Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

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We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

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