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. Diluições odor <ol><li> A maioria dos odores são solúveis em óleo de parafina. No entanto, DMSO ou etanol também pode ser usado como solventes alternativos para odores particular. Preparar diluições apropriadas (por exemplo, volume de 1:10: volume; v: v) de odores pura em frascos de vidro. Diluições mais odor são estáveis ​​à temperatura ambiente, mas para compostos altamente voláteis, é melhor fazer diluições trabalhando em uma base semanal. Cada sensillum responde aos odores diferentes dentro...

<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>

Pistas olfativas são usados ​​por organismos para identificar fontes de alimento, parceiros em potencial, e predadores. Neurônios sensoriais olfativos (ORS) são o centro primeiro relé entre estímulos externos e os centros superiores do cérebro onde a informação é processada. Em Drosophila melanogaster e Anopheles gambiae, ORS são facilmente acessíveis e sua atividade elétrica pode ser monitorada enquanto estimulado por puffs odor. A única técnica de grava?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<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>
</tr>
<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>
</tr>
<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

O sentido do olfato é essencial para insetos para encontrar alimentos, companheiros, predadores, e os locais de oviposição 3. Inseto neurônios sensoriais olfativos (ORS) são colocados em pêlos sensoriais chamados sensilla, que cobrem a superfície dos órgãos olfativos. A superfície de cada sensillum é coberta com minúsculos poros, através do qual odorants passar e dissolver em um líquido chamado linfa sensillum, que banha os dendritos sensorial do ORS alojados em um sensillum dado. Os dendritos OSN expressar odorant receptor (OR) proteínas, que em insetos funcionam como canais iónicos dependentes de odor 4, 5. A interação de odores com ORs aumenta ou diminui a taxa de disparo basal da OSN. Esta atividade neuronal na forma de potenciais de ação incorpora a primeira representação da qualidade, intensidade e características temporais do odorant 6, 7. Dado o fácil acesso a estes pêlos sensoriais, é possível realizar gravações extracelular de ORS único pela introdução de um eletrodo de registro para a linfa sensillum, enquanto o eletrodo de referência é colocado na linfa dos olhos ou do corpo do inseto. Em Drosophila, sensilla casa entre uma e quatro ORS, mas cada OSN normalmente exibe uma amplitude pico característico. Técnicas de classificação de Spike torná-lo possível atribuir respostas spiking a ORS individual. Este single sensillum técnica de gravação (SSR) monitora a diferença de potencial entre a linfa sensillum eo eletrodo de referência como os pontos elétricos que são gerados pela atividade do receptor de ORS 1, 2, 8. Mudanças no número de espigas, em resposta ao odor representam a base celular do odor de codificação em insetos. Aqui, descrevemos o método de preparação usado atualmente em nosso laboratório para realizar SSR em Drosophila melanogaster e Anopheles gambiae, e mostrar os traços representante induzida pela odores de forma sensillum específicos.

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

Respostas eletrofisiológicas de neurônios sensoriais olfativos de odores pode ser medido em insetos usando gravações sensillum único. Neste artigo vamos demonstrar de vídeo como realizar gravações sensillum única nas antenas da mosca do vinagre (<em&gt; Drosophila melanogaster</em&gt;) E os palpos maxilares do mosquito da malária (<em&gt; Anopheles gambiae</em&gt;).

Único Recordings Sensillum na Insetos 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

JoVE Journal

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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Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster

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When startled adult D. melanogaster react by jumping into the air and flying away. In many invertebrate species, including D. melanogaster, the "escape" ...

Visualization of Proprioceptors in Drosophila Larvae and Pupae

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In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

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Drosophila melanogaster has been used as an excellent model organism to study environmental and genetic manipulations that affect behavior. One such ...

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Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

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Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

Conditions Affecting Social Space in Drosophila melanogaster

Conditions Affecting Social Space in Drosophila melanogaster

Condições que afetam Espaço Social em<em&gt; Drosophila melanogaster</em

The social space assay described here can be used to quantify social interactions of Drosophila melanogaster &#8212; or other small insects &#8212; in a ...

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Clones em um ensaio comportamental para Mechanosensation de MarcM baseada em<em&gt; Drosophila melanogaster</em

Because of the structural and functional homology to the hair cells of the mammalian inner ear, the neurons that innervate the Drosophila external sense ...

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Uma maneira simples para medir alterações na Recompensa comportamento de busca Usando<em&gt; Drosophila melanogaster</em

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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The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

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For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...