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. Geur verdunningen <ol><li> De meeste geurstoffen zijn oplosbaar in paraffine olie. Echter, DMSO of ethanol ook worden gebruikt als alternatief oplosmiddel voor bepaalde geuren. Bereid geschikte verdunningen (bijv. 1:10 volume: volume, v: v) uit zuivere geurstoffen in glazen flesjes. De meeste geur verdunningen zijn stabiel bij kamertemperatuur, maar voor zeer vluchtige stoffen is het beter te werken verdunningen te maken op een wekelijkse basis. Elke sensillum reageert op de verschillende geuren in een andere conce...

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Olfactorische signalen worden gebruikt door organismen aan voedsel bronnen, mogelijke partners, en roofdieren te identificeren. Olfactorische sensorische neuronen (OSNs) zijn de eerste relais midden tussen externe stimuli en de hogere centra van de hersenen waar de informatie verder wordt verwerkt. In Drosophila melanogaster en Anopheles gambiae, OSNs zijn gemakkelijk bereikbaar en hun elektrische activiteit kan worden gecontroleerd, terwijl gestimuleerd door geur trekjes. ...

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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>
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<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>
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<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>
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<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>
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<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>
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<td>Faraday cage</td>
<td>Electrophysiology Equipment</td>
<td>TMC</td>
<td>MI8133303</td>
<td>&nbsp;</td>
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<td>Inverted microscope</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
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<td>10x and 100x objectives</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
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<td>Dissecting microscope</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>EZ645</td>
<td>electrode sharpening/insect prep microscope</td>
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<td>Magnetic stands</td>
<td>Electrophysiology Equipment</td>
<td>Newport</td>
<td>MODEL 150</td>
<td>&nbsp;</td>
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<td>IDAC</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>IDAC-4</td>
<td>&nbsp;</td>
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<td>Acquisition software</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
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<td>Joystick manipulator 
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<td>1 micromanipulator</td>
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<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>
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<td>Electric cable</td>
<td>Electrophysiology Equipment</td>
<td>Pomona</td>
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<td>2 electrode holders</td>
<td>Electrophysiology Equipment</td>
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<td>N/A</td>
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<td>AC probe</td>
<td>Electrophysiology Equipment</td>
<td>Syntech</td>
<td>N/A</td>
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<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>
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<td>221473</td>
<td>&nbsp;</td>
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<td>Syringe</td>
<td>Electrophysiology Equipment</td>
<td>BD</td>
<td>301625</td>
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<td>Power supply</td>
<td>Electrophysiology Equipment</td>
<td>WILD HEERBRUGG 6V 40W</td>
<td>e.g MTR32</td>
<td>&nbsp;</td>
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<td>Vertical puller</td>
<td>Insect prep</td>
<td>Narishige</td>
<td>PB-7</td>
<td>&nbsp;</td>
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<td>Razor blade</td>
<td>Insect prep</td>
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<td>55411-050</td>
<td>&nbsp;</td>
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<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>
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<tr>
<td>Forceps</td>
<td>Insect prep</td>
<td>Fine Science Tools</td>
<td>021x0053</td>
<td>Dumont #5 Mirror Finish Forceps</td>
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<tr>
<td>Small plastic cup</td>
<td>Insect prep</td>
<td>VWR</td>
<td>89009-662</td>
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<td>Electric aspirator</td>
<td>Insect prep</td>
<td>Gempler&#x2019;s</td>
<td>RHM200</td>
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single sensillum recordings in the insects drosophila melanogaster and anopheles gambiae

Het gevoel van geur is van essentieel belang voor insecten te vinden voedsel, partners, roofdieren, en ovipositie plaatsen 3. Insect olfactorische sensorische neuronen (OSNs) zijn ingesloten in de sensorische haren genaamd sensilla, die het oppervlak van de reukorganen te dekken. Het oppervlak van elk sensillum is bedekt met kleine poriën, waardoor geurstoffen passeren en oplossen in een vloeistof genaamd lymfe sensillum, die de sensorische dendrieten van de OSNs gehuisvest in een bepaalde sensillum baadt. De OSN dendrieten express geurstof receptor (OR) eiwitten, die bij insecten functioneren als geur-gated ionkanalen 4, 5. De interactie van geurstoffen met de OR of stijgingen of dalingen van de basale vuursnelheid van de OSN. Deze neuronale activiteit in de vorm van actiepotentialen belichaamt de eerste voorstelling van de kwaliteit, intensiteit, en temporele kenmerken van de geurstof 6, 7. Gezien de gemakkelijke toegang tot deze zintuiglijke haren, is het mogelijk om extracellulaire opnames van enkele OSNs uit te voeren door de invoering van een registratie-elektrode in de sensillum lymfe, terwijl de referentie-elektrode wordt geplaatst in de lymfe van het oog of het lichaam van het insect. In Drosophila, sensilla huis tussen een en vier OSNs, maar elke OSN geeft meestal een karakteristieke piek amplitude. Spike sorteer-technieken maken het mogelijk om stekelige reacties toewijzen aan de individuele OSNs. Deze single sensillum opname (SSR) techniek bewaakt het verschil in potentiaal tussen de sensillum lymfe en de referentie-elektrode als elektrische pieken die worden gegenereerd door de receptor activiteit op OSNs 1, 2, 8. Veranderingen in het aantal pieken in reactie op de geurstof vertegenwoordigen de cellulaire basis van geur codering bij insecten. We beschrijven hier de bereidingswijze die momenteel worden gebruikt in ons lab te SSR te voeren op Drosophila melanogaster en Anopheles gambiae, en laten zien vertegenwoordiger sporen geïnduceerd door de geurstoffen in een sensillum-specifieke wijze.

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

Elektrofysiologische reacties van olfactorische sensorische neuronen aan geurstoffen kan worden gemeten bij insecten met behulp van enkele sensillum opnames. In deze video artikel zullen we laten zien hoe je een sensillum opnamen in de antennes van de azijn fly uit te voeren (<em&gt; Drosophila melanogaster</em&gt;) En de maxillaire palpen van de malariamug (<em&gt; Anopheles gambiae</em&gt;).

Single Sensillum Opnamen in de Insecten Drosophila melanogaster En 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 ...

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Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

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