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. Odor dilutions
<ol>
<li>Most odorants are soluble in paraffin oil. However, DMSO or ethanol can also be used as alternative solvents for particular odors. Prepare appropriate dilutions (e.g. 1:10 volume : volume; v:v) from pure odorants in glass vials. Most odor dilutions are stable at room temperature, but for highly volatile compounds it is better to make working dilutions on a weekly basis. Each sensillum responds to different odors within a different concentration range. For Drosophila, a useful look-...

<ol>
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Olfactory cues are used by organisms to identify food sources, potential mates, and predators. Olfactory sensory neurons (OSNs) are the first relay center between external stimuli and higher centers of the brain where the information is further processed. In Drosophila melanogaster and Anopheles gambiae, OSNs are easily accessible and their electrical activity can be monitored while stimulated by odor puffs.
The single sensillum recording (SSR) technique explained in this vi...

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<td>Paraffin oil</td>
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<td>Fluka</td>
<td>76235</td>
<td>&nbsp;</td>
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<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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<td>Filter paper strips</td>
<td>Odors</td>
<td>Fisherbrand</td>
<td>05-714-1</td>
<td>Chromatography paper</td>
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<td>Connectors</td>
<td>Odors</td>
<td>Cole-Parmer</td>
<td>EW-06365-40</td>
<td>1/16x1/8"</td>
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<td>Glass vials</td>
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<td>Agilent Technologies</td>
<td>5182-0556</td>
<td>&nbsp;</td>
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<td>Air line plastic tubing</td>
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<td>Python Products</td>
<td>500PAL</td>
<td>&nbsp;</td>
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<td>1 serological pipette</td>
<td>Odor Delivery</td>
<td>Corning</td>
<td>4101</td>
<td>10 mL</td>
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<td>Plastic tubing</td>
<td>Odor Delivery</td>
<td>Cole-Parmer</td>
<td>EW-06418-0</td>
<td>0.050"x0.090"OD</td>
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<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>
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<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>Electrophysiology Equipment</td>
<td>Nikon</td>
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<td>10x and 100x objectives</td>
<td>Electrophysiology Equipment</td>
<td>Nikon</td>
<td>10x Plan Fluor 100x L Plan</td>
<td>&nbsp;</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>
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<td>IDAC-4</td>
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<td>Crocodile clip</td>
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<td>Pomona</td>
<td>AL-B-12-0</td>
<td>&nbsp;</td>
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<td>2 electrode holders</td>
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<td>Electrophysiology Equipment</td>
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<td>Electrophysiology Equipment</td>
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<td>Insect prep</td>
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<td>Insect prep</td>
<td>FisherBrand</td>
<td>12-541A</td>
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<td>Small Parts inc.</td>
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<td>Frederick Haer & Co.</td>
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single sensillum recordings in the insects drosophila melanogaster and anopheles gambiae

The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) proteins, which in insects function as odor-gated ion channels4, 5. The interaction of odorants with ORs either increases or decreases the basal firing rate of the OSN. This neuronal activity in the form of action potentials embodies the first representation of the quality, intensity, and temporal characteristics of the odorant6, 7.
Given the easy access to these sensory hairs, it is possible to perform extracellular recordings from single OSNs by introducing a recording electrode into the sensillum lymph, while the reference electrode is placed in the lymph of the eye or body of the insect. In Drosophila, sensilla house between one and four OSNs, but each OSN typically displays a characteristic spike amplitude. Spike sorting techniques make it possible to assign spiking responses to individual OSNs. This single sensillum recording (SSR) technique monitors the difference in potential between the sensillum lymph and the reference electrode as electrical spikes that are generated by the receptor activity on OSNs1, 2, 8. Changes in the number of spikes in response to the odorant represent the cellular basis of odor coding in insects. Here, we describe the preparation method currently used in our lab to perform SSR on Drosophila melanogaster and Anopheles gambiae, and show representative traces induced by the odorants in a sensillum-specific manner.

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

Electrophysiological responses of olfactory sensory neurons to odorants can be measured in insects using single sensillum recordings. In this video article we will demonstrate how to perform single sensillum recordings in the antennae of the vinegar fly (Drosophila melanogaster) and the maxillary palps of the malaria mosquito (Anopheles gambiae).

Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

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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