Name: identification of olfactory volatiles using gas chromatography-multi-unit recordings (gcmr) in the insect antennal lobe
Uploaded: 2013-02-24T14:00:00
Description: Watch this Scientific Journal Video about Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings GCMR in the Insect Antennal Lobe at JoVE.com

Dette arbejde blev støttet af NSF tilskud IOS 1121692, og ved University of Washington Research Foundation.

1. Flygtig Follection <ol><li> I dette eksempel anvender vi flygtige prøver fra M. lewisii blomster - en alpin wildflower indfødte til Californien. Flygtige stoffer opsamles ved hjælp af dynamiske sorption fremgangsmåder ifølge Riffell et al. 14. Kort fortalt er denne fremgangsmåde anvender et lukket kredsløb trapping system, hvor blomsterne er omgivet af en Teflon pose. Ved hjælp af en inert vakuumpumpe, er luften omkring blomsterne suges gennem en &quot;fælde&quot; består af en...

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Insekt lugte-medierede adfærd køre mange forskellige processer, herunder reproduktion, host-site markering, og identifikation af egnede føderessourcer. Undersøgelsen af ​​disse processer kræver evne til at identificere de flygtige udsendes fra kilden, såvel som evnen til at identificere de forbindelser, der medierer den adfærd. Komplicerende forhold er, at lugte består af snesevis til hundredvis af individuelle forbindelser, der tilsammen skaber en unik duft, der opfattes anderledes end de enkelte bestanddel...

<table border="1"><tbody><tr><td> Genstandens navn </td><td> Firma </td><td> Catalog Number </td><td> Kommentarer </td></tr><tr><td> Porapak Type Q 80-100 mesh </td><td> Waters </td><td> WAT027060 </td><td></td></tr><tr><td> Reynolds Ovn Tasker </td><td> Reynolds </td><td></td><td></td></tr><tr><td> GC </td><td> Agilent </td><td> 7820A </td><td></td></tr><tr><td> GC-kolonne </td><td> J &amp; W Scientific, Folsom, CA, USA </td><td> DB-5 (30 m, 0,25 mm, 0,25 um) </td><td></td></tr><tr><td> Analytisk helium bærergas </td><td> Praxair </td><td> HE K </td><td> 1 cc / min </td></tr><tr><td> 16-kanal silicium-elektrode </td><td> Neuronexus Technologies </td><td> a4x4-3mm50-177 </td><td></td></tr><tr><td> Fin tråd NiCr, 0,012 mm i diameter) </td><td> Sandvik Kanthal HP Reid </td><td> PX000004 </td><td> For at gøre brugerdefinerede tetrodes og stereotrodes </td></tr><tr><td> Forforstærker </td><td> Tucker-Davis System </td><td> PZ-2 </td><td></td></tr><tr><td> Forstærker </td><td> Tucker-Davis System </td><td> RZ-2 </td><td></td></tr><tr><td> Datafangstsystem - OpenEx suite </td><td> Tucker-Davis System </td><td></td><td></td></tr><tr><td> Online spike-sortering software - SpikePac </td><td> Tucker-Davis System </td><td></td><td></td></tr><tr><td> Offline spike-sortering software - Mclust Spike-sortering værktøjskasse </td><td> David rødlig, Institut for Neurovidenskab, Universityaf Minnesota </td><td> Gratis download på <a href="http://redishlab.neuroscience.umn.edu/MClust/MClust.html" target="_blank">http://redishlab.neuroscience.umn.edu/MClust/MClust.html</a> </td><td> MATLAB værktøjskasse </td></tr></tbody></table>

identification of olfactory volatiles using gas chromatography-multi-unit recordings (gcmr) in the insect antennal lobe

Alle organismer bebor en verden fuld af sensoriske stimuli, der bestemmer deres adfærdsmæssige og fysiologiske respons på deres omgivelser. Lugtesansen er især vigtigt i insekter, som bruger deres olfaktoriske system til at reagere på, og diskriminere blandt, komplekse lugt stimuli. Disse lugte fremkalde adfærd, som medierer processer såsom reproduktion og levesteder udvælgelse 1-3. Derudover kemiske optagelser fra insekter medierer adfærd, der har stor betydning for landbruget og menneskers sundhed, herunder bestøvning 4-6, herbivory af fødevareafgrøder 7, og overførsel af sygdomme 8,9. Identifikation af olfaktoriske signaler og deres rolle i insekt adfærd er således vigtigt for forståelsen både økologiske processer og fødevarer ressourcer og velbefindende. Hidtil har identifikationen af ​​flygtige stoffer, som driver insekt adfærd været vanskeligt og ofte kedelig. Aktuelle teknikker indbefattergaskromatografi-koblet electroantennogram optagelse (GC-EAG), og gaskromatografi-koblede enkelt sensillum optagelser (GC-SSR) 10-12. Disse teknikker viste sig at være afgørende i identifikationen af ​​bioaktive stoffer. Vi har udviklet en metode, der benytter gaschromatografi koblet til multi-kanal elektrofysiologiske optagelser (benævnt »GCMR«) fra neuroner i antennal lap (AL, insektet primære olfaktoriske center) 13,14. Denne state-of-the-art teknik giver os mulighed for at undersøge hvordan lugt information er repræsenteret i insektet hjernen. Endvidere, fordi neurale reaktioner på lugte på dette niveau af olfaktoriske behandling er meget følsomme på grund af graden af ​​konvergens af antennens receptor neuroner i AL neuroner, vil AL-optagelser tillade påvisningen af ​​aktive bestanddele i naturlige lugte effektivt og med høj følsomhed. Her beskriver vi GCMR og give et eksempel på dets anvendelse. Adskillige generelle trin er involat the i afsløring af bioaktive flygtige stoffer og insekt respons. Flygtige først skal indsamles fra kilder af interesse (i dette eksempel bruger vi blomster fra slægten Mimulus (Phyrmaceae)) og karakteriseres efter behov ved hjælp af standard GC-MS-metoder 14-16. Insekter er forberedt til studie med minimal dissektion, hvorefter en optagelse elektrode sættes ind i antennal lap og multi-kanal neural optagelse begynder. Post-behandling af de neurale data derefter afslører, hvilke specifikke lugtstoffer medføre betydelige neurale reaktioner fra insektet nervesystemet. Selvom eksemplet præsenterer vi her er specifikt for bestøvning undersøgelser, kan GCMR udvides til en lang række studier organismer og flygtige kilder. For eksempel kan denne fremgangsmåde anvendes til identifikation af lugtstoffer der tiltrækker eller frastøder insekter og skadedyr i afgrøden. Desuden kan GCMR også anvendes til at identificere tiltrækningsmidler for gavnlige insekter såsom pollinators. Teknikken kan udvides til ikke-insektceller emner også.

I GCMR assay under anvendelse af M. lewisii blomstret duft, vi injicere 3 pi af ekstraktet i GC. Det samlede antal flygtige stoffer eluerede ved GC er typisk 60-70 flygtige stoffer. Duften af M. lewisii er overvejende sammensat af monoterpenoids, herunder β-myrcen (acykliske) og α-pinen, med den resterende del af duft sammensat af seks carbon-flygtige stoffer, såsom 2-hexanol, og sesquiterpenoids der omfatter &lt;1% af frirummet. GCMR udnytter følsomhe...

Watch this Scientific Journal Video about Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings GCMR in the Insect Antennal Lobe at JoVE.com

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings GCMR in the Insect Antennal Lobe

Olfaktoriske stikord medierer mange forskellige adfærd hos insekter, og er ofte komplekse blandinger bestående af snesevis til hundredvis af flygtige forbindelser. Anvendelse af gaskromatografi med flere kanaler optagelse i insekt antennal lap, beskriver vi en fremgangsmåde til identifikation af bioaktive forbindelser.

Identifikation af olfaktoriske Flygtige stoffer ved hjælp af gaskromatografi-Multi-unit Recordings (GCMR) i Insect Antennal Lobe

All  organisms inhabit a world full of sensory stimuli that determine their  behavioral and physiological response to their environment. Olfaction is ...

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Neuroscience

Identification of Olfactory Volatiles using Gas Chromatography-Multi-unit Recordings (GCMR) in the Insect Antennal Lobe

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe

Calcium Imaging af lugt-fremkaldte reaktioner i Drosophila Antennal Lobe

The antennal lobe is the primary olfactory center in the insect brain and represents the anatomical and functional equivalent of the vertebrate olfactory ...

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Dobbelte Elektrofysiologiske Optagelser af synaptisk-evoked astrogliale og Neuronal respons i Akut hippocampusskiver

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

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Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Using Insect Electroantennogram Sensors on Autonomous Robots for Olfactory Searches

Brug Insekt Electroantennogram Sensorer på selvstyrende robotter til Olfaktoriske søgninger

Robots designed to track chemical leaks in hazardous industrial facilities1 or explosive traces in landmine fields2 face the same problem as insects ...

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Ex vivo Tilberedninger af den intakte Vomeronasal orgel og tilbehør lugtekolben

The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first ...

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Bestemmelse af Lysmodtageren Cell spektral følsomhed en Insect Model fra<em&gt; In vivo</em&gt; Intracellulære optagelser

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Quadruple Immunostaining of the Olfactory Bulb for Visualization of Olfactory Sensory Axon Molecular Identity Codes

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The mouse olfactory system is often used to study mechanisms of neural circuit formation&#12288;because of its simple anatomical structure. An Olfactory ...