Name: using single sensillum recording to detect olfactory neuron responses of bed bugs to semiochemicals
Uploaded: 2016-01-18T15:00:00
Description: Watch this Scientific Journal Video about Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals at JoVE.com

The project was supported by Award AAES 461Hatch/Multistate Grants ALA08-045 and ALA015-1-10026 to N.L.

1. Fremstilling af instrumenter, stimuli Solutions, og seng Bugs <ol><li> Der fremstilles en 50% KNO 2-opløsning (vægt / volumen) i en 20 ml flaske. </li><li> Skærpe to wolfram mikroelektroder i KNO 2-opløsning ved 5 V ved gentagne gange at dyppe wolframelektroder ind og ud af opløsningen. <ol><li> Groft skærpe wolframtråd ved at dyppe ca. 10 mm af wolfram tråd i og ud af KNO 2-opløsning ved en hastighed på 2 dips / sek i ca. 5 minutter, hvilket i høj grad kan forbruge den f...

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Det indre Sensillum Recording teknik er blevet flittigt brugt i at teste de neurale reaktioner insekter som bananfluer, myg og væggelus til forskellige kemiske stimuli i miljøet. Disse kemiske stimuli er ofte opløses og fortyndes i et fælles opløsningsmiddel for at fremstille forskellige doser af behandlinger. Dog kan forskellige opløsningsmidler producere helt andre release rater for stimuli. Tidligere undersøgelser på nogle grundigt undersøgt insekter såsom Drosophila melanogaster, malariamyg,</...

Den fælles væggelus Cimex lectularius L (Hemiptera: Cimicidae), som en midlertidig ektoparasit, er en forpligtet blodsugende insekter, hvilket betyder, deres overlevelse, udvikling og reproduktion kræver blod kilder fra værter, herunder både mennesker og dyr 1,2. Selvom virus transmission sjældent er blevet rapporteret som følge af C. lectularius, den bidende gener genereret af et angreb i alvorlig grad påvirker værter både fysisk og psykisk 3. Indførelse og udbredte anvendelse af kemiske insekticider, især DDT, sænkede risikoen for angreb, og ved udgangen af ​​1950&#39;erne angreb var på et så lavt niveau, at de ikke længere var en alvorlig bekymring i offentligheden. Imidlertid har en række mulige faktorer førte til genopblussen i bed bug befolkninger i hele verden, såsom den reducerede brug af insekticider, et fald på offentlig bevidsthed, øget rejse aktivitet, og udviklingen af resistens over for insekticider 4-9. &lt;/ p&gt; Kemiske signaler i miljøet opdages og anerkendt af insekter gennem olfaktoriske organer såsom antenner og maxillary palps. Den olfaktoriske sensilla på insekt antenner spille en afgørende rolle i at opdage disse kemiske signaler. De kemiske molekyler ind i antennal neglebånd gennem porer på neglebånd overfladen. Lugtstof bindende proteiner i antennal lymfeknuder binder til disse kemiske molekyler og transportere dem på de lugtstofreceptorer 10. De lugtstofreceptorer og deres co-receptor fra den ikke-selektive kation ionkanal på neurale membran, som vil blive depolariseret når disse kemiske molekyler er anerkendt af de lugtstofreceptorer 11. Single sensillum optagelse (SSR) blev udviklet for at detektere det ekstracellulære ændring i aktionspotentialet forårsaget af anvendelsen af ​​enten kemiske eller ikke-kemiske stimuli. Ved at indsætte en optagelse elektrode ind i sensillum lymfeknuder og en referenceelektrodei en anden del af insektets krop (som regel enten sammensatte øjne eller maven), kan afbrændingstakten af neuroner som reaktion på stimuli registreres 12. Ændringer i antallet af pigge repræsenterer følsomhed insektet til specifikke stimuli. Kemiske stimuli af forskellige identiteter og koncentration vil fremkalde forskellige neurale reaktioner med forskellige fyring satser og tidsmæssige strukturer, og kan således anvendes til at undersøge kodningsprocessen insektets for bestemte kemikalier. Til fælles bed bug begge seksuelle former deler samme mønster af olfaktoriske sensilla på antennerne: ni rillede pind C sensilla, 29 hår-lignende E (E1 og E2) sensilla, og et par enkelte Dα, Dβ, Dγ glat pind sensilla 13,14. Som flere neuroner er blevet identificeret i hver type sensillum, er det ikke let at skelne de aktionspotentialer fra forskellige neuroner anbragt i samme sensillum, så for dette eksperiment total antal aktionspotentialer blev talt off-line for en 500 ms perioden før og efter stimulering. Antallet af aktionspotentialer efter stimulering blev derefter trækkes fra antallet af aktionspotentialer før stimulering og ganget med to for at kvantificere ændringerne i fyring sats i hvert enkelt sensillum i pigge per sekund 15.

<table border="0" cellpadding="0" cellspacing="0" width="1064">
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:299px;">Tungsten wire</td>
			<td style="width:139px;">A-M SYSTEMS</td>
			<td style="width:117px;">#716500</td>
			<td style="width:511px;">Used for preparing the electrode</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">KNO2</td>
			<td>Sigma</td>
			<td>#310484</td>
			<td>Used for sharpening the tungsten wire</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">AC Power Supply</td>
			<td>BK Precision</td>
			<td>1653A</td>
			<td>Providing the voltage in sharpening the tungsten wire</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Leica Z6 APO Microscope</td>
			<td>Leica</td>
			<td>10447424</td>
			<td>Used for observing the sensilla on antennae</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Simulus controller</td>
			<td>Syntech</td>
			<td>CS-55</td>
			<td>Used for controlling the stimulus application</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">4-Channel USB Acquisition Controller</td>
			<td>Syntech</td>
			<td>IDAC-4</td>
			<td>Real-time on screen display of all signals before and during recording</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Light Source</td>
			<td>SCHOTT</td>
			<td>A20500</td>
			<td>Providing light sources for observation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Micromanupulator</td>
			<td>Leica</td>
			<td>115378</td>
			<td>Used for minor movement of electrode</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Speaker</td>
			<td>Juster</td>
			<td>95a</td>
			<td>Connected with Acquisition Controller IDAC-4 and providing sound for the signal</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Magnetic stand</td>
			<td>Narishige</td>
			<td>GJ-1</td>
			<td>Used to hold the reference electrode, stablized bed bug and stimulus delivery tube</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">TMC Vibration Isolation Table</td>
			<td>TMC</td>
			<td>63-500</td>
			<td>Used for isolating the vibration from the equipments</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Coverslip</td>
			<td>Tedpella</td>
			<td>2225-1</td>
			<td>Used for holding the bed bug</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Double-sided Tape</td>
			<td>3M</td>
			<td>XT6110</td>
			<td>Used for stablizing the bed bug on the coverclip</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Dental Wax</td>
			<td>Dentakit</td>
			<td>DK-R012</td>
			<td>Used for supporting the coverclip where bed bug is stablized&#160;</td>
		</tr>
	</tbody>
</table>

using single sensillum recording to detect olfactory neuron responses of bed bugs to semiochemicals

Insektet olfaktoriske system spiller en vigtig rolle i påvisning semiochemicals i miljøet. Især den antennal sensilla der huser enkelte eller flere neuroner inde, anses for at gøre det største bidrag med at reagere på de kemiske stimuli. Ved direkte registrering af handling potentiale i olfaktoriske sensillum efter udsættelse for stimuli, enkelt sensillum optagelse (SSR) teknik giver en kraftfuld metode til at undersøge de neurale reaktioner insekter til kemiske stimuli. Til sengen bug, som er en berygtet menneskelig parasit, har flere typer af olfaktoriske sensillum blevet karakteriseret. I denne undersøgelse, vi demonstrerede neurale reaktioner bed bug olfaktoriske sensilla til to kemiske stimuli og dosisafhængige reaktioner på en af ​​dem ved hjælp af SSR-metoden. Denne tilgang gør det muligt for forskerne at foretage tidlig screening for individuelle kemiske stimuli på sengen bug olfaktoriske sensilla, hvilket vil give værdifulde oplysninger til udvikling af nye bed bug tiltrækker eller frastøder og fordele bed bug kontrolindsats.

Enkelt sensillum optagelse er en kraftfuld efterforskningsmæssige teknik, der anvendes i studier af insekter kemisk økologi og neurale fysiologi. Undersøgelse de neurale reaktioner af insekter på forskellige flygtige forbindelser, især dem menes at være økologisk relateret til overlevelse og udvikling af insekter, ikke kun giver os uvurderlige indsigt i insekt olfaction proces, men også åbner lovende nye veje der potentielt kan medføre til udvikling af nyttige nye reagenser til...

Watch this Scientific Journal Video about Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals at JoVE.com

Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals

Bed bugs rely on olfactory receptor neurons housed in their antennal olfactory sensilla to detect semiochemicals in the environment. Utilizing single sensillum recording, we demonstrate a method to evaluate bed bug response to semiochemicals and explore the coding process involved.

Brug Single Sensillum Optagelse til Detect olfaktoriske Neuron Responses af seng Bugs til Semiochemicals

The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house ...

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