Name: neuropharmacological manipulation of restrained and free-flying honey bees, apis mellifera
Uploaded: 2016-11-26T13:00:00
Description: Watch this Scientific Journal Video about Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera at JoVE.com

This project was funded by ARC grant DP0986021 and NHMRC grant 585442. ABB is supported by an ARC Future Fellowship (FT140100452). JAP is supported by an iMQRES scholarship awarded by Macquarie University and by a DAAD-Doktorandenstipendium awarded by the German Academic Exchange Service. JMD is supported by CNRS and University Paul Sabatier.

1. Drug Administration for utnyttet Bees <ol><li> oral behandling <ol><li> Fremstille 1,5 M sukrose-løsning ved å blande 257 g sukrose med 500 ml vann (det er lettere å oppløse denne mengden av sukrose i kokende vann). Oppbevar sukroseoppløsning ved 4 ° C inntil bruk. MERK: Sukrose løsningen gir en meget gjestfritt miljø for visse mikroorganismer, og dermed lett blir forurenset og usmakelig for bier. Bulk sukroseløsning kan alikvotert og lagret ved -20 ° C inntil bruk. </li><li> Avgj?...

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Metodene som er skissert ovenfor tillate enkel, effektiv og robust behandling av enten frittflygende eller temte honningbier. Disse metodene er kompatibel med mange eksperimentelle paradigmer og biologiske spørsmål (tabell 1). Alle de frittflygende metoder kan lett brukes til å utnyttes bier. Det motsatte er mindre vellykket, men siden midlertidig tilbakeholdenhet og invasive behandlingsmetoder kan ofte inngå kompromisser bienes flying evne. Metodene har blitt presentert...

Siden Karl von Frisch belyst deres dansespråk 1, har honningbier vært et populært studie arter for forskere i dyrs atferd og nevrobiologi. I de senere år har en myriade av nye disipliner har dukket opp i skjæringspunktet mellom disse to feltene, og flere andre disipliner (f.eks, molekylærbiologi, genomikk og informatikk) har oppstått sammen med dem. Dette har ført til rask utvikling av nye teorier og modeller for å forstå hvordan atferd skyldes aktivitet i nervesystemet. På grunn av den unike livsstil, rik atferdsrepertoar, og enkel eksperimentell og farmakologisk manipulasjon, har bier holdt i forkant av denne revolusjonen. Honningbier blir brukt til å studere grunnleggende nevrobiologiske spørsmål som de underliggende læring og hukommelse 2,3, beslutnings 4, olfactory 5, eller visuell prosessering 6. I de siste årene har honey bee har også blitt brukt som modell for å studere emner som regel reservert for medisinsk forskning, for eksempel effekten av vanedannende legemidler 7 - 11, sove 12, aldring 13, eller mekanismene som ligger til grunn anestesi 14. I motsetning til de klassiske genetisk modellorganismer (f.eks D. melanogaster, C. elegans, M. musculus), det er svært få genetiske verktøy for å manipulere nevrale funksjoner i honningbier, selv om dette er i ferd med å endre 15. I stedet har honningbie studier primært avhengig av farmakologiske manipulasjoner. Dette har vært svært vellykket; derimot, er mangfoldet av bee forskning slik at en rekke metoder for farmakologisk administrasjon er nødvendig. Forskning med honningbier løser svært ulike spørsmål, er studert av forskere fra ulike disipliner og bakgrunner, og bruker en rekke eksperimentelle tilnærminger. mange Resebue spørsmål krever bier til enten være frittflygende, fritt samspill i sin koloni, eller begge deler. Dette kan gjøre det vanskelig å holde oversikt over individuelle forsøksdyr, og gjør beherskelse eller kanylering unfeasible. For å imøtekomme mangfoldet av honningbie forskning, er en rekke av stoffet leveringsmetoder som trengs, slik at for robust og fleksibel administrasjon og samtidig sikre at den farmakokinetiske og farmakodynamiske profiler, invasivitet av metoden, og dens pålitelighet, passer paradigmet i spørsmålet. På grunn av disse ulike behov, har de fleste forskningsmiljøer utviklet sine egne unike Drug Administration metoder. Så langt har dette vært en styrke på bee forskersamfunnet; det har ført til utviklingen av matriser av metoder som åpner for bruk av det samme stoffet i ulike situasjoner. Vårt mål her er ikke å utvikle et enkelt standardisert metode for farmakologiske manipulasjoner av bier, men heller å fremheve metoder somhar vist seg å være spesielt vellykket, og hjelpe forskere vedta disse. Vi diskuterer de grunnleggende prinsippene for hvordan de fungerer, så vel som sine fordeler og ulemper.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Sucrose</td>
			<td>Sigma-Aldrich</td>
			<td>S8501</td>
			<td>Any supplier will do</td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Sigma-Aldrich</td>
			<td>S7653</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Chloride</td>
			<td>Sigma-Aldrich</td>
			<td>P9333&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium Chloride hexahydrate</td>
			<td>Sigma-Aldrich</td>
			<td>M2670</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium Chloride dihydrate</td>
			<td>Sigma-Aldrich</td>
			<td>C8106</td>
			<td></td>
		</tr>
		<tr>
			<td>Dextrose monohydrate</td>
			<td>Sigma-Aldrich</td>
			<td>49159</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffer Saline (PBS)</td>
			<td>Sigma-Aldrich</td>
			<td>P4417</td>
			<td></td>
		</tr>
		<tr>
			<td>Protection Wax</td>
			<td>Dentaurum</td>
			<td>124-305-00</td>
			<td></td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>Sigma-Aldrich</td>
			<td>H3375</td>
			<td></td>
		</tr>
		<tr>
			<td>dimethylformamide</td>
			<td>Sigma-Aldrich</td>
			<td>D4551</td>
			<td></td>
		</tr>
		<tr>
			<td>95% Ethanol</td>
			<td>Sigma-Aldrich</td>
			<td>493511</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass capillary</td>
			<td>WPI</td>
			<td>1B100F-3</td>
			<td></td>
		</tr>
		<tr>
			<td>23 G NanoFil needle</td>
			<td>WPI</td>
			<td>NF33BV-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Very fine forsceps</td>
			<td>Dumont</td>
			<td>0208-55-PO</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode puller</td>
			<td>SRI</td>
			<td>2001</td>
			<td></td>
		</tr>
		<tr>
			<td>FemtoJet Microinjector</td>
			<td>Eppendorf</td>
			<td>5247 000.01</td>
			<td></td>
		</tr>
		<tr>
			<td>Eicosane</td>
			<td>Sigma-Aldrich</td>
			<td>219274</td>
			<td></td>
		</tr>
		<tr>
			<td>manual micromanipulator</td>
			<td>Brinkmann Instrumentenbau</td>
			<td>MM-33</td>
			<td></td>
		</tr>
		<tr>
			<td>electronic micromanipulator</td>
			<td>Luigs &#38; Neumann Feinmechanik + Elektortechnik</td>
			<td>Junior unit XYZ</td>
			<td></td>
		</tr>
		<tr>
			<td>stereomicroscope</td>
			<td>Leica</td>
			<td>M80</td>
			<td></td>
		</tr>
		<tr>
			<td>soldering iron</td>
			<td>Weller</td>
			<td>WESD51</td>
			<td></td>
		</tr>
		<tr>
			<td>Dextran, Alexa Fluor 546, 10000 MW</td>
			<td>ThermoFisher Scientific</td>
			<td>D-22911</td>
			<td></td>
		</tr>
		<tr>
			<td>Dextran, Alexa Fluor 568, 10000 MW</td>
			<td>ThermoFisher Scientific</td>
			<td>D-22912</td>
			<td></td>
		</tr>
		<tr>
			<td>small Petri dish</td>
			<td>Sigma-Aldrich</td>
			<td>P5481</td>
			<td></td>
		</tr>
		<tr>
			<td>mineral oil</td>
			<td>Sigma-Aldrich</td>
			<td>M5904</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL Centrifuge tube</td>
			<td>ThermoFisher Scientific</td>
			<td>339652</td>
			<td></td>
		</tr>
		<tr>
			<td>forceps</td>
			<td>Australian Entomological Supplies</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Blade holder and breaker</td>
			<td>Australian Entomological Supplies</td>
			<td>E130</td>
			<td></td>
		</tr>
		<tr>
			<td>Feather double edged razor blade</td>
			<td>ThermoFisher Scientific</td>
			<td>50-949-135</td>
			<td></td>
		</tr>
		<tr>
			<td>Nichrome wire</td>
			<td></td>
			<td></td>
			<td>Any supplier will do</td>
		</tr>
		<tr>
			<td>Electrical wires</td>
			<td></td>
			<td></td>
			<td>Any supplier will do</td>
		</tr>
		<tr>
			<td>Model paint</td>
			<td>Tamiya USA</td>
			<td>Depends on colour</td>
			<td></td>
		</tr>
		<tr>
			<td>Repeating dispenser</td>
			<td>Hamilton company</td>
			<td>PB-600-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass syringe</td>
			<td>WPI</td>
			<td>NANOFIL</td>
			<td></td>
		</tr>
		<tr>
			<td>flourescence viewing system</td>
			<td>Nightsea</td>
			<td>SFR-GR</td>
			<td></td>
		</tr>
		<tr>
			<td>graticule</td>
			<td>ProSciTech</td>
			<td>S8014-24</td>
			<td></td>
		</tr>
		<tr>
			<td>microcapillary with holder</td>
			<td>Drummond</td>
			<td>1-000-0010</td>
			<td></td>
		</tr>
		<tr>
			<td>Liquid silicone</td>
			<td></td>
			<td></td>
			<td>Any supplier will do</td>
		</tr>
		<tr>
			<td>Thermocouple</td>
			<td>Digitech</td>
			<td>QM-1324</td>
			<td></td>
		</tr>
		<tr>
			<td>Micropipette</td>
			<td>Eppendorf</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

neuropharmacological manipulation of restrained and free-flying honey bees, apis mellifera

Honningbier demonstrere forbløffende læreevne og avansert sosial atferd og kommunikasjon. I tillegg er deres hjerne liten, lett å visualisere og for å studere. Derfor bier har lenge vært et yndet modell blant nevrobiologer og neuroethologists for å studere nevrale grunnlaget for sosial og naturlig atferd. Det er imidlertid viktig at de eksperimentelle teknikker som brukes til å studere bier ikke forstyrrer virkemåter som studeres. På grunn av dette, har det vært nødvendig å utvikle en rekke teknikker for farmakologisk manipulering av honningbier. I denne artikkelen viser vi metoder for behandling av strekkfaste eller frittflygende honningbier med et bredt spekter av farmakologiske midler. Disse inkluderer både noninvasive metoder som muntlig og aktuelle behandlinger, samt mer invasive metoder som tillater presis levering av legemidler i enten systemisk eller lokal mote. Til slutt diskuterer vi fordeler og ulemper ved hver metode og beskrivefelles hekk og hvordan du best kan overvinne dem. Vi konkluderer med en diskusjon om viktigheten av å tilpasse den eksperimentelle metoden til de biologiske spørsmål snarere enn omvendt.

Et utvalg av representative resultater for de ovenfor beskrevne metoder er vist, primært for å demonstrere at fremgangsmåten tillater farmakologiske midler for å nå hjernen og påvirke honningbie oppførsel. Spesifikke effekter på hjerneprosesser kan lett skaffes etter thorax injeksjon. Fordi farmakologiske midler injiseres gjennom toraks kan virke på flere mål...

Watch this Scientific Journal Video about Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera at JoVE.com

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera

Dette manuskriptet beskriver flere protokoller for å administrere farmakologiske midler til honningbier, inkludert enkle noninvasive metoder for frittflygende bier, samt mer invasive varianter som tillater presis lokalisert behandling av behersket bier.

Nevrofarmakologiske Manipulering av Behersket og frittflygende Honey Bees,<em&gt; Apis mellifera</em

Honey bees demonstrate astonishing learning abilities and advanced social behavior and communication. In addition, their brain is small, easy to visualize ...

Research

JoVE Journal

Neuroscience

Neuropharmacological Manipulation of Restrained and Free-flying Honey Bees, Apis mellifera

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera)

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees Apis mellifera

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the ...

Mechanical Manipulation of Neurons to Control Axonal Development

Cell manipulations and extension of neuronal axons can be accomplished with calibrated glass micro-fibers capable of measuring and applying forces in the ...

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Development of a precise olfactory circuit relies on accurate projection of olfactory sensory neuron (OSN) axons to their synaptic 
targets in the ...

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees (Apis mellifera L.)

Tactile Conditioning And Movement Analysis Of Antennal Sampling Strategies In Honey Bees Apis mellifera L.

Tactile Conditioning and Movement Analysis Of antennal Prøvetaking Strategies i Honey Bees (<em&gt; Apis mellifera</em&gt; L.)

Honey bees (Apis mellifera L.) are eusocial insects and well known for their complex division of labor and associative learning capability1, 2. The worker ...

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Genetic Manipulation of the Mouse Developing Hypothalamus through In utero Electroporation

Genetisk manipulering av musen Utvikling Hypothalamus gjennom<em&gt; I utero</em&gt; Electroporation

Genetic  modification of specific regions of the developing mammalian brain is a very  powerful experimental approach. However, generating novel mouse ...

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees Apis mellifera

RNAi-mediert Double Gene Knockdown og Gustatory Perception Måling i Honey Bees (<em&gt; Apis mellifera</em&gt;)

This video demonstrates novel techniques of RNA  interference (RNAi) which downregulate two genes simultaneously in honey bees  using double-stranded RNA ...

Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

Genetic Manipulation of Cerebellar Granule Neurons In Vitro and In Vivo to Study Neuronal Morphology and Migration

Genetisk manipulering av Lillehjernen Granule Neuroner<em&gt; In Vitro</em&gt; Og<em&gt; I Vivo</em&gt; Å studere nevrale morfologi og migrasjon

Developmental events in the brain including neuronal morphogenesis and migration are highly orchestrated processes. In vitro and in vivo analyses allow ...

Juxtacellular Monitoring and Localization of Single Neurons within Sub-cortical Brain Structures of Alert, Head-restrained Rats

Juxtacellular Overvåking og lokalisering av enkelt nevroner innenfor subkortikale hjernestrukturer for offentlig varsling, Hode-behersket Rats

There are a variety of techniques to monitor extracellular activity of single neuronal units. However, monitoring this activity from deep brain structures ...

Inducing Plasticity of Astrocytic Receptors by Manipulation of Neuronal Firing Rates

Inducing Plasticity av astrocytic Reseptorer av Manipulering av nevronal utløsning priser

Close to two decades of research has established that astrocytes in situ and in vivo express numerous G protein-coupled receptors (GPCRs) that can be ...

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees (Bombus terrestris)

A Novel Behavioral Assay to Investigate Gustatory Responses of Individual, Freely-moving Bumble Bees Bombus terrestris

A Novel Behavioral analyse for å undersøke Gustatory Responses av individuelle, Fritt bevegelige Bumble Bees (<em&gt; Bombus terrestris</em&gt;)

Generalist pollinators like the buff-tailed bumble bee, Bombus terrestris, encounter both nutrients and toxins in the floral nectar they collect from ...