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 för utnyttjas bin <ol><li> oral behandling <ol><li> Förbereda 1,5 M sackaroslösning genom blandning av 257 g sackaros med 500 ml vatten (det är lättare att lösa upp denna mängd sackaros i kokande vatten). Lagra sackaroslösning vid 4 ° C fram till användning. OBS: Sucrose lösning ger en mycket gästfri miljö för vissa mikroorganismer, och därmed lätt blir förorenat och obehagliga för bin. Bulk sackaroslösning kan alikvoter och lagrades vid -20 ° C fram t...

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De metoder som beskrivs ovan tillåter enkelt, effektivt och robust behandling av antingen fritt flygande eller bör tas i bruk honungsbin. Dessa metoder är kompatibla med många experimentella paradigm och biologiska frågor (tabell 1). Alla de fritt flygande metoder kan lätt appliceras på bör tas i bruk bin. Det omvända är mindre framgångsrik, men eftersom tillfälliga återhållsamhet och invasiva behandlingsmetoder ofta kan äventyra binas flygande förmåga. Meto...

Eftersom Karl von Frisch klar sin dans språk 1, har honungsbin förblivit en populär studie art för forskare i djurens beteende och neurobiologi. Under de senaste åren en myriad av nya discipliner har dykt upp i skärningspunkten mellan dessa två områden, och flera andra discipliner (t.ex., molekylärbiologi, genomik och datavetenskap) har uppstått vid sidan av dem. Detta har lett till en snabb utveckling av nya teorier och modeller för att förstå hur beteende är resultatet av aktivitet i nervsystemet. På grund av den unika livsstil, rika beteenderepertoar och enkel experimentell och farmakologisk manipulation, har bin kvar i spetsen för denna revolution. Honungsbin används för att studera grundläggande neurobiologiska frågor som de underliggande inlärning och minne 2,3, beslutsfattande 4, lukt 5, eller visuell bearbetning sex. Under de senaste åren, HONey bee har även använts som en modell för att studera ämnen i allmänhet reserverade för medicinsk forskning, såsom effekterna av beroendeframkallande droger 7 - 11, sova 12, åldrande 13 eller de mekanismer som ligger till grund för anestesi 14. Till skillnad från de klassiska genetiska modellorganismer (t.ex. D. melanogaster, C. elegans, M. musculus), det finns mycket få genetiska verktyg för att manipulera neurala funktioner i honungsbin, även om detta är närvarande förändras 15. I stället har honungsbiet studier främst förlitat sig på farmakologiska manipulationer. Detta har varit mycket framgångsrik; dock, är den mångfald av bee forskning så att en rad metoder för farmakologisk administration behövs. Forskning med honungsbin behandlar vitt skilda frågor, studeras av forskare från olika discipliner och bakgrunder, och använder en mängd olika experimentella metoder. många research frågor kräver bin till antingen fritt flygande, fritt interagera i sin koloni, eller båda. Detta kan göra det svårt att hålla reda på enskilda försöksdjur, och gör återhållsamhet eller kanyle omöjligt. För att tillgodose de olika honungsbiet forskning behövs ett flertal drug delivery-metoder, vilket möjliggör robust och flexibel administration samtidigt som den farmakokinetiska och farmakodynamiska profilerna, invasivitet av metoden och dess tillförlitlighet, passa paradigm i fråga. På grund av dessa olika behov, har de flesta forskargrupper utvecklat sina egna unika läkemedelsadministreringsmetoder. Hittills har detta varit en styrka av biet forskarsamhället; Det har lett till utvecklingen av uppsättningar av metoder som gör det möjligt för administration av samma läkemedel i olika situationer. Vårt mål är inte att utveckla en enda standardiserad metod för farmakologiska manipulationer av bin, utan snarare att belysa metoder somhar visat sig vara särskilt framgångsrik, och hjälpa forskare antar dessa. Vi diskuterar de grundläggande principerna för hur de fungerar, samt deras fördelar och nackdelar.

<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

Honungsbin visar häpnadsväckande inlärningsförmåga och avancerad socialt beteende och kommunikation. Dessutom är deras hjärna liten, lätt att visualisera och att studera. Därför har bin länge varit en gynnad modell bland neurobiologists och neuroethologists för att studera neurala grunden för social och naturliga beteende. Det är dock viktigt att de experimentella tekniker som används för att studera bin inte stör de beteenden som studeras. På grund av detta har det varit nödvändigt att utveckla en rad tekniker för farmakologisk manipulation av honungsbin. I denna uppsats visar vi metoder för behandling av återhållen eller fritt flygande honungsbin med ett brett spektrum av farmakologiska medel. Dessa inkluderar både icke-invasiva metoder såsom orala och utvärtes behandlingar, liksom mer invasiva metoder som möjliggör exakt drug delivery antingen systemisk eller lokal mode. Slutligen, vi diskutera fördelar och nackdelar med varje metod och beskrivagemensamma hinder och hur man bäst övervinna dem. Vi avslutar med en diskussion om vikten av att anpassa den experimentella metoden till de biologiska frågor snarare än tvärtom.

Ett urval av representativa resultat för de metoder som beskrivits ovan visas, i första hand för att visa att metoderna tillåter farmakologiska medel för att nå hjärnan och påverka honungsbiet beteende. Specifika effekter på hjärnprocesser kan lätt erhållas efter bröstkorgen injektion. Eftersom farmakologiska medel injiceras genom bröstkorgen kan verka p?...

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

Detta manuskript beskriver flera protokoll för administrering av farmakologiska medel till honungsbin, inklusive enkla icke-invasiva metoder för fritt flygande bin, samt mer invasiva varianter som gör exakt lokaliserad behandling av återhållen bin.

Neurofarmakologiska Manipulering av återhållsamma och friflygande honungsbin,<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

Beteende farmakologi i klassiska Konditionering av SNABEL Extension Response på honungsbin ( 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

Mekanisk Manipulation av nervceller till Control axonal utveckling

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

Lentivirus-medierad genetisk manipulation och visualisering av Olfactory sensoriska neuroner In vivo</em

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.

Taktil konditionering och rörelse Analys av antenn provtagningsmetoder i honung Bin (<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

Genmanipulation av Mouse Utveckla Hypothalamus genom<em&gt; In utero</em&gt; Elektroporering

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-medierad Dubbel Gene Knockdown och Smaklig Perception Mätning honungsbin (<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

Genetic Manipulation av Cerebellar granulneuroner<em&gt; In Vitro</em&gt; Och<em&gt; In Vivo</em&gt; Att studera neuronal morfologi och migration

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 Övervakning och lokalisering av Single nervceller inom Sub-kortikala hjärnstrukturer av registrering, Head-hållsamma råttor

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

Induktion Plasticitet hos astrocytiska Receptorer genom manipulering av neuron 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 analys för att undersöka smak Svaren från individuella, fritt rörliga humlor (<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 ...