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 spændte Bier <ol><li> Oral behandling <ol><li> Forbered 1,5 M saccharoseopløsning ved blanding 257 g sucrose med 500 ml vand (det er lettere at opløse denne mængde sucrose i kogende vand). saccharoseopløsning ved 4 ° C indtil brug opbevares. BEMÆRK: Saccharose løsning giver en meget gæstfri miljø for visse mikroorganismer, og dermed let bliver forurenet og ubehagelig for bier. Bulk saccharoseopløsning kan alikvoteret og opbevaret ved -20 ° C indtil anvende...

<ol>
	<li>Frisch, K. .. . v. o. n. . B. e. e. s. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Their Vision, Chemical Senses, and Language. , (1971).</li><li>Giurfa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+amazing+mini-brain:+lessons+from+a+honey+bee.">The amazing mini-brain: lessons from a honey bee.</a> Bee World. 84 (1), 5-18 (2003).</li><li>Giurfa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+and+neural+analysis+of+associative+learning+in+the+honeybee:+a+taste+from+the+magic+well.">Behavioral and neural analysis of associative learning in the honeybee: a taste from the magic well.</a> J. Comp. Physiol. 193 (8), 801-824 (2007).</li><li>Perry, C. J., Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Honey+bees+selectively+avoid+difficult+choices.">Honey bees selectively avoid difficult choices.</a> Proc. Natl. Acad. Sci. U. S. A. 110 (47), 19155-19159 (2013).</li><li>Giurfa, M., Sandoz, J. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invertebrate+learning+and+memory:+Fifty+years+of+olfactory+conditioning+of+the+proboscis+extension+response+in+honeybees.">Invertebrate learning and memory: Fifty years of olfactory conditioning of the proboscis extension response in honeybees.</a> Learn. Mem. 19 (2), 54-66 (2012).</li><li>Srinivasan, M. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Honey+bees+as+a+model+for+vision,+perception,+and+cognition.">Honey bees as a model for vision, perception, and cognition.</a> Annu. Rev. Entomol. 55, 267-284 (2010).</li><li>S&#248;vik, E., Cornish, J. L., Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cocaine+tolerance+in+honey+bees.">Cocaine tolerance in honey bees.</a> PLoS One. 8 (5), e64920 (2013).</li><li>S&#248;vik, E., Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invertebrate+models+in+addiction+research.">Invertebrate models in addiction research.</a> Brain. Behav. Evol. 82 (3), 153-165 (2013).</li><li>S&#248;vik, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Reward processing and responses to drugs of abuse in the honey bee, Apis mellifera. , (2013).</li><li>S&#248;vik, E., Even, N., Radford, C. W., Barron, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cocaine+affects+foraging+behaviour+and+biogenic+amine+modulated+behavioural+reflexes+in+honey+bees.">Cocaine affects foraging behaviour and biogenic amine modulated behavioural reflexes in honey bees.</a> Peer J. 2, e662 (2014).</li><li>Abramson, C. I., Stone, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+an+ethanol+model+using+social+insects+I:+behavior+studies+of+the+honey+bee+(Apis+mellifera+L.).">The development of an ethanol model using social insects I: behavior studies of the honey bee (Apis mellifera L.).</a> Alcohol. Clin. Exp. Res. 24, 1153-1166 (2000).</li><li>Sauer, S., Kinkelin, M., Herrmann, E., Kaiser, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dynamics+of+sleep-like+behaviour+in+honey+bees.">The dynamics of sleep-like behaviour in honey bees.</a> J. Comp. Physiol. A Neuroethol. Sensory, Neural, Behav. Physiol. 189 (8), 599-607 (2003).</li><li>M&#252;nch, D., Kreibich, C. D., Amdam, G. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aging+and+its+modulation+in+a+long-lived+worker+caste+of+the+honey+bee.">Aging and its modulation in a long-lived worker caste of the honey bee.</a> J. Exp. Biol. 216 (Pt 9), 1638-1649 (2013).</li><li>Cheeseman, J. F., Winnebeck, E. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=General+anesthesia+alters+time+perception+by+phase+shifting+the+circadian+clock.">General anesthesia alters time perception by phase shifting the circadian clock.</a> Proc. Natl. Acad. Sci. , (2012).</li><li>Schulte, C., Theilenberg, E., M&#252;ller-Borg, M., Gempe, T., Beye, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+efficient+integration+and+expression+of+piggyBac-derived+cassettes+in+the+honeybee+(Apis+mellifera).">Highly efficient integration and expression of piggyBac-derived cassettes in the honeybee (Apis mellifera).</a> Proc. Natl. Acad. Sci. U. S. A. 111 (24), 9003-9008 (2014).</li><li>Felsenberg, J., Gehring, K. B., Antemann, V., Eisenhardt, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=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).</a> J. Vis. Exp. (47), e2282 (2011).</li><li>Burger, H., Ayasse, M., D&#246;tterl, S., Kreissl, S., Galizia, C. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perception+of+floral+volatiles+involved+in+host-plant+finding+behaviour:+Comparison+of+a+bee+specialist+and+generalist.">Perception of floral volatiles involved in host-plant finding behaviour: Comparison of a bee specialist and generalist.</a> J. Comp. Physiol. A Neuroethol. Sensory, Neural, Behav. Physiol. 199 (9), 751-761 (2013).</li><li>Pan, K. C., Goodman, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ocellar+projections+within+the+central+nervous+system+of+the+worker+honey+bee,+Apis+mellifera.">Ocellar projections within the central nervous system of the worker honey bee, Apis mellifera.</a> Cell Tissue Res. 176 (4), 505-527 (1977).</li><li>Ito, K., Shinomiya, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+nomenclature+for+the+insect+brain.">A systematic nomenclature for the insect brain.</a> Neuron. 81, 755-765 (2014).</li><li>Bitterman, M. E., Menzel, R., Fietz, A., Sch&#228;fer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classical+conditioning+of+proboscis+extension+in+honeybees+(Apis+mellifera).">Classical conditioning of proboscis extension in honeybees (Apis mellifera).</a> J. Comp. Psychol. 97 (2), 107-119 (1983).</li><li>Barron, A. B., Robinson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+modulation+of+task+performance+by+octopamine+in+honey+bee+(Apis+mellifera)+division+of+labour.">Selective modulation of task performance by octopamine in honey bee (Apis mellifera) division of labour.</a> J. Comp. Physiol. A Neuroethol. Sens. Neural. Behav. Physiol. 191 (7), 659-668 (2005).</li><li>Schulz, D. J., Sullivan, J. P., Robinson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Juvenile+Hormone+and+Octopamine+in+the+Regulation+of+Division+of+Labor+in+Honey+Bee+Colonies.">Juvenile Hormone and Octopamine in the Regulation of Division of Labor in Honey Bee Colonies.</a> Horm. Behav. 42 (2), 222-231 (2002).</li><li>Schulz, D. J., Elekonich, M. M., Robinson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biogenic+amines+in+the+antennal+lobes+and+the+initiation+and+maintenance+of+foraging+behavior+in+honey+bees.">Biogenic amines in the antennal lobes and the initiation and maintenance of foraging behavior in honey bees.</a> J. Neurobiol. 54 (2), 406-416 (2003).</li><li>Barron, A. B., Vander Meer, R. K., Maleszka, J., Robinson, G. E., Maleszka, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparing+injection,+feeding+and+topical+application+methods+for+treatment+of+honeybees+with+octopamine.">Comparing injection, feeding and topical application methods for treatment of honeybees with octopamine.</a> J. Insect Physiol. 53 (2), 187-194 (2007).</li><li>McClung, C., Hirsh, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stereotypic+behavioral+responses+to+free-base+cocaine+and+the+development+of+behavioral+sensitization+in+Drosophila.">Stereotypic behavioral responses to free-base cocaine and the development of behavioral sensitization in Drosophila.</a> Curr. Biol. 8 (2), 109-112 (1998).</li><li>Martin, B. R., Lue, L. P., Boni, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pyrolysis+and+volatilization+of+cocaine.">Pyrolysis and volatilization of cocaine.</a> J. Anal. Toxicol. 13 (3), 158-162 (1989).</li><li>Lefer, D., Perisse, E., Hourcade, B., Sandoz, J. -. C., Devaud, J. -. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+waves+of+transcription+are+required+for+long-term+memory+in+the+honeybee.">Two waves of transcription are required for long-term memory in the honeybee.</a> Learn. Mem. 20 (1), 29-33 (2012).</li><li>Urlacher, E., Soustelle, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Honey+Bee+Allatostatins+Target+Galanin/Somatostatin-Like+Receptors+and+Modulate+Learning:+A+Conserved+Function?.">Honey Bee Allatostatins Target Galanin/Somatostatin-Like Receptors and Modulate Learning: A Conserved Function?.</a> PLoS One. 11 (1), e0146248 (2016).</li><li>Stollhoff, N., Menzel, R., Eisenhardt, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+recovery+from+extinction+depends+on+the+reconsolidation+of+the+acquisition+memory+in+an+appetitive+learning+paradigm+in+the+honeybee+(Apis+mellifera).">Spontaneous recovery from extinction depends on the reconsolidation of the acquisition memory in an appetitive learning paradigm in the honeybee (Apis mellifera).</a> J. Neurosci. 25 (18), 4485-4492 (2005).</li><li>Barron, A. B., Maleszka, R., Helliwell, P. G., Robinson, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+cocaine+on+honey+bee+dance+behaviour.">Effects of cocaine on honey bee dance behaviour.</a> J. Exp. Biol. 212 (2), 163-168 (2009).</li><li>Devaud, J. -. M., Papouin, T., Carcaud, J., Sandoz, J. -. C., Gr&#252;newald, B., Giurfa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+substrate+for+higher-order+learning+in+an+insect:+Mushroom+bodies+are+necessary+for+configural+discriminations.">Neural substrate for higher-order learning in an insect: Mushroom bodies are necessary for configural discriminations.</a> Proc. Natl. Acad. Sci. , 1-9 (2015).</li><li>Vergoz, V., Roussel, E., Sandoz, J. -. C., Giurfa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aversive+learning+in+honeybees+revealed+by+the+olfactory+conditioning+of+the+sting+extension+reflex.">Aversive learning in honeybees revealed by the olfactory conditioning of the sting extension reflex.</a> PLoS One. 2 (3), e288 (2007).</li><li>Henry, M., B&#233;guin, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+common+pesticide+decreases+foraging+success+and+survival+in+honey+bees.">A common pesticide decreases foraging success and survival in honey bees.</a> Science. 336 (6079), 348-350 (2012).</li><li>S&#248;vik, E., Perry, C. J., LaMora, A., Barron, A. B., Ben-Shahar, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Negative+impact+of+manganese+on+honeybee+foraging.">Negative impact of manganese on honeybee foraging.</a> Biol. Lett. 11 (3), 20140989 (2015).</li><li>Farooqui, T., Vaessin, H., Smith, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Octopamine+receptors+in+the+honeybee+(Apis+mellifera)+brain+and+their+disruption+by+RNA-mediated+interference.">Octopamine receptors in the honeybee (Apis mellifera) brain and their disruption by RNA-mediated interference.</a> J. Insect Physiol. 50 (8), 701-713 (2004).</li><li>Guo, X., Su, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recipe+for+a+Busy+Bee:+MicroRNAs+in+Honey+Bee+Caste+Determination.">Recipe for a Busy Bee: MicroRNAs in Honey Bee Caste Determination.</a> PLoS One. 8 (12), e81661 (2013).</li><li>Cristino, A. S., Barchuk, A. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroligin-associated+microRNA-932+targets+actin+and+regulates+memory+in+the+honeybee.">Neuroligin-associated microRNA-932 targets actin and regulates memory in the honeybee.</a> Nat. Commun. 5, 5529 (2014).</li><li>Vargaftig, B. B., Coignet, J. L., de Vos, C. J., Grijsen, H., Bonta, I. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mianserin+hydrochloride:+Peripheral+and+central+effects+in+relation+to+antagonism+against+5-hydroxytryptamine+and+tryptamine.">Mianserin hydrochloride: Peripheral and central effects in relation to antagonism against 5-hydroxytryptamine and tryptamine.</a> Eur. J. Pharmacol. 16 (3), 336-346 (1971).</li><li>Beggs, K. T., Tyndall, J. D. A., Mercer, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Honey+bee+dopamine+and+octopamine+receptors+linked+to+intracellular+calcium+signaling+have+a+close+phylogenetic+and+pharmacological+relationship.">Honey bee dopamine and octopamine receptors linked to intracellular calcium signaling have a close phylogenetic and pharmacological relationship.</a> PLoS One. 6 (11), (2011).</li><li>Matsumoto, Y., Menzel, R., Sandoz, J. -. C., Giurfa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Revisiting+olfactory+classical+conditioning+of+the+proboscis+extension+response+in+honey+bees:+a+step+toward+standardized+procedures.">Revisiting olfactory classical conditioning of the proboscis extension response in honey bees: a step toward standardized procedures.</a> J. Neurosci. Methods. 211 (1), 159-167 (2012).</li></ol>

Metoderne er skitseret ovenfor tillader enkel, effektiv og robust behandling af enten frit flyvende eller styrkes med honningbier. Disse metoder er kompatible med mange eksperimentelle paradigmer og biologiske spørgsmål (tabel 1). Alle de fritflyvende metoder kan nemt anvendes på styrkes med bier. Det omvendte er mindre vellykket, men da midlertidig tilbageholdenhed og invasive behandlingsmetoder ofte kan kompromittere biernes flyvende evne. Metoderne er blevet præsenter...

Da Karl von Frisch belyst deres dans sprog 1, har honningbier forblevet en populær undersøgelse art for forskere i dyrs adfærd og neurobiologi. I de senere år et utal af nye discipliner er opstået i skæringspunktet mellem disse to områder, og flere andre discipliner (f.eks, molekylær biologi, genomforskning og datalogi) er opstået sammen med dem. Dette har ført til en hurtig udvikling af nye teorier og modeller til at forstå, hvordan adfærd skyldes aktivitet inden nervesystem. På grund af den unikke livsstil, rige adfærdsmæssige repertoire, og let eksperimenterende og farmakologisk manipulation, har bierne været på forkant med denne revolution. Honningbier bliver brugt til at studere grundlæggende neurobiologiske spørgsmål som dem, der ligger til grund for indlæring og hukommelse 2,3, beslutningstagning 4, olfaktoriske 5 eller visuel bearbejdning 6. I de seneste år har honey bi er endda blevet brugt som model til at studere emner generelt forbeholdt medicinsk forskning, såsom virkningerne af vanedannende stoffer 7 - 11, sove 12, ældning 13, eller de mekanismer, der ligger til grund anæstesi 14. I modsætning til de klassiske genetiske modelorganismer (f.eks D. melanogaster, C. elegans, M. musculus), der er meget få genetiske værktøjer til rådighed for at manipulere neurale funktioner i honningbier, selv om dette er i øjeblikket ændrer 15. I stedet har honningbi undersøgelser primært påberåbt farmakologiske manipulationer. Det har været meget vellykket; imidlertid, mangfoldigheden af ​​bi forskning er således, at der er brug for en række fremgangsmåder til farmakologisk administration. Forskning med honningbier adresser meget forskellige spørgsmål, er undersøgt af forskere fra forskellige discipliner og baggrunde, og bruger en række eksperimentelle tilgange. Mange research spørgsmål kræver bier til enten være fri-flyvende, frit interagere i deres koloni, eller begge dele. Dette kan gøre det svært at holde styr på de enkelte forsøgsdyr, og gør tilbageholdenhed eller kanylering umuligt. For at imødekomme de mange forskellige honningbi forskning, er der behov for en bred vifte af narkotika leveringsmetoder, der giver mulighed for robust og fleksibel administration og samtidig sikre, at den farmakokinetiske og farmakodynamiske profiler, invasionsevne af metoden, og dens pålidelighed, passer det paradigme pågældende. På grund af disse forskellige behov, har de fleste forskergrupper udviklet deres egne unikke Drug Administration metoder. Hidtil har dette været en styrke bi forskerkredse; det har ført til udviklingen af ​​arrays af fremgangsmåder, der muliggør administration af samme lægemiddel i forskellige omstændigheder. Vores mål her er ikke at udvikle en enkelt standardiseret metode til farmakologiske manipulationer af bier, men snarere at fremhæve metoder,har vist sig at være særdeles vellykket, og hjælpe forskere vedtage disse. Vi diskuterer de grundlæggende principper for, hvordan de arbejder, samt deres fordele 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 indlæringsevne og avanceret social adfærd og kommunikation. Desuden deres hjerne er lille, let at visualisere og studere. Derfor har bier længe været et yndet model blandt neurobiologer og neuroethologists til at studere det neurale grundlag for social og naturlige adfærd. Det er imidlertid vigtigt, at de eksperimentelle teknikker, der anvendes til at studere bier ikke forstyrrer den adfærd, der undersøges. På grund af dette, har det været nødvendigt at udvikle en række teknikker til farmakologisk manipulation af honningbier. I dette papir demonstrerer vi fremgangsmåder til behandling tilbage eller fritflyvende honningbier med en lang række farmakologiske midler. Disse omfatter både ikke-invasive fremgangsmåder, såsom orale og topiske behandlinger, såvel som mere invasive metoder, der giver mulighed for præcis lægemiddelafgivelse i enten systemisk eller lokaliseret måde. Endelig vil vi diskutere fordele og ulemper ved hver metode og beskrivefælles forhindringer og hvordan man bedst overvinde dem. Vi konkluderer med en diskussion om betydningen af ​​at tilpasse den eksperimentelle metode til de biologiske spørgsmål snarere end den anden vej rundt.

Et udvalg af repræsentative resultater for de ovenfor beskrevne metoder er vist, primært for at vise, at de metoder, tillader farmakologiske agenter til at nå hjernen og påvirke honningbi adfærd. Specifikke virkninger på hjernens processer kan let opnås efter thorax injektion. Fordi farmakologiske midler injiceres gennem brystkassen kan virke på flere mål i kr...

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 håndskrift beskriver flere protokoller til administration farmakologiske agenter til honningbier, herunder simple noninvasive metoder til fritflyvende bier, samt mere invasive varianter, der tillader præcis lokaliseret behandling af fastholdte bier.

Neurofarmakologisk Manipulation af Afdæmpet og fritflyvende honningbier<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 ...

The overall goal of these procedures is to reliable and robustly administer pharmacological agents to the central nervous system of honeybees or similar insects. These methods can help answer key questions in the fields of sociobiology, neuroethology, and apiculture such as exploring the neuromechanisms of social behavior. The main advantage of these techniques is that it makes it possible to study neurobiology in non-model organisms, such as the honeybee, where genetic tools are limited currently.Demontrating the procedure will be Jenny Plath, a graduate student from the Neuroethology Lab at Macquarie University. Drugs can be administered orally or by injection. Injections can be made into the thorax, into the ocellus, or into the ocellar tract.Ocellus injections deliver drugs to the whole head and ocellar tract injections target specific regions of the brain. For all injections, make a microscalpel by breaking off the corner of a double-edged razor blade with a blade holder. Attach the blade fragment to a blade holder so that it makes a nice blade with a sharp endpoint.For a thoracic injection dissect a two millimeter hole just above the scutellum, next to the posterior wing process of a bee's thorax. Avoid cutting too deep, as this might injure the flight muscles, and be careful to avoid the wing hinges. Ideally, only cut three sides so that the flap of the cuticle can later be folded back to close the site of the injury.Now inject the drug into the thorax. Then move the cuticle flap back into over the hole and after ten hours it will reattach and seal. To access the brain, lock the head of a harnessed bee in place by filling the neck crevice with hand-warmed dental wax in order to avoid damaging antennal olfactory receptors or other cells that may be important to the bee's behavior.The antenna may also be locked with dental wax. Then using the microscalpel, carefully remove the lens of the median ocellus by inserting the tip of the microscalpel under the lens and gently breaking the lens free from the head capsule. For the drug to be taken into the whole head capsule carefully pipette the drug onto the ocellus hole and wait for it to soak in.Alternatively, subregions of the brain can be targeted by injection through the ocellar tract, for example, by injecting 250 nanoliters of drug in a depth of 50 microns to target brain structures adjacent to the ocellus. Then remove the dental wax and allow the bee to rest for a while before continuing the experimental procedure. To prepare injection needles pull one millimeter glass tubes to produce an approximately 0.5 centimeter long tip.Next, under a stereo microscope break the tips to make and approximately 10 to 15 micron outer diameter opening. This can be measured using a scale on a graticule. Now load an injection needle with the drug solution.Then at the injection system, attach the loaded injection needle to the micromanipulator. Test the injection system in mineral oil and calibrate it to deliver a 0.5 to two nanoliter bolus depending on the size of the brain structure targeted. Using a graticule, measure the diameter of the droplets to calculate their volume.To fix the head of a harnessed bee use soft dental wax, as before. Then cut an opening into the anterior part of the head capsule using a microscalpel with three cuts:one just below the median ocellus;one at the border of the right or left eye;and one above the antenna stems. Use a piece of dental wax to hold the opened flap in place.Next carefully push aside the glands and trachea using fine forceps and make a small rupture into the neurilemma above the targeted brain structure. Now insert the tip of the glass pipette into the desired brain region, ideally, perpendicular to the brain surface into the required depth to reach the targeted structure. Then inject the drug bolus.When targeting paired brain regions make one injection into each hemisphere to affect both hemispheres. If a fluorescent dye is injected with the bolus the superficial brain regions can be viewed using fluorescent light. Other regions can be checked later using confocal microscopy.After the injection replace the flap of the cuticle and seal the cut by melting a crystal of eicosane that is approximately one millimeter in diameter over the cuticle using a thin wire attached to a microsoldering iron. This will greatly reduce mortality. To administer a drug as volatile as from a heated nichrome filament, prepare it dissolved in pure ethanol.To carefully drill holes in the lid of a 50 millimeter tube feed the two wires through the holes. To create a filament, wind up nichrome wire tightly around a nail, attach the nichrome wire to two electrical wires, and then remove the nail. Afterwards, secure the wires and seal the holes with silicone.Now attach the wires to a power source. And use a thermocouple to measure the temperature of the filament. To calibrate the device, carefully pipette two microliters of drug solution onto the filament and let the ethanol completely evaporate at room temperature.Vary the voltage in current so the drug bolus volatilizes within ten seconds of applying power. The drug should not be destroyed by thermal decomposition. Precipitate can usually be seen on the dry filament if inspected carefully.To deliver the drugs to a bee, load a free-flying or harnessed bee into the tube and carefully attach the lid with the wires out. Then turn on the power source for 10 seconds and wait 50 more seconds for the volatilized compound to be fully taken up by the bee before removing the bee. Specific effects on brain processes can be easily obtained following thorax injection.For example, blockers of transcription were administered using this technique to identify phases of memory that required gene expression. Selective and time-dependent affects on memory tasks were obtained. Diffusion of molecules into the head hemolymph leads to quick, dose-dependent effects arrived at by ocellus injection.At certain dose concentrations, diffusely-concentrated allatostatins reduced performance in an olfactory learning assay. To test the spatial and temporal properties of specific brain regions, harnessed bees were trained in an olfactory PER conditioning paradigm and then injected with procane in the mushroom body calices, or, vertical lobes. When bees were then tested for recall, performance was only impaired in bees injected bilaterally.In an arena with multiple feeders, bees were gently captured as they were about to alight from a feeder with one molar sucrose. The captured bees were treated with 100 micrograms of volatilized freebase cocaine, which resulted in increased foraging at half molar feeders, but not two molar feeders. After watching this video, you should have a good understanding of how to administer drugs to free-flying or restrained bees either by injection or volatilization.Once mastered, these techniques can easily be done on a large number of bees in a couple of hours. Following this procedure, other methods, like proboscis extension conditioning, can be performed so we can answer additional questions about the neurobiology of learning and memory.

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.

Taktil Conditioning og bevægelse Analyse af Antennal Sampling Strategies In 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 Manipulation af musen Udvikling Hypothalamus gennem<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-medieret Double Gene Knockdown og gustatoriske 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 manipulation af Cerebellær granuleneuroner<em&gt; In Vitro</em&gt; Og<em&gt; In Vivo</em&gt; At studere Neuronal morfologi og 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 Overvågning og lokalisering af Single neuroner inden subcorticale hjernestrukturer af indberetning, Head-behersket rotter

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

Inducerende plasticitet astrocytisk receptorer ved manipulation af neuronfyring 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 Assay til Undersøge gustatoriske Responses af Individuel, Frit bevægelse humlebier (<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 ...