Name: methods to assay drosophila behavior
Uploaded: 2012-03-07T13:00:00
Description: Watch this Scientific Journal Video about Methods to Assay Drosophila Behavior at JoVE.com

We would like to thank Astha Maltare for generating the larval crawling data. We would like to thank Dr. Nicholas Lanson Jr. for giving his comments on the manuscript. This work was supported by the Robert Packard Center for ALS at Johns Hopkins (to UBP) and the Amyotrophic Lateral Sclerosis Association (UBP); and R01MH083689 from the National Institutes of Mental Health (CDN).

1. Larval Crawling Assay
1. Larvae Collection
<ol>
	<li>Set up an 8 ounce bottle of flies (10-15 males + 10-15 females).</li>
	<li>Let flies lay eggs for 24 hours, then clear bottle of flies. (Transfer the adults into a new bottle and repeat as necessary).</li>
	<li>Incubate bottle for 3-4 days, or until third instar larvae are visible.</li>
	<li>Add 50 - 100 ml of 20% sucrose to the bottle with larvae and let sit for 20 minutes. Larvae will float to the top.</li>
	<li>Collect larvae using a 25...

<ol>
	<li>Pandey, U. B., Nichols, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+disease+models+in+Drosophila+melanogaster+and+the+role+of+the+fly+in+therapeutic+drug+discovery.">Human disease models in Drosophila melanogaster and the role of the fly in therapeutic drug discovery.</a> Pharmacol. Rev. 63 (2), 411-436 (2011).</li><li>Feany, M. B., Bender, W. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila+model+of+Parkinson's+disease.">A Drosophila model of Parkinson's disease.</a> Nature Mar. 23 (6776), 394-398 (2000).</li><li>Auluck, P. K., Bonini, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacological+prevention+of+Parkinson+disease+in+Drosophila.">Pharmacological prevention of Parkinson disease in Drosophila.</a> Nat. Med. 8 (11), 1185-1186 (2000).</li><li>Whitworth, A. J., Theodore, D. A., Greene, J. C., Benes, H., Wes, P. D., Pallanck, 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=Increased+glutathione+Stransferase+activity+rescues+dopaminergic+neuron+loss+in+a+Drosophila+model+of+Parkinson's+disease.">Increased glutathione Stransferase activity rescues dopaminergic neuron loss in a Drosophila model of Parkinson's disease.</a> Proc. Natl. Acad. Sci. U.S.A. 102 (22), 8024-8029 (2005).</li><li>Gargano, J. W., Martin, I., Bhandari, P., Grotewiel, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+iterative+negative+geotaxis+(RING):+a+new+method+for+assessing+age-related+locomotor+decline+in+Drosophila.">Rapid iterative negative geotaxis (RING): a new method for assessing age-related locomotor decline in Drosophila.</a> Exp. Gerontol. 40 (5), 386-395 (2005).</li><li>Lanson, N. A., Maltare, A., King, H., Smith, R., Kim, J. H., Taylor, J. P., Lloyd, T. E., Pandey, U. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila+model+of+FUS-related+neurodegeneration+reveals+genetic+interaction+between+FUS+and+TDP-43.">A Drosophila model of FUS-related neurodegeneration reveals genetic interaction between FUS and TDP-43.</a> Hum. Mol. Genet. 20 (13), 2510-2523 (2011).</li><li>Batlevi, Y., Martin, D. N., Pandey, U. B., Simon, C. R., Powers, C. M., Taylor, J. P., Baehrecke, E. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynein+light+chain+1+is+required+for+autophagy,+protein+clearance,+and+cell+death+in+Drosophila.">Dynein light chain 1 is required for autophagy, protein clearance, and cell death in Drosophila.</a> Proc. Natl. Acad. Sci. U.S.A. 107 (2), 742-747 (2010).</li><li>Sang, T. K., Chang, H. Y., Lawless, G. M., Ratnaparkhi, A., Mee, L., Ackerson, L. C., Maidment, N. T., Krantz, D. E., Jackson, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Drosophila+model+of+mutant+human+parkin-induced+toxicity+demonstrates+selective+loss+of+dopaminergic+neurons+and+dependence+on+cellular+dopamine.">A Drosophila model of mutant human parkin-induced toxicity demonstrates selective loss of dopaminergic neurons and dependence on cellular dopamine.</a> J. Neurosci. 27 (5), 981-992 (2007).</li><li>Stacey, S. M., Muraro, N. I., Peco, E., Labb&#233;, A., Thomas, G. B., Baines, R. A., van Meyel, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drosophila+glial+glutamate+transporter+Eaat1+is+regulated+by+fringe-mediated+notch+signaling+and+is+essential+for+larval+locomotion.">Drosophila glial glutamate transporter Eaat1 is regulated by fringe-mediated notch signaling and is essential for larval locomotion.</a> J. Neurosci. 30 (43), 14446-14457 (2010).</li><li>Repnikova, E., Koles, K., Nakamura, M., Pitts, J., Li, H., Ambavane, A., Zoran, M. J., Panin, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sialyltransferase+regulates+nervous+system+function+in+Drosophila.">Sialyltransferase regulates nervous system function in Drosophila.</a> J. Neurosci. 30 (18), 6466-6476 (2010).</li><li>Repnikova, E., Koles, K., Nakamura, M., Pitts, J., Li, H., Ambavane, A., Zoran, M. J., Panin, V. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sialyltransferase+regulates+nervous+system+function+in+Drosophila.">Sialyltransferase regulates nervous system function in Drosophila.</a> J. Neurosci. 30 (18), 6466-6476 (2010).</li><li>Nedelsky, N. B., Pennuto, M., Smith, R. B., Palazzolo, I., Moore, J., Nie, Z., Neale, G., Taylor, 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=Native+functions+of+the+androgen+receptor+are+essential+to+pathogenesis+in+a+Drosophila+model+of+spinobulbar+muscular+atrophy.">Native functions of the androgen receptor are essential to pathogenesis in a Drosophila model of spinobulbar muscular atrophy.</a> Neuron. 67 (6), 936-952 (2010).</li><li>Lorenzo, D. N., Li, M. G., Mische, S. E., Armbrust, K. R., Ranum, L. P., Hays, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectrin+mutations+that+cause+spinocerebellar+ataxia+type+5+impair+axonal+transport+and+induce+neurodegeneration+in+Drosophila.">Spectrin mutations that cause spinocerebellar ataxia type 5 impair axonal transport and induce neurodegeneration in Drosophila.</a> J. Cell Biol. 189 (1), 143-158 (2010).</li><li>Wang, J. W., Brent, J. R., Tomlinson, A., Shneider, N. A., McCabe, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ALS-associated+proteins+FUS+and+TDP-43+function+together+to+affect+Drosophila+locomotion+and+life+span.">The ALS-associated proteins FUS and TDP-43 function together to affect Drosophila locomotion and life span.</a> J. Clin. Invest. , (2011).</li><li>Choi, J. K., Jeon, Y. C., Lee, D. W., Oh, J. M., Lee, H. P., Jeong, B. H., Carp, R. I., Koh, Y. H., Kim, Y. 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+Drosophila+model+of+GSS+syndrome+suggests+defects+in+active+zones+are+responsible+for+pathogenesis+of+GSS+syndrome.">A Drosophila model of GSS syndrome suggests defects in active zones are responsible for pathogenesis of GSS syndrome.</a> Hum. Mol. Genet. 19 (22), 4474-4489 (2010).</li><li>Ruan, H., Wu, C. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Social+interaction-mediated+lifespan+extension+of+Drosophila+Cu/Zn+superoxide+dismutase+mutants.">Social interaction-mediated lifespan extension of Drosophila Cu/Zn superoxide dismutase mutants.</a> Proc. Natl. Acad. Sci. U.S.A. 105 (21), (2008).</li><li>Slawson, J. B., Kim, E. Z., Griffith, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+video+tracking+of+locomotion+in+adult+Drosophila+melanogaster.">High-resolution video tracking of locomotion in adult Drosophila melanogaster.</a> J. Vis. Exp. (24), (2009).</li><li>Becnel, J., Johnson, O., Luo, J., N&#228;ssel, D. R., Nichols, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+serotonin+5-HT7+Dro+receptor+is+expressed+in+the+brain+of+Drosophila,+and+is+essential+for+normal+courtship+and+mating.">The serotonin 5-HT7 Dro receptor is expressed in the brain of Drosophila, and is essential for normal courtship and mating.</a> PLoS One. 6 (6), e20800 (2011).</li><li>Johnson, O., Becnel, J., Nichols, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonin+5-HT(2)+and+5-HT(1A)-like+receptors+differentially+modulate+aggressive+behaviors+in+Drosophila+melanoga-+ster.">Serotonin 5-HT(2) and 5-HT(1A)-like receptors differentially modulate aggressive behaviors in Drosophila melanoga- ster.</a> Neuroscience. 158 (2), 1292-1300 (2009).</li><li>Bastock, M., Manning, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Courtship+of+Drosophila+Melanogaster.">The Courtship of Drosophila Melanogaster.</a> Behaviour. , 85-111 (1955).</li><li>Greenspan, R. J., Ferveur, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Courtship+in+Drosophila.">Courtship in Drosophila.</a> Annu. Rev. Genet. 34, 205-232 (2000).</li><li>Villella, A., Hall, 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=Neurogenetics+of+courtship+and+mating+in+Drosophila.">Neurogenetics of courtship and mating in Drosophila.</a> Adv. Genet. 62, 67-184 (2008).</li></ol>

Drosophila behavior is tightly regulated by genetic and environmental factors. We, and others, have previously used the assays described here to gather data to examine genes related to fly behaviors and to human neurodegenerative diseases modeled in Drosophila5-19. For the crawling assay, careful selection of 3rd instar larvae is a critical step. If treating with a drug, it will take 10-15 minutes (or more depending on the type and nature of the drug) to achieve maximal effect if i...

<table border="1">
 <tbody>
 <tr>
 <td>Name of the reagent</td>
 <td>Company</td>
 <td>Catalogue number</td>
 </tr>
 <tr>
 <td>Sucrose</td>
 <td>Fisher Scientific</td>
 <td>S5-500</td>
 </tr>
 <tr>
 <td>Agarose</td>
 <td>Invitrogen</td>
 <td>16500-500</td>
 </tr>
 <tr>
 <td>6 oz Drosophila bottle</td>
 <td>Genesee Scientific</td>
 <td>32-130</td>
 </tr>
 <tr>
 <td>Paint Brush (#1)</td>
 <td>Ted Pella,Inc.</td>
 <td>11859 </td>
 </tr>
 <tr>
 <td>Fly food components</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Cornmeal</td>
 <td>Fisher Scientific</td>
 <td>NC9109741</td>
 </tr>
 <tr>
 <td>Agar</td>
 <td>Genesee Scientific</td>
 <td>66-104</td>
 </tr>
 <tr>
 <td>Molasses</td>
 <td>Fisher Scientific</td>
 <td>NC9349176</td>
 </tr>
 <tr>
 <td>Propionic acid</td>
 <td>Acros</td>
 <td>14930-0010</td>
 </tr>
 <tr>
 <td>Tegosept</td>
 <td>Apex</td>
 <td>20-258</td>
 </tr>
 <tr>
 <td>Ethanol</td>
 <td>Fisher Scientific</td>
 <td>BP2818-4</td>
 </tr>
 <tr>
 <td>Yeast</td>
 <td>Genesee Scientific</td>
 <td>62-107</td>
 </tr>
 </tbody>
</table>

methods to assay drosophila behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Watch this Scientific Journal Video about Methods to Assay Drosophila Behavior at JoVE.com

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

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Neuroscience

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