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Neuroscience

Foraging Path-length Protocol for Drosophila melanogaster Larvae

Published: April 23rd, 2016

DOI:

10.3791/53980

1Department of Ecology and Evolutionary Biology, University of Toronto, 2Department of Cell and Systems Biology, University of Toronto, 3Child and Brain Development Program, Canadian Institute for Advanced Research

We provide a detailed protocol for a Drosophila melanogaster foraging path-length assay. We discuss the preparation and handling of test animals, how to perform the assay and analyze the data.

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral variation. The larval path-length assay was developed to measure individual differences in foraging behavior that were later linked to the foraging gene. Larval path-length is an easily scored trait that facilitates the collection of large sample sizes, at minimal cost, for genetic screens. Here we provide a detailed description of the current protocol for the larval path-length assay first used by Sokolowski. The protocol details how to reproducibly handle test animals, perform the behavioral assay and analyze the data. An example of how the assay can be used to measure behavioral plasticity in response to environmental change, by manipulating feeding environment prior to performing the assay, is also provided. Finally, appropriate test design as well as environmental factors that can modify larval path-length such as food quality, developmental age and day effects are discussed.

Since the discovery of the white gene in Thomas Hunt Morgan's laboratory in 1910, the fruit fly, Drosophila melanogaster (D. melanogaster), has been used as a model for the study of the molecular and physiological underpinnings of various biological processes. The popularity of D. melanogaster largely stems from the considerable quantity and variety of genetic tools. Drosophila's small size, relative ease of handling and short generation time render it an ideal model for genetics studies. Equally important is Drosophila's capacity to demonstrate many of the phenotypes expressed by more complex ....

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1. Prepare Grape Plates and Holding Bottles for Collection of Larvae

  1. To make holding bottles, cut holes in one side of 6 oz fly culture bottles, large enough to fit a fly vial plug for air supply (Fig. 1D).
  2. To make grape plates, prepare 250 ml of grape juice medium (1.8% agar, 45% grape juice, 2.5% acetic acid, 2.5% ethanol) by boiling the agar, grape juice and most of the water, cool down to 70 ºC (stir while cooling), then add acetic acid and ethanol and bring up to volu.......

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Differences in path-length of the rover and sitter for strains and the effect of food deprivation on path-length are illustrated in Fig. 3. Data collected over three consecutive days of testing showed a significant strain effect (F(1,421) = 351.89, p < 2.20 x 10-16; Fig. 3A), with rovers traveling farther than sitters. In addition to the strain effect, there was also a significant food treatment effect (F(1, 42.......

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The path-length assay outlined here offers a robust and simple measure of foraging behavior of Drosophila larvae. The protocol follows the general methodology described in Sokolowski2, but has since been improved in regards to efficiency and experimental controls. To the best of our knowledge this method is the only available method for measuring larval path-length. The original version of the path-length protocol2, 3, 15, 16 tested larvae on Petri dishes with a thin layer of yeast paste ap.......

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The authors acknowledge continued funding the Natural Sciences and Engineering Council of Canada (NSERC) to MBS.

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Name Company Catalog Number Comments
6 oz  fly culture bottles  Fisher Scientific  AS355 
Fly vial plugs Droso-Plugs 59-201
35X10mm Petri dishes  Falcon 351008
100X15 mm Petri dishes  Fisher 875712
60x15mm Petri dishes VWR 25384-168 
Dissecting probes Almedic 2325-58-5300 
Yeast Lab Scientific FLY-8040-20F

  1. Dubnau, J. . Behavioral Genetics of the Fly (Drosophila melanogaster). , 173 (2014).
  2. Sokolowski, M. B. Foraging strategies of Drosophila melanogaster: a chromosomal analysis. Behav Genet. 10, 291-302 (1980).
  3. de Belle, J. S., Hilliker, A. J., Sokolowski, M. B. Genetic localization of foraging (for): A major gene for larval behavior in Drosophila melanogaster. Genetics. 123, 157-164 (1989).
  4. Osborne, K. A., Robichon, A., Burgess, E., Butland, S., Shaw, R. A., Coulthard, A., Pereira, H. S., Greenspan, R. J., Sokolowski, M. B. Natural behavior polymorphism due to a cGMP-dependent protein kinase of Drosophila. Science. 277, 834-836 (1997).
  5. Kalderon, D., Rubin, G. cGMP-dependent protein kinase genes in Drosophila. J Biol Chem. 264 (18), 10739-10748 (1989).
  6. Reaume, C. J., Sokolowski, M. B. cGMP-dependent protein kinase as a modifier of behavior. Handb Exp Pharmacol. 191, 423-443 (2009).
  7. Schneider, C. A., Rasband, W. S., Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat Methods. 9, 671-675 (2012).
  8. Schindelin, J., et al. Fiji: an open-source platform for biological-image analysis. Nat Methods. 9 (7), 676-682 (2012).
  9. Pereira, H. S., MacDonald, D. E., Hilliker, A. J., Sokolowski, M. B. Chaser (Csr), a new gene affecting larval foraging behavior in Drosophila melanogaster. Genetics. 140, 263-270 (1995).
  10. Shaver, S. A., Riedl, C. A. L., Parkes, T. L., Sokolowski, M. B., Hilliker, A. J. Isolation of larval behavioral mutants in Drosophila melanogaster. J Neurogenet. 14, 193-205 (2000).
  11. Graf, S. A., Sokolowski, M. B. The rover/sitter Drosophila foraging polymorphism as a function of larval development, food patch quality and starvation. J Insect Behav. 2, 301-313 (1989).
  12. Gonzalez-Candelas, F., Mensua, J. L., Moya, A. Larval competition in Drosophila melanogaster: effects on development time. Genetics. 82, 33-44 (1990).
  13. Durisko, Z., Kemp, R., Mubasher, R., Dukas, R. Dynamics of social behavior in fruit fly larvae. PLoS One. 9 (4), e95495 (2014).
  14. Sawin, E. P., Harris, L. R., Campos, A. R., Sokolowski, M. B. Sensorimotor transformation from light reception to phototactic behavior in Drosophila larvae (Diptera: Drosophilidae). J Insect Behav. 7, 553-567 (1994).
  15. de Belle, J. S., Sokolowski, M. B. Heredity of rover/sitter: alternative foraging strategies of Drosophila melanogaster. Heredity. 59, 73-83 (1987).
  16. de Belle, J. S., Sokolowski, M. B., Hilliker, A. J. Genetic analysis of the foraging microregion of Drosophila melanogaster. Genome. 36, 94-101 (1993).
  17. Sokolowski, M. B., Pereira, H. S., Hughes, K. Evolution of foraging behavior in Drosophila by density dependent selection. PNAS. 94, 7373-7377 (1997).

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