Aug 29, 2024

Public workspaceStartle induced negative geotaxis (SING)

DOI
dx.doi.org/10.17504/protocols.io.5jyl82637l2w/v1
  • 1VIB-KU Leuven Center for Brain & Disease Research, 3000 Leuven, Belgium.
Natalie Kaempf
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DOI: dx.doi.org/10.17504/protocols.io.5jyl82637l2w/v1
Protocol Citation: Natalie Kaempf, Uli Pech, Patrik Verstreken 2024. Startle induced negative geotaxis (SING). protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl82637l2w/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 29, 2024
Last Modified: August 29, 2024
Protocol Integer ID: 104252
Keywords: ASAPCRN, climbing assay, behavior, dopaminergic dysfunction, SING
Funders Acknowledgement:
Aligning Science Across Parkinson’s
Grant ID: ASAP-000430
EMBO long-term postdoctoral fellowship
Grant ID: ALTF_299-2019
Research project, FWO Vlaanderen
Grant ID: G0A5219N
Research project, FWO Vlaanderen
Grant ID: G0B8119N
Methusalem project
Grant ID: METH/21/05 (3M210778)
Research project, KU Leuven Parkinson Fonds
Grant ID: EQZ-PARFON-O2010
Opening the Future grant, Leuvens Universiteitsfonds (LUF)
Grant ID: EQZ-OPTFUP-O2010
Research project, FWO Vlaanderen
Grant ID: G031324N
Abstract
This protocol describes how to perform the Startle induced negative geotaxis (SING) assay. SING defects in parkinsonism fly models have previously been shown to be caused by dopaminergic impairments.
Startle induced negative geotaxis (SING)
Startle induced negative geotaxis (SING)
30s
30s
for the describtion of the SING apparatus please refer to:
Benzer, Seymour. 1967. “BEHAVIORAL MUTANTS OF Drosophila ISOLATED BY COUNTERCURRENT DISTRIBUTION.” Proceedings of the National Academy of Sciences 58 (3): 1112–19. https://doi.org/10.1073/pnas.58.3.1112.

Inagaki, Hidehiko K, Azusa Kamikouchi, and Kei Ito. 2010. “Methods for Quantifying Simple Gravity Sensing in Drosophila Melanogaster.” Nature Protocols 5 (1): 20–25. https://doi.org/10.1038/nprot.2009.196.
set a specific time period during the day (e.g. Zeitgeber 3-5) to assess negative geotaxis behavior, which will be kept the same for all following experiments
flies are transferred into the first tube of the apparatus without prior anesthesia and allowed to adjust for some minutes to experimental environment (24±1ºC, 40-60% humidity in behavior chamber, note down the environmental condition and time for every genotype)
tap the flies down five times and allowed to climb up for Duration00:00:30

30s
after these 30 s the flies that reached the upper tube are moved to the next tube
repeat the procedure 4 times
count the number N of (male/female) flies with the corresponding genotype per tube to calculate the SING score, which is normalized to the mean SING score of the control.

SING score = ((N1 *0) + (N2 *1)+ (N3 *2)+ (N4 *3)+ (N5 *4))/ 4(N1 + N2 + N3 + N4 + N5 )
Nk = number of flies in the kth tube
Protocol references
Benzer, Seymour. 1967. “BEHAVIORAL MUTANTS OF Drosophila ISOLATED BY COUNTERCURRENT DISTRIBUTION.” Proceedings of the National Academy of Sciences 58 (3): 1112–19. https://doi.org/10.1073/pnas.58.3.1112.

Inagaki, Hidehiko K, Azusa Kamikouchi, and Kei Ito. 2010. “Methods for Quantifying Simple Gravity Sensing in Drosophila Melanogaster.” Nature Protocols 5 (1): 20–25. https://doi.org/10.1038/nprot.2009.196.