TY - JOUR
T1 - An improved assay and tools for measuring mechanical nociception in drosophila larvae
AU - Lopez-Bellido, Roger
AU - Galko, Michael J.
N1 - Funding Information:
We thank Thomas Wang for developing the prototype von Frey filaments, Patrick J. Huang for improving the mechanical probe assay, the Bloomington Drosophila Stock Center for the control (w1118) and ppk-Gal4>UAS-mCD8-GFP fly stocks, and Galko lab members for critically reading the manuscript. This work was supported by R21NS087360 and R35GM126929 to MJG.
Publisher Copyright:
© 2020, Journal of Visualized Experiments. All rights reserved.
PY - 2020/10
Y1 - 2020/10
N2 - Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.
AB - Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.
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U2 - 10.3791/61911
DO - 10.3791/61911
M3 - Article
C2 - 33191934
AN - SCOPUS:85096254185
SN - 1940-087X
VL - 2020
JO - Journal of Visualized Experiments
JF - Journal of Visualized Experiments
IS - 164
M1 - e61911
ER -