Generating Drosophila Strains Expressing TRPA1

Flies were reared on standard cornmeal agar medium at room temperature (RT). The following fly strains were used: Canton-special, isogenic w¹¹¹⁸ (a gift from Marcus C. Stensmyr); elav-Gal4/CyO; trpA1¹ (BDSC#26504, backcrossed 5 times); 5xUAS-TRPA1-C (a gift from D. Tracey); tub-cyto-roGFP2-Orp1^{49 (link)}. To generate UAS-SmedTRPA1 flies, Smed-TRPA1 cDNA (see above) was cloned into pCR™8/GW/TOPO® TA (Invitrogen) and then transferred into a 40xUAS destination vector created introducing the Gateway® cassette into pJFRC8-40XUAS- IVS-mCD8::GFP (Addgene #26221) via the XhoI/ XbaI restriction sites. This construct was then used for embryo injection by BestGene Inc. to generate P[40XUAS::Smed-TRPA1]attP40 flies. Similarly, UAS-humanTRPA1 flies were obtained starting from a human TRPA1 cDNA (NP_015628; a gift from Mark Hoon) to generate P[40XUAS::hTRPA1]attP40 flies. Expression of the transgenes was confirmed by RT PCR. Full genotypes of fly stocks used in Figure 5: w¹¹¹⁸. w¹¹¹⁸; TRPA1¹. w¹¹¹⁸; elav-Gal4/+; TRPA1¹/TRPA1¹. w¹¹¹⁸; +/UAS-TRPA1-C; TRPA1¹/TRPA1¹. w¹¹¹⁸; elav-Gal4/UAS-TRPA1-C; TRPA1¹/TRPA1¹. w¹¹¹⁸; +/UAS-Smed-TRPA1; TRPA1¹/TRPA1¹. w¹¹¹⁸; elav-Gal4/UAS-Smed-TRPA1; TRPA1¹/TRPA1¹. w¹¹¹⁸; +/UAS-human TRPA1; TRPA1¹/TRPA1¹. w¹¹¹⁸; elav-Gal4/UAS-human TRPA1; TRPA1¹/TRPA1¹.

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Arenas O.M., Zaharieva E.E., Para A., Vásquez-Doorman C., Petersen C.P, & Gallio M. (2017). Activation of planarian TRPA1 by reactive oxygen species reveals a conserved mechanism for nociception. Nature neuroscience, 20(12), 1686-1693.

Publication 2017

pCR™8/GW/TOPO® TA pJFRC8-40XUAS- IVS-mCD8::GFP

Agar Cdna Embryo Genotypes Human Strains Topo Transgenes Vector

Corresponding Organization :

Other organizations : Northwestern University

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Variable analysis

independent variables

Fly strains used: Canton-special, isogenic w^1118, elav-Gal4/CyO; trpA1^1, 5xUAS-TRPA1-C, tub-cyto-roGFP2-Orp1^49, UAS-SmedTRPA1, UAS-humanTRPA1

dependent variables

Not explicitly mentioned

control variables

Flies were reared on standard cornmeal agar medium at room temperature (RT)

controls

Positive control: w^1118 (a gift from Marcus C. Stensmyr)
Negative control: w^1118; TRPA1^1

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