Uas trpa1

Manufactured by Bloomington Drosophila Stock Center

Sourced in United States

The UAS-TrpA1 is a genetic tool used in Drosophila research. It allows for the temperature-dependent activation of neurons expressing the TrpA1 ion channel. This provides a means to manipulate neural activity in a controlled manner.

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10 protocols using uas trpa1

Drosophila neuronal and muscular reporters

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Following lines were used: OK371-Gal4 (genotype w¹¹¹⁸;OK371-Gal4;+, Bloomington #26160) for targeting glutamatergic neurons, MHC-tauGFP [22 (link)] as a muscle reporter line, MT11-Gal4 (genotype w;+;P{GawB}MT11)(Bloomington #37295), UAS-mCD8::mRFP (genotype y,w;UAS-mCD8::mRFP;+, Bloomington #27398 and y,w;+;UAS-mCD8::mRFP) (Bloomington #27399 (3^rd Chr.)),10xUAS-mCD8::GFP(genotype w;+;10xUAS-mCD8::GFP)(Bloomington #32184) UAS-TrpA1 (genotype w;UAS-TrpA1;+) (Bloomington #26263), UAS-shi^TS (w;+;UAS-shi^TS [39 (link)]), UAS-ChR2(H134R) (w;UAS-ChR2(H134R);+, Bloomington #28995), UAS-eNpHR (w;+;UAS-eNpHR::YFP, Bloomington #41752 gift from Leslie Griffith), UAS-PaGFP(A206K) (w;+;UAS-PaGFP(A206K), gift from A.S. Chiang [40 ]), UAS-GCamP3 (genotype w;+;UAS-GCamP3, Bloomington #32236), UAS-GCamP6s (genotype w¹¹¹⁸;UAS-GCamP6s;+, Bloomington #42746), OregonR (genotype +/w;+/UAS-TrpA1;+/+). For PaGFP experiments genotype was y,w/w;UAS-mCD8::mRFP; UAS-PaGFP(A206K).

Hückesfeld S., Schoofs A., Schlegel P., Miroschnikow A, & Pankratz M.J. (2015). Localization of Motor Neurons and Central Pattern Generators for Motor Patterns Underlying Feeding Behavior in Drosophila Larvae. PLoS ONE, 10(8), e0135011.

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Genetic Manipulation of Drosophila Neurobiology

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Flies were reared at 25°C on standard cornmeal-molasses medium, unless indicated otherwise. The following stocks were used in this study: GMR51F12-Gal4 (Jenett et al., 2012 (link)) (Bloomington Stock Center), GMR48A05-Gal4 (Jenett et al., 2012 (link)) (Bloomington Stock Center), Ilp7-Gal4 and HGN1-Gal4 were gifts from Dr Irene Miguel-Aliaga (Cognigni et al., 2011 (link)), UAS-ShiTS1 was a gift from Toshihiro Kitamoto (Kitamoto, 2013 (link)), UAS-TRPA1 (Bloomington Stock Center), 1XUAS-cd8::GFP ([Pfeiffer et al., 2010 (link)], Bloomington Stock Center), PPK1-Gal4 ([Ainsley et al., 2014 (link)], Bloomington Stock Center), UAS-PPK1 was a gift from Wayne Johnson (Ainsley et al., 2008 (link)), UAS-PPK1 RNAi #1 (Bloomington Stock Center), UAS-PPK1 RNAi #2 (Vienna Drosophila RNAi Center, 108683), UAS-PPK29 RNAi (Bloomington Stock Center), b88h49df (Bloomington Stock Center), A400df (Bloomington Stock Center) and yw flies.

Olds W.H, & Xu T. (2014). Regulation of food intake by mechanosensory ion channels in enteric neurons. eLife, 3, e04402.

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Drosophila Circadian Rhythm Mutants

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The following strains were used in this study: w¹¹¹⁸, Clk^Jrk, Pdf-Gal4, UAS-TrpA1, tubulin-Gal80^ts, Δ263b-Gal4/TM6B, Δ263a-Gal4/CyO, Bereft²⁴ [50 (link)], UAS-Bx and Bx loss-of-function mutation (Bx^hdpR26) [60 (link)]; UAS-mCD8::GFP, Bx-Gal4, Repo-Gal4, UAS-263b, and miR-263b^KO (obtained from the Bloomington Stock Center). All flies were raised on standard cornmeal/agar medium at 25 °C under 12-h:12-h LD cycle.

Nian X., Chen W., Bai W., Zhao Z, & Zhang Y. (2019). miR-263b Controls Circadian Behavior and the Structural Plasticity of Pacemaker Neurons by Regulating the LIM-Only Protein Beadex. Cells, 8(8), 923.

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Drosophila Neuronal Characterization Protocol

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Flies were cultured in standard cornmeal food at 25°C in 12h/12h light/dark cycles. Fly stocks include: w¹¹¹⁸, alrm-Gal4^{26 (link)}, alrm-LexA::GAD, UAS-wtrw-gfp, LexAop-R-GECO1 (Bloomington, 52224), tdc2-Gal4 (Bloomington, 9313), 20XUAS-IVS-GCaMP6s (Bloomington, 42746), 13XLexAop2-IVS-GCaMP6s-p10 (Bloomington, 44274), UAS-Kir2.1-EGFP (Bloomington, 6595), UAS-Oct-TyrR^RNAi (Bloomington, 28332), th-Gal4 (Bloomington, 8848), UAS-wtrw^VDRC (VDRC, 107423), Oct-TyrR^hono (Kyoto, 109038), UAS-shi^DN (Bloomington, 5822), UAS-TrpA1 (Bloomington, 26263), UAS-NpHR (Kyoto, 117008), UAS-CsChrimson (Bloomington, 55135), AdoR^KG03964ex (Bloomington, 30868), tdc2-LexA::p65^{27 (link)}, UAS-wtrw^2.23^{28 (link)}, wtrw^ex^{29 (link)}, tβh^nM18³⁰, tdc2^RO54³¹, th-LexA::p65³², tsh-Gal80³³.

Ma Z., Stork T., Bergles D.E, & Freeman M.R. (2016). Neuromodulators signal through astrocytes to alter neural circuit activity and behavior. Nature, 539(7629), 428-432.

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Fly Behavioral Assay Protocols

Cited 2 times

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Flies were raised at 25°C and 60% relative humidity on standard cornmeal fly food under 12:12h light:dark cycle. 4–7 days old flies were used for experiments and were transferred to fresh food vials 48h before behavioral tests. Fly population was randomized but was kept age matched in each trial. For food deprivation, flies were kept in empty bottles with a wet cotton plug to prevent desiccation. The following fly lines were ordered from Bloomington stock center: Npf-Gal4 (25681), npfr mutants (10747), 20XUAS-TTS-shi[ts1]-p10 (66600; referred to as UAS-shibire^ts1 in the text), UAS-TrpA1 (26263), MB461B (68327), MB370B (68319), MB504B (68329), MB296B (68308), MB320C (68253), MB077B (68283) and MB112C (68263). UAS-npf-RNAi (108772), UAS-npfr-RNAi (107663) and VT50658 (200166) were ordered from the Vienna Drosophila Resource Center. Other fly lines were described previously: redeye^{6 (link)}, R26E01^{15 (link)}, R35B12¹⁶, and UAS-CaLexA^{17 (link)}. The background control line was Canton-S (Heisenberg) strain.

Chouhan N.S., Griffith L.C., Haynes P, & Sehgal A. (2020). Availability of food determines the need for sleep in memory consolidation. Nature, 589(7843), 582-585.

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Fly Behavioral Assay Protocols

Cited 2 times

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Chouhan N.S., Griffith L.C., Haynes P, & Sehgal A. (2020). Availability of food determines the need for sleep in memory consolidation. Nature, 589(7843), 582-585.

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Genetic Tools for Drosophila Research

Cited 1 time

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If not stated otherwise, flies were kept at 25°C on either PF or YF under a 12hr light/12hr dark cycle. UAS:lrp1RNAi and UAS:lrp2RNAi lines are from Vienna Drosophila RNAi Center, and oregonRC, repo-Gal4, dilp2-Gal4, dilp2, UAS:gfpRNAi, UAS:GCaMP, UAS:trpA1 lines are from Bloomington Stock center. Published are UAS:ltpRNAi, ltp (Palm et al., 2012 (link)), imp-l2-Gal4 (Bader et al., 2013 (link)), lpp-Gal4 (Brankatschk and Eaton, 2010 (link)), lpr1, lpr2 (Khaliullina et al., 2009 (link)), lrp1, lrp2 (Riedel et al., 2011 (link)), moody-Gal4 (gift from C Klaembt), impl2-Gal4 (gift from E Hafen). UAS:cherry and rab3-Gal4 were generated by transforming VK37 or VK33 flies (Venken and Bellen, 2005 (link)) with our cloned constructs. Yrab3 and Yrab27 are N-terminally YFP^MYC tagged viable rab alleles (rab3 and rab27) generated by targeted YFP^MYC integration into the endogenous locus (publication in preparation).

Brankatschk M., Dunst S., Nemetschke L, & Eaton S. (2014). Delivery of circulating lipoproteins to specific neurons in the Drosophila brain regulates systemic insulin signaling. eLife, 3, e02862.

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Drosophila Genetic Lines Characterization

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The flies were kept at 25 °C unless otherwise stated. Line w¹¹¹⁸ was used as a wild-type line [21 (link)]. The following lines were obtained from the Bloomington Drosophila Stock Center: UAS-mCD8::GFP (#5137), UAS-NaChBac (#9468), and UAS-TrpA1 (#26264). The period-EGFP fly was generated in our previous study [22 (link)]. The pdf-Gal4 line was provided by Dr. Yong Zhang (University of Nevada, Reno, NV, USA).

Du X., Yu L., Ling S., Xie J, & Chen W. (2021). High-Salt Diet Impairs the Neurons Plasticity and the Neurotransmitters-Related Biological Processes. Nutrients, 13(11), 4123.

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Fly Lines for Genetic Manipulation

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The following fly lines were obtained from the Bloomington Drosophila Stock Center (Indiana University, Bloomington, IN, USA) and used in this study: Actin5c-Gal4 (BL25374), Tubulin84B-Gal4 (BL5138), elav-Gal4 (BL458), nSyb-Gal4 (BL51635), UAS-GFP-nls (nuclear localization signal; BL4775), UAS-Stinger (a super-bright GFP-nls variant; BL84277), UAS-mCD8::GFP (BL32184), UAS-myr::smGdP-V5 (BL62146), UAS-LacZ (BL1776), UAS-TrpA1 (BL26263), UAS-myr::GFP UAS-FRT-STOP-FRT-smHA (BL62128), UAS-FRT-STOP-FRT-TrpA1 (BL66871), and w[*]; P{w[+mC] = Ubi-p63E(FRT.STOP)Stinger}9F6 (BL32250). Flies were reared in bottles containing standard cornmeal agar and grown at 25°C with 70% relative humidity and a 12-hour light/12-hour dark cycle.

Merrill C.B., Titos I., Pabon M.A., Montgomery A.B., Rodan A.R, & Rothenfluh A. (2024). Iterative assay for transposase-accessible chromatin by sequencing to isolate functionally relevant neuronal subtypes. Science Advances, 10(13), eadi4393.

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Drosophila Neuronal Characterization Protocol

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Ma Z., Stork T., Bergles D.E, & Freeman M.R. (2016). Neuromodulators signal through astrocytes to alter neural circuit activity and behavior. Nature, 539(7629), 428-432.

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