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. 2021 Apr 23;13(6):462–469. doi: 10.1007/s13238-021-00845-8

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Three ionotropic receptors mediate fly’s distaste to zinc. (A) Two-way choice taste assay for a group of 50–80 wildtype files (Canton. S) per assay. Preference index of files between 5 mmol/L sucrose alone and the same concentration of sucrose with different concentrations of zinc gluconate. One-way ANOVA followed by post hoc t test with Bonferronicorrection; n = 4. (B) Two-way choice assays for screening candidate IR family receptors for aversive taste to zinc gluconate. GFP-RNAi was used as a control. One-way ANOVA followed by post hoc t test with Bonferroni correction; n = 4–8. (C–E) Dose-dependent taste aversion of Ir76b, Ir25a and Ir56b mutants to indicated concentration of zinc gluconate. Two-way ANOVA followed by post hoc t test with Bonferroni correction; n = 6–11. (F) Expression of Ir76b-QF (GFP) and Ir56b-Gal4 (tdTomato) on the labellum. (Scale bars: 50 μm.) (G) Expression of Ir25a-LexA (GFP) and Ir56b-Gal4 (tdTomato) on the labellum. (Scale bars: 50 μm.) (H) PER to 50mmol/L zinc gluconate in 4% sucrose of Ir56b mutant flies and genetic rescue of Ir56b in zinc GRNs. One-way ANOVA followed by post hoc t test with Bonferroni correction; n = 7–16. (I and J) PER of files with genetic knockdown of Ir25a, r56b or Ir76b to 50 mmol/L zinc gluconate supplement of 4% sucrose in Ir56b neurons (I) and ppk23 neurons (J), respectively. One-way ANOVA followed by post hoc t test with Bonferroni correction; n = 7–15. (K) Tip recordings from the indicated sensilla in response to different concentration of zinc gluconate. Two-way ANOVA followed by post hoc t test with Bonferroni correction; n = 5–8. (L) The representative traces of spikes from the indicated sensilla in response to different concentration of zinc gluconate. (M–P) Comparing WT with Ir76b, Ir25a or Ir56b mutant diminished the zinc-triggered spikes to zinc salt. One-way ANOVA followed by post hoc t test with Bonferroni correction; n = 8–10. (Q) Representative traces of spikes from indicated genotype of flies to 75 mmol/L zinc gluconate. For all comparisons performed in this figure, *P < 0.05, **P < 0.01