Gustatory avoidance of fatty acids by Aedes aegypti depends on an arthropod-specific TRP channel
Data files
Jan 27, 2026 version files 2.28 GB
-
Raw_Data.zip
2.23 GB
-
README.md
9.25 KB
-
Source_Data.zip
48.19 MB
Abstract
Mosquito-disease vectors, such as Aedes aegypti, use their sense of taste before deciding whether to consume a blood meal, or fly away. However, the molecular mechanisms controlling gustatory decisions in mosquitoes are largely unknown. The TRP channel, Painless1 (Pain1), is an intriguing candidate for participating in Ae. aegypti taste since pain1 transcripts are detected in gustatory receptor neurons (GRNs). The Drosophila homolog, painless (pain) is also expressed in GRNs, where it is required for sensing allyl-isothiocyanate. Here, to identify additional gustatory roles for pain homologs, we first focused on Drosophila pain, which is widely expressed in multiple GRNs classes. We demonstrated that pain mutations eliminated gustatory attraction to low fatty acids levels, repulsion to high levels, and fatty acid-induced action potentials. The attraction and repulsion depended on pain expression in different GRN classes. In contrast to Drosophila, when Aedes contacts fatty acids, they induce gustatory rejection only. Aedes pain1 is expressed in taste organs, and is required for gustatory avoidance of fatty acids, and for fatty acid-induced action potentials. Given that Pain homologs are found in insects but not vertebrates, Pain1 represents an intriguing target for developing repellents to diminish biting, and transmission of infectious agents by mosquito disease vectors.
Dataset DOI: 10.5061/dryad.qrfj6q5ww
Description of the data and file structure
These data support the manuscript entitled “Gustatory avoidance of fatty acids by Aedes aegypti depends on an arthropod-specific TRP channel.” The dataset consists of two compressed folders, Raw_Data.zip and Source_Data.zip, which together contain the raw and processed data underlying the figures and supplementary figures presented in the manuscript.
The deposited data include the raw data corresponding specifically to the images, traces, and gels displayed in the manuscript, as well as the full quantitative source data used for statistical analyses and figure generation. Only raw data corresponding to the representative images, traces, and gels shown in the figures are deposited; raw data from additional experimental replicates generated during the study are not included.
Files and variables
File: Raw_Data.zip
Description: The Raw_Data.zip archive contains raw data files corresponding specifically to the representative images and traces displayed in the main and supplementary figures of the manuscript. This archive does not include all raw data generated during the study or all experimental replicates.
The contents include raw confocal microscopy image files corresponding to the images shown in the figures, representative electrophysiological tip-recording raw traces displayed in the manuscript, uncropped scanning electron microscopy (SEM) images corresponding to SEM images shown in the figures, and uncropped RT-PCR gel images corresponding to gel images displayed in the manuscript. Only raw files associated with the representative data shown in the figures are provided. Quantitative analyses and biological replicates derived from these and additional raw data are provided in the source data files.
Description of individual raw data files
Raw data_Fig. 1A–O
Raw confocal microscopy image files underlying Figure 1A–O, illustrating the anatomical organization, expression patterns, and overlap of gustatory receptor neurons in the labellum.
Raw data_Fig. 2K, L, M, N
Representative raw extracellular tip-recording traces underlying Figure 2K–N, showing neuronal responses of labellar taste sensilla to fatty acids and control tastants.
Raw data_Fig. 3B
Uncropped RT-PCR gel images underlying Figure 3B, corresponding to the gel image displayed in the manuscript for gene expression validation.
Raw data_Fig. 4A–D
Raw confocal microscopy image files underlying Figure 4A–D, illustrating neuronal expression patterns and experimental manipulations affecting gustatory signaling.
Raw data_Fig. 6A
Representative SEM image of mosquito labellum each for dorsal and ventral view respectively.
Raw data_Fig. 6E–G and 6I–K
Representative raw extracellular tip-recording traces underlying Figure 6E–G, illustrating changes in gustatory neuron activity across experimental conditions.
Raw data folders for supplementary figures
Raw data_Supplementary Fig. 1A–C
Raw confocal microscopy image files underlying Supplementary Figure 1A–C, providing additional anatomical and expression data supporting Figure 1.
Raw data_Supplementary Fig. 2D, E, G, H, J, K, M, N
Representative raw extracellular tip-recording traces underlying the indicated panels of Supplementary Figure 2.
Raw data_Supplementary Fig. 3A, B, D, E, G, H
Raw confocal microscopy image files underlying Supplementary Figure 3, providing additional neuronal labeling and expression analyses.
Raw data_Supplementary Fig. 4E–H
Uncropped RT-PCR gel images underlying Supplementary Figure 4E–H.
Raw data_Supplementary Fig. 7E
Representative raw extracellular tip-recording traces underlying Supplementary Figure 7E.
File: Source_Data.zip
Description: The Source_Data.zip archive contains all quantitative source data used to generate the figures in the manuscript, provided in Excel format. These data include values derived from all biological replicates used for statistical analysis, even when only representative raw images or traces are shown in the figures.
The archive contains two subfolders. The Excel_Sheets folder contains numerical source data used for plotting and statistical analyses. The Images folder contains uncropped but processed images used in Fig. 1A–O and Supplementary Fig. 1A–C. Each figure has a corresponding Excel file labeled by figure name. For example, the source data file for Figure 2A is named Source_Data_Fig_2A.xlsx, and the file for Supplementary Figure 3A is named Source_Data_Supplementary_Fig_3A.xlsx. All experimental details are described in the Methods and figure legend sections of the manuscript.
Description of individual source data files
Source data files labeled Source Data_Fig. 2A–I contain data obtained from proboscis extension response (PER) assays performed in flies exposed to different chemical stimuli. Each value represents the response of an individual fly, with “1” indicating a positive PER and “0” indicating a negative PER. Rows correspond to experimental variables, and columns correspond to groups of flies. Each column represents one biological replicate (N = 1), consisting of a group of flies.
The Source Data_Fig. 2J file contains data obtained from tip-recording assays performed on labellar taste hairs of flies exposed to the indicated concentrations of tastants. Each recording lasted 10 seconds, but only the first 1 second was analyzed. Rows indicate experimental variables, and columns represent individual flies used for recordings. Representative raw traces displayed in the manuscript are provided in the Raw_Data.zip archive.
The Source Data_Supplementary Fig. 4C file contains data obtained from quantitative real-time PCR (RT-qPCR) experiments. PCR was performed using a Bio-Rad qPCR system, and Ct values are reported for each gene. Ribosomal protein S7 (RPS7) was used as the internal control. Uncropped RT-PCR gel images corresponding to those displayed in the manuscript are provided in the Raw_Data.zip archive.
Source data files Source Data_Fig. 5B–C contain data obtained from a single blood-feeding assay using adult female mosquitoes. The values represent the percentage of mosquitoes engorged on blood feeders with or without chemical treatments. Rows indicate experimental variables, and columns represent biological replicates.
Source data files Source Data_Fig. 5F–I contain data obtained from two-way choice blood-feeding assays using adult female mosquitoes. These files include the number of mosquitoes choosing either a control blood feeder or a blood feeder coated with fatty acids or other chemicals. Preference Index (PI) values were calculated from these counts. Rows indicate experimental variables, and columns represent biological replicates.
Source data files Source Data_Fig. 6H and 6L contain data obtained from tip-recording assays performed on labellar taste hairs of female mosquitoes. Each recording lasted 10 seconds, but only the first 500 milliseconds were analyzed. Rows indicate experimental variables, and columns represent individual female mosquitoes used for recordings. Representative raw traces displayed in the manuscript are included in the Raw_Data.zip archive.
Source data files Source Data_ Supplementary Fig. 8
Source data underlying Supplementary Figure 8, containing quantified calcium imaging responses from HEK293 cells heterologously expressing Drosophila melanogaster Pain or Aedes aegypti Pain1 channels. The data represent normalized changes in intracellular calcium levels measured following stimulation, and were used to generate the plots shown in the supplementary figure. Rows represents cell used for the analysis.
The Source_Data_Table_1 file contains data generated from confocal microscopy analyses. Immunohistochemistry was performed on fly labella with counterstaining to visualize distinct gustatory receptor neurons (GRNs), and samples were imaged using a confocal microscope. Neurons were counted to assess overlap between different GRN populations. Representative confocal raw images displayed in the figures are included in the Raw_Data.zip archive. Rows indicate variables, and columns represent the number of samples analyzed.
Other source data Excel files for main and supplementary files follow similar formats and can be interpreted using the descriptions above.
Acronyms:
LA: Linoleic acid
HA: Hexanoic acid
BA: Butyric acid
DA: Decanoic acid
1 s: 1 second
500 ms: 500 milliseconds
PER: Proboscis extension response
TR: Tip recording
WT: Wild-type
PI: Preference Index
Ave: Average of PI
SEM: Standard error of mean
Code/software
Confocal microscopy data were acquired using a Zeiss LSM 900 confocal microscope. CZI files can be opened using ImageJ/Fiji or Zeiss ZEN software. Tip-recording data were acquired using AutoSpike software (version 3.1; Syntech; https://www.ockenfels-syntech.com/download-2-2/) and require this software for viewing and analysis.
Data were generated using established behavioral, electrophysiological, molecular, and imaging approaches in Drosophila melanogaster and Aedes aegypti, as described in detail in the associated article. Proboscis extension response (PER) assays were performed by stimulating the labella or tarsi with sucrose alone or sucrose mixed with fatty acids or other tastants, and scoring feeding responses. Electrophysiological recordings were obtained using extracellular tip-recording techniques from individual labellar taste sensilla following chemical stimulation, and action potential frequencies were quantified over defined analysis windows. Gene expression was assessed using RT-PCR and quantitative real-time PCR (RT-qPCR), with ribosomal protein genes used as internal controls. Confocal microscopy was used to image gustatory receptor neuron populations following immunohistochemistry or reporter expression, and scanning electron microscopy (SEM) was used to characterize labellar sensilla morphology. Full experimental protocols, sample sizes, and statistical methods are provided in the main article Methods and figure legends.