Dataset DOI: 10.5061/dryad.cjsxksnnw

Description of the data and file structure

This dataset accompanies the manuscript "Gustatory function of Aedes aegypti opsins and TRPA1 in avoiding a natural insect repellant." It is organized into two compressed folders, Raw_data.zip and Source_data.zip, which together provide the raw and processed data underlying the figures and supplementary figures in the manuscript.

Raw_data.zip contains the original images, traces, and gels corresponding to the representative examples shown in the figures. Source_data.zip contains the full quantitative data used to generate the figures and perform the associated statistical analyses. Note that only the raw data underlying the specific representative images, traces, and gels displayed in the figures are included; raw data from additional experimental replicates collected during the study are not deposited here.

Files and variables

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 from all biological replicates used for statistical analysis, even in cases where only representative raw images or traces are shown in the figures.

The archive is organized into a single folder named "Source Data," which contains subfolders for each figure. Each subfolder includes Excel files with the numerical source data used for plotting and statistical analysis. Files are named according to their corresponding figure—for example, the source data for Figure 2A is named Fig. 2A.xlsx, and the source data for Supplementary Figure 3A is named Fig. S3A.xlsx.

Full experimental details for each dataset are described in the Methods and figure legend sections of the manuscript.

Description of individual source data files

Source data files Source Data Fig. 1A-D file contains data obtained from quantitative real-time PCR (RT-qPCR) experiments using proboscises, forelegs, midlegs and hindlegs. PCR was performed using a Bio-Rad qPCR system, and Ct values are reported for each gene. Ribosomal protein L17 (RPL17) was used as the internal control.

Source data files Source Data Fig. 2E-G file contains data obtained from the PRESTO-Tango assay. Catechin was tested at different concentrations in Op1, OP2 and OP1Op2 expressed HTLA cells, and the resulting change in luminescence was recorded, normalized, and reported as fold change.

Source data files Source Data Fig. 3A, C contains 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 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.3B contains 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 biological replicates, and columns represent experimental variables.

Source data files Source Data Fig. 3D contains 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 catechin and blood fed female mosquitoes were then evaluated for their aging over days. Rows indicate experimental variables, and columns represent biological replicates.

Source data files Source Data Fig. 3E contains data obtained from two-way choice egg laying assays using adult female mosquitoes. These files include the number of mosquitoes choosing either a control egg collection cup or egg collection cup containing catechin. Preference Index (PI) values were calculated from these counts. Rows indicate experimental variables, and columns represent biological replicates.

Source data files Source Data Fig. 3G contains data obtained from Y-tube olfactometer assays using adult female mosquitoes. These files include the number of mosquitoes choosing either a control side or side with catechin. Preference Index (PI) values were calculated from these counts. Rows indicate experimental variables, and columns represent biological replicates.

Source data files Source Data Fig.4S contains 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 file Source Data Fig. 5A-B contains data from tip-recording assays performed on labellar taste hairs of female mosquitoes. Each recording lasted 10 seconds, though only the first 500 milliseconds were analyzed. Rows represent replicates, and columns correspond to experimental variables.

Source data files Source Data Fig. 5G and 5I, contains calcium imaging responses from tarsi and labella of female mosquitoes. The data represent normalized changes in intracellular calcium levels measured following stimulation, and were used to generate the plots.

Source data files Source Data Fig. 5H and 5J, shows peak change in calcium responses from tarsi and labella of different female mosquitoes. The data represent normalized changes in intracellular calcium levels measured following stimulation. Rows represent replicates, and columns correspond to experimental variables.

Source data files Source Data Fig. 6F, shows peak change in current on applying different stimuli in two-electrode voltage clamp recordings expressing TRPA1 from different species. The data represent normalized changes in intracellular current levels measured following stimulation. Rows represent replicates, and columns correspond to experimental variables.

Source data files Source Data Fig. 6G contain data obtained from a single blood-feeding assay using adult female Anopheles gambiae mosquitoes. The values represent the percentage of mosquitoes engorged on blood feeders with or without chemical treatments. Rows indicate biological replicates, and columns represent experimental variables.

Source data files Source Data Fig. 7A-B contain data obtained from two-way choice sucrose feeding assays using adult fruit flies. These files include the number flies choosing either a control (sucrose) or sucrose plus chemical molecule. Preference Index (PI) values were calculated from these counts. Rows indicate biological replicates, and columns represent experimental variables.

Source data files labeled Source Data Fig. 7C-G contains 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.

Source data files Source Data Fig. 7H contains data from repellency assays testing the chemical at different concentrations against Ixodes scapularis on the human forearm. Rows represent replicates, and columns correspond to experimental variables.

Source data files Source Data Fig. 7K-L, shows peak change in current on applying different stimuli in two-electrode voltage clamp recordings expressing TRPA1 from different species. The data represent normalized changes in intracellular current levels measured following stimulation. Rows represent replicates, and columns correspond to experimental variables.

Source data files Source Data Fig. S1A-B file contains data obtained from quantitative real-time PCR (RT-qPCR) experiments using proboscises, and forelegs. PCR was performed using a Bio-Rad qPCR system, and Ct values are reported for each gene. Ribosomal protein L17 (RPL17) was used as the internal control.

Source data files Source Data Fig. S2A-B file contains data obtained from the PRESTO-Tango assay. Epicatechin and egcg was tested at different concentrations in Op1, OP2 expressed cells, and the resulting change in luminescence was recorded, normalized, and reported as fold change.

Source data files Source Data Fig. S3A and S3G contains 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 biological replicates, and columns represent experimental variables.

Source data files Source Data Fig. S3B shows data of porosity (phi %) comparison between two different types of membrane.

Source data files Source Data Fig. S3B shows data of elastic modulus (Er) comparison between two different types of membrane using microindentation.

Source data files Source Data Fig. S4A shows data from fourier transform infrared (FTIR) spectra of Vectaderm membrane and its individual constituents.

Source data files Source Data Fig. S4B shows data from thermogravimetric analysis (TGA) of Vectaderm membrane and its individual constituents.

Source data files Source Data Fig. S5A-C contains 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 biological replicates, and columns represent experimental variables.

Source data files Source Data Fig. S5D contains data obtained from two-way choice sucrose feeding assays using adult female mosquitoes. These files include the number of mosquitoes choosing either a control (sucrose) or sucrose with chemicals. Preference Index (PI) values were calculated from these counts. Rows indicate experimental variables, and columns represent biological replicates.

Source data file Source Data Fig. S6B-C, 6D and 6I contains data from tip-recording assays performed on labellar taste hairs of female mosquitoes. Each recording lasted 10 seconds, though only the first 500 milliseconds were analyzed. Rows represent replicates, and columns correspond to experimental variables.

Source data file Source Data Fig. S7D-E and 7N contains data from tip-recording assays performed on labellar taste hairs of fruit flies. Each recording lasted 10 seconds, though only the first 500 milliseconds were analyzed. Rows represent replicates, and columns correspond to experimental variables.

File: raw_data.zip

Description: The Raw_data.zip archive contains raw files corresponding specifically to the representative images and traces shown in the main and supplementary figures of the manuscript. It does not include all raw data generated during the study or all experimental replicates.

Its contents include raw confocal microscopy images corresponding to those shown in the figures; representative raw traces from electrophysiological tip-recording and TEVC experiments displayed in the manuscript; uncropped scanning electron microscopy (SEM) images; and uncropped RT-PCR gel images corresponding to the gel images displayed in the manuscript.

Raw data Fig. 1E–L
Raw confocal microscopy image files underlying Figure 1E-L, illustrating the anatomical organization, and expression patterns of neurons in the labellum.

Raw data Fig. 2C–D
Raw confocal microscopy image files underlying Figure 2C–D, illustrating the expression patterns of Op1 and Op2 in HEK-293 cells.

Raw data Fig. 4B
Uncropped RT-PCR gel images underlying Figure 4B, corresponding to the gel image displayed in the manuscript for gene expression validation.

Raw data Fig. 4C–R
Raw confocal microscopy image files underlying Figure 4C-R, illustrating the anatomical organization, and expression patterns of neurons in the labellum and tarsi.

Raw data Fig. 5C-F
Representative raw extracellular tip-recording traces underlying Figure 5C-F, showing neuronal responses of labellar taste sensilla to tastants.

Raw data Fig. 6B
Uncropped RT-PCR gel images underlying Figure 6B, corresponding to the gel image displayed in the manuscript for gene expression validation.

Raw data Fig. 6C-E
Raw two-electrode voltage clamp (TEVC) file underlying Figure 6C-E, corresponding to the image displayed in the manuscript for electrophysiology TEVC recordings.

Raw data Fig. 7I and 7J
Raw two-electrode voltage clamp (TEVC) file underlying Figure 7I and 7J, corresponding to the image displayed in the manuscript for electrophysiology TEVC recordings.

Raw data Fig. S1C-H
Raw confocal microscopy image files underlying Figure S1C-H, illustrating the anatomical organization, and expression patterns of neurons in the labellum.

Raw data Fig. S3C-D
Raw scanning electron microscopy (SEM) image files underlying Figure S3C-D, illustrating the porous structure in Vectaderm membrane and Hemotek membrane.

Raw data Fig. S3E
Raw image files underlying Figure S3E, illustrating the pH difference between Vectaderm membrane and commercial membrane.

Raw data Fig. S6A
Raw scanning electron microscopy (SEM) image files underlying Figure S6A, illustrating the sensilla arrangement on the labellum.

Raw data Fig. S6E-H and S6J-M
Representative raw extracellular tip-recording traces underlying Figure S6E-H and S6J-M, showing neuronal responses of labellar taste sensilla to tastants.

Raw data Fig. S7A, O and P
Uncropped RT-PCR gel images underlying Figure S7A, O and P, corresponding to the gel image displayed in the manuscript for gene expression validation.

Raw data Fig. S7B-C
Raw confocal microscopy image files underlying Figure S7B-C, illustrating the anatomical organization, and expression patterns of neurons in the labellum.

Raw data Fig. S7F-I and S7K-M
Representative raw extracellular tip-recording traces underlying Figure S7F-I and S7K-M, showing neuronal responses of labellar taste sensilla to tastants.

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. For TEVC recording open-source software WinWcp version 5.2.7 (https://spider.science.strath.ac.uk/sipbs/media/40/ses/WinWCP_V5.2.7_Setup.exe) was used and also require this software for viewing and analysis. 

Human subjects data

All human participants in the arm-based repellent efficacy assay against ticks provided explicit informed consent for their de-identified data to be published and shared in the public domain. This study was approved by the UC Santa Barbara Human Subjects Committee (Protocol #1-25-0697). No names, ages, sexes, or other personally identifiable or demographic information were collected or are included in the deposited dataset.