Data from: Intrinsic diversity in odor-evoked calcium rises across Drosophila olfactory neurons
Data files
Jul 29, 2026 version files 10.92 MB
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README.md
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Source_data_table_for_Figure_1_non_formatted.xlsx
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Source_data_table_for_Figure_1.xlsx
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Abstract
Dataset DOI: 10.5061/dryad.nvx0k6f74
Description of the data and file structure
These datasets include quantified spike and calcium data of Drosophila antennal olfactory receptor neurons through simultaneous recording, as well as quantified immunohistochemistry data from antennal specimen. The empty cells/columns contain no data and are required to improve readability of the data by serving as spacers. To improve accessibility of the dataset, non-formatted files are also provided, in which all formatting was removed and empty cells are filled with "nan".
Files and variables
File: Source_data_table_for_Figure_1.xlsx
Description: This Excel file contains source data for Figure 1. The workbook includes electrophysiological spike-frequency recordings and GCaMP calcium-imaging fluorescence responses from Drosophila olfactory receptor neurons in response to odor stimulation. Data are organized by olfactory neuron/receptor type in separate sheets, with individual fly measurements, average responses, standard error of the mean, and statistical analyses.
Sheets and variables
Sheet: ab2A_Or59b
Description: Electrophysiological and calcium-imaging responses of ab2A/Or59b neurons to methyl acetate stimulation.
Variables:
- Methyl acetate 10⁻⁷: Odor stimulus concentration used for the time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset. Negative values indicate pre-stimulus baseline; 0 indicates stimulus onset; positive values indicate post-stimulus response.
- Fly1–Fly7: Independent biological replicates (individual flies).
- Spike frequency (Hz): Electrophysiological firing rate measured in spikes per second.
- GCaMP fluorescence signal over time (ΔF/F, %): Calcium fluorescence response measured as percent change relative to baseline fluorescence.
- Methyl acetate 10⁻9 to 10⁻⁴: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response at each odor concentration.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response at each odor concentration.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Sheet: ab5A_Or82a
Description: Electrophysiological and calcium-imaging responses of ab5A/Or82a neurons to geranyl acetate stimulation.
Variables:
- Geranyl acetate 10⁻³: Odor stimulus concentration used for the time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset.
- Fly1–Fly7: Independent biological replicates (individual flies).
- Spike frequency (Hz): Electrophysiological firing rate measured in spikes per second.
- GCaMP fluorescence signal over time (ΔF/F, %): Calcium fluorescence response measured as percent change relative to baseline fluorescence.
- Geranyl acetate 10⁻4 to 10⁻1: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response at each odor concentration.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response at each odor concentration.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Sheet: Statistics
Description: Statistical comparisons between electrophysiological and calcium-imaging responses across ORN types.
Variables: Section: Wilcoxon rank-sum test (two tailed)
- Group 1: First experimental group in the comparison.
- Group 2: Second experimental group in the comparison.
- P value: Statistical significance of the comparison.
Section: Linear regression
- R²: Coefficient of determination for the linear regression between spike and calcium responses.
- Linear relationship: Regression equation describing the relationship between spike response (x, Hz) and calcium response (y, ΔF/F %).
Section: One-way MANOVA statistics
- Group 1: First ORN response profile.
- Group 2: Second ORN response profile.
- P value: Statistical significance of the multivariate comparison.
File: Source_data_table_for_Figure_3.xlsx
Description: This Excel file contains source data for Figure 3. The workbook compiles paired electrophysiological and calcium-imaging response measurements from 18 types of Drosophila olfactory receptor neurons (ORNs). Each sheet corresponds to one ORN type and includes peak odor-evoked spike responses and peak calcium fluorescence responses across multiple odor concentrations. The file also includes linear regression analyses quantifying the relationship between electrophysiological and calcium-imaging responses for each ORN type.
Sheets and variables
Sheet: Regression line equations
Description: Summary of linear regression analyses for all ORN types.
Variables:
- ORN type: Name of the olfactory receptor neuron and receptor identity (for example, ab2A/Or59b, ab2B/Or85a).
- Linear relationship: Best-fit regression equation relating calcium fluorescence response (y, ΔF/F %) to spike response (x, Hz).
- Coefficient of determination (R²): Measure of the goodness of fit for the linear regression.
- P value: Statistical significance of the linear regression.
Sheets: ab2A_Or59b, ab2B_Or85a, ab3A_Or22a, ab3B_Or85b, ab4A_Or7a, ab4B_Or56a, ab5A_Or82a, ab5B_Or47a, ac1A_Rh50, ac3A_Ir75b, ac3B_Or35a, ac4A_Ir84a, ac4C_Ir76a, ai3A_Or19a, ai3C_Or43a, at1_Or67d, at4A_Or47b, at4C_Or88a
Description: Each sheet contains paired peak spike and calcium responses for one ORN type across multiple odor concentrations.
Variables (common structure across ORN sheets):
Section: Peak spike response (Hz)
- Peak spike response (Hz): Maximum odor-evoked action potential firing rate measured by single-sensillum recording (SSR).
- [Odor-specific concentration column]: Concentration series for the cognate ligand used for that ORN. Examples include:
- methyl acetate (ab2A_Or59b)
- Ethyl 3-hydroxybutyrate (ab2B_Or85a)
- ethyl hexanoate (ab3A_Or22a)
- 2-heptanone (ab3B_Or85b)
- and other odor-specific ligands matching the sheet identity.
- Fly1–Fly7: Independent biological replicates.
- AVE: Mean peak spike response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Section: Peak Ca2+ response (ΔF/F, %)
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response measured as percent change relative to baseline fluorescence.
- [Same odor-specific concentration column]: Same concentration series used for the paired spike-response measurements.
- Fly1–Fly7: Independent biological replicates.
- AVE: Mean peak calcium response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
File: Source_data_table_for_Figure_4.xlsx
Description: This Excel file contains source data for Figure 4. The workbook includes quantitative morphological and physiological parameters of multiple Drosophila olfactory receptor neuron (ORN) types, along with correlation and statistical analyses. The morphological data was from Nava Gonzales et al., 2021.
Sheets and variables
Sheet: Correlation parameters
Description: Morphological and physiological measurements for each ORN type used in correlation analyses.
Variables:
- ORN type: Name of the olfactory receptor neuron and receptor identity (for example, ab2A/Or59b, ab3A/Or22a).
- Regression slope: Slope of the linear regression between peak spike response (Hz) and peak calcium response (ΔF/F %) for each ORN type. This value quantifies spike–calcium coupling sensitivity.
- Somatic surface area (µm²): Surface area of the neuronal soma.
- Somatic volume (µm³): Volume of the neuronal soma.
- Outer dendritic surface area (µm²): Surface area of the outer dendritic compartment exposed to odor stimuli.
- Outer dendritic volume (µm³): Volume of the outer dendritic compartment.
- Inner dendritic surface area (µm²): Surface area of the inner dendritic compartment.
- Inner dendritic volume (µm³): Volume of the inner dendritic compartment.
- Basal firing (Hz): Spontaneous firing rate measured before odor stimulation.
- Basal GCaMP fluorescence (AU): Baseline calcium fluorescence intensity measured in arbitrary units (AU).
- NA / NA*: Indicates data not available for that measurement.
- 132.79: Value marked with ** indicates acquisition using a different parameter, as annotated in the file.
Sheet: Statistics
Description: Statistical analyses of correlations between spike–calcium coupling and morphological or physiological parameters, as well as paired ORN comparisons.
Section: Pearson correlation analysis (Fig. 4b,c,g)
Variables:
- Regression slope vs outer dendritic surface area: Correlation between spike–calcium coupling slope and outer dendritic surface area.
- Regression slope vs outer dendritic volume: Correlation between spike–calcium coupling slope and outer dendritic volume.
- Regression slope vs inner dendritic surface area: Correlation between spike–calcium coupling slope and inner dendritic surface area.
- Regression slope vs inner dendritic volume: Correlation between spike–calcium coupling slope and inner dendritic volume.
- Regression slope vs somatic surface area: Correlation between spike–calcium coupling slope and somatic surface area.
- Regression slope vs somatic volume: Correlation between spike–calcium coupling slope and somatic volume.
- Regression slope vs basal firing: Correlation between spike–calcium coupling slope and spontaneous firing rate.
- Basal GCaMP fluorescence vs basal firing: Correlation between baseline fluorescence intensity and spontaneous firing rate.
- R: Pearson correlation coefficient.
- P value: Statistical significance of the correlation.
Section: One-way MANOVA statistics (Fig. 4d)
Variables:
- Group 1: First ORN type in the comparison.
- Group 2: Second ORN type in the comparison.
- P value: Statistical significance of the multivariate comparison.
File: Source_data_table_for_Figure_2.xlsx
Description: This Excel file contains source data for Figure 2. The workbook includes paired calcium-imaging and electrophysiological recordings from 18 olfactory receptor neuron (ORN) types in Drosophila antennae. Each sheet corresponds to one ORN type and contains odor-evoked responses measured across a range of odor concentrations. Data include GCaMP6m fluorescence traces and single-sensillum recording (SSR) spike-frequency measurements from individual flies.
Sheets and variables
Sheets: ab2A_Or59b, ab2B_Or85a, ab3A_Or22a, ab3B_Or85b, ab4A_Or7a, ab4B_Or56a, ab5A_Or82a, ab5B_Or47a, ac1A_Rh50, ac3A_Ir75b, ac3B_Or35a, ac4A_Ir84a, ac4C_Ir76a, ai3A_Or19a, ai3C_Or43a, at1_Or67d, at4A_Or47b, at4C_Or88a
Description: Each sheet contains calcium-imaging and single-sensillum recording (SSR) data for one ORN type.
Variables (common structure across sheets):
Calcium-imaging section:
- [Odor name]: Odor stimulus concentration column. The odor name differs by sheet and matches the receptor’s cognate ligand (examples include methyl acetate, ethyl 3-hydroxybutyrate, ethyl hexanoate, 2-heptanone, E2-hexenal, geosmin, geranyl acetate, etc). Values are dilutions (e.g., 10⁻⁶ dilution).
- frame: Imaging frame number. Calcium imaging was acquired at 10 Hz.
- Fly1–Fly7 (or Fly1–Fly8): Independent biological replicates.
- ΔF/F (%): Calcium fluorescence response expressed as percent change relative to baseline fluorescence.
- Imaging frame rate: 10 Hz: Acquisition rate for fluorescence recordings.
- Odor stimulus trigger: 68th frame: Frame at which odor stimulation begins.
Electrophysiology (SSR) section:
- Single-senillum recording: Electrophysiological recordings collected from the same ORN type.
- PSTH binned at 50 ms: Peri-stimulus time histogram data, grouped into 50 ms time bins.
- [Odor name]: Same odor concentration series used for the paired SSR recordings.
- frame: Time bin number corresponding to the PSTH.
- Fly1–Fly7 (or Fly1–Fly8): Independent biological replicates.
- Hz: Spike frequency measured in spikes per second.
Timing annotations:
- 0.3 sec pre-stimulus trigger: Baseline recording period before odor delivery.
- 1.9 sec post-stimulus trigger: Recording period after odor delivery.
File: Source_data_table_for_Figure_S1.xlsx
Description: This Excel file contains source data for Supplementary Figure S1. The workbook includes quantification of GFP fluorescence intensity driven by each Or-GAL4 driver in individual olfactory receptor neurons (ORNs) across multiple antennae in Drosophila.
Sheets and variables
Sheet: ab2A_Or59b
Description: Raw GFP fluorescence intensity measurements from ab2A/Or59b neurons.
Variables:
- Cell1–CellN: Individual olfactory receptor neurons measured within each antenna.
- Antenna1–Antenna9: Independent biological replicates (antenna preparations).
- GFP fluorescence (AU): Fluorescence intensity measured in arbitrary units (AU).
Sheet: ab3A_Or22a
Description: Raw GFP fluorescence intensity measurements from ab3A/Or22a neurons.
Variables:
- Cell1–CellN
- Antenna1–Antenna9
- GFP fluorescence (AU)
Sheet: ab5A_Or82a
Description: Raw GFP fluorescence intensity measurements from ab5A/Or82a neurons.
Variables:
- Cell1–CellN
- Antenna1–Antenna9
- GFP fluorescence (AU)
Sheet: ai3C_Or43a
Description: Raw GFP fluorescence intensity measurements from ai3C/Or43a neurons.
Variables:
- Cell1–CellN
- Antenna1–Antenna9
- GFP fluorescence (AU)
Sheet: Summary
Description: Mean GFP fluorescence per cell averaged across antennae for each ORN type.
Variables:
- Average fluorescence per cell: Label indicating that the values are average fluorescence intensity per cell.
- ab2A: Mean GFP fluorescence intensity per cell for ab2A neurons.
- ab3A: Mean GFP fluorescence intensity per cell for ab3A neurons.
- ab5A: Mean GFP fluorescence intensity per cell for ab5A neurons.
- ai3C: Mean GFP fluorescence intensity per cell for ai3C neurons.
- Antenna1–Antenna9: Individual biological replicates.
- AVG: Mean fluorescence across all antennae.
- SEM: Standard error of the mean across antennae.
Sheet: Statistics
Description: Statistical comparisons of GFP fluorescence between ORN types and correlation analysis with spike–calcium response relationship.
Section: One-way ANOVA
Variables:
- Group 1: First ORN type in the comparison.
- Group 2: Second ORN type in the comparison.
- P value: Statistical significance of the fluorescence difference between the two ORN groups.
Section: Pearson correlation analysis
Variables:
- Regression slope vs GFP fluorescence per cell: Correlation between spike–calcium response relationship slope (from Figure 3) and average GFP fluorescence per cell.
- R²: Coefficient of determination describing the strength of the correlation.
- P: Statistical significance of the correlation.
File: Source_data_table_for_Figure_5.xlsx
Description: This Excel file contains source data for Figure 5. The workbook includes electrophysiological and calcium-imaging measurements from Drosophila olfactory receptor neurons expressing either the endogenous receptor (Or22a or Or43a) or ectopically expressing both receptors (Or22a and Or43a). The data were used to assess how receptor co-expression alters odor tuning, basal firing activity, and the relationship between neuronal spiking and calcium responses.
Sheets and variables
Sheet: ab3A_Or22a
Description: Electrophysiological and calcium-imaging responses of ab3A neurons expressing endogenous Or22a alone.
Variables:
Section: Time-course recordings
- Ethyl hexanoate 10⁻7 and 1-methylindole 10-1: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset. Negative values indicate pre-stimulus; 0 indicates odor onset; positive values indicate post-stimulus.
- Fly1–Fly7: Independent biological replicates.
- Spike frequency (Hz): Electrophysiological firing rate measured in spikes per second.
- GCaMP fluorescence signal over time (ΔF/F, %): Calcium fluorescence response measured as percent change relative to baseline.
Section: Dose–response
- Ethyl hexanoate 10⁻9 to 10⁻4: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response at each concentration.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium response at each concentration.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean.
Sheet: ai3C_Or43a
Description: Electrophysiological and calcium-imaging responses of ai3C neurons expressing endogenous Or43a alone.
Variables:
Section: Time-course recordings
- Ethyl hexanoate 10⁻7 and 1-methylindole 10-1: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms)
- Fly1–Fly6
- Spike frequency (Hz)
- GCaMP fluorescence signal over time (ΔF/F, %)
Section: Dose–response
- 1-methylindole 10⁻3 to 3x10⁻1: Odor concentration series.
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: ab3A_Or22a_Or43a
Description: Electrophysiological and calcium-imaging responses of ab3A neurons co-expressing Or22a and Or43a.
Variables:
Section: Time-course recordings
- Ethyl hexanoate 10⁻6 and 1-methylindole 3x10-2: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms)
- Fly1–Fly6
- Spike frequency (Hz)
- GCaMP fluorescence signal over time (ΔF/F, %)
Section: Dose–response
- Ethyl-hexanoate 10-8 to 10-5 and 1-methylindole 10⁻3 to 10⁻1: Odor stimulus concentration used for time-course recordings.
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: Basal spike rate
Description: Baseline spontaneous firing rates of neurons before odor stimulation.
Variables:
- ab3A: Or22a: Basal spontaneous firing rate (Hz) for neurons expressing endogenous Or22a alone.
- ab3A: Or22a, Or43a: Basal spontaneous firing rate (Hz) for neurons co-expressing Or22a and Or43a.
- Neuron1–Neuron20: Individual biological replicates.
- AVE: Mean basal firing rate.
- SEM: Standard error of the mean.
Sheet: Statistics
Description: Statistical analyses comparing receptor-expression conditions.
Section: Linear regression
Variables:
- Experimental group: ORN/receptor condition analyzed.
- R²: Coefficient of determination for the linear regression between spike and calcium responses.
- Linear relationship: Best-fit regression equation relating calcium response (y, ΔF/F %) to spike response (x, Hz).
- P value: Statistical significance of the regression.
Section: One-way MANOVA statistics
Variables:
- Group 1: First response profile in the comparison.
- Group 2: Second response profile in the comparison.
- P value: Statistical significance of the multivariate comparison.
Section: Wilcoxon rank-sum test
Variables:
- Group 1: First basal firing group.
- Group 2: Second basal firing group.
- P value: Statistical significance of the basal firing comparison.
File: Source_data_table_for_Figure_S5.xlsx
Description: This Excel file contains source data for Supplementary Figure S5. The workbook includes quantitative measurements of basal fluorescence intensity, absolute odor-evoked fluorescence change (ΔF), and fluorescence responses normalized to basal fluorescence (ΔF/F) in Drosophila olfactory receptor neurons (ab2A/Or59b) following RNAi-mediated knockdown of intracellular calcium signaling and voltage-gated calcium channel components.
Sheets and variables
Sheet: Basal fluorescence
Description: Baseline GCaMP fluorescence intensity measured before odor stimulation.
Variables:
- Control (Ctrl): Baseline fluorescence values from genetic control flies.
- RyR RNAi KD: Baseline fluorescence values after ryanodine receptor (RyR) knockdown.
- IP3R RNAi KD: Baseline fluorescence values after inositol 1,4,5-trisphosphate receptor (IP3R) knockdown.
- Ca-α1D RNAi KD: Baseline fluorescence values after Ca-α1D channel knockdown.
- Ca-β RNAi KD: Baseline fluorescence values after Ca-β channel knockdown.
- Cacophony RNAi KD: Baseline fluorescence values after Cacophony channel knockdown.
- Fly1–Fly7: Independent biological replicates.
- Avg: Mean baseline fluorescence across biological replicates.
- SEM: Standard error of the mean across biological replicates.
- Basal fluorescence (AU): Fluorescence intensity reported in arbitrary units (AU).
Sheet: ΔF
Description: Absolute odor-evoked fluorescence changes.
Variables:
- Control (Ctrl): Absolute fluorescence change values from control flies.
- RyR RNAi KD
- IP3R RNAi KD
- Ca-α1D RNAi KD
- Ca-β RNAi KD
- Cacophony RNAi KD
- Fly1–Fly7
- Avg
- SEM
- ΔF (AU): Absolute fluorescence change calculated as peak fluorescence minus baseline fluorescence.
Sheet: ΔF normalized to basal F
Description: Odor-evoked fluorescence responses normalized to baseline fluorescence.
Variables:
- Control (Ctrl): Normalized fluorescence responses from control flies.
- RyR RNAi KD
- IP3R RNAi KD
- Ca-α1D RNAi KD
- Ca-β RNAi KD
- Cacophony RNAi KD
- Fly1–Fly7
- Avg
- SEM
- ΔF/F (%): Relative fluorescence change normalized to baseline fluorescence.
Sheet: Statistics
Description: Statistical comparisons of fluorescence measurements between control and RNAi knockdown conditions.
Section: Wilcoxon rank-sum test (two-tailed)
Variables:
- Measurement type: Indicates which fluorescence metric was tested (Basal fluorescence, ΔF, or ΔF/F).
- Group 1: Control group.
- Group 2: RNAi knockdown group.
- P value: Statistical significance of the comparison.
File: Source_data_table_for_Figure_S2.xlsx
Description: This Excel file contains source data for Supplementary Figure S2. The workbook includes odor-evoked calcium-imaging responses from multiple Drosophila olfactory receptor neuron (ORN) types expressing a different GCaMP variant GCaMP7c. These data are compared with previously measured GCaMP6m responses to evaluate the dynamic range and signal amplitude of the two calcium indicators across diverse ORN classes.
Sheets and variables
Sheets: ab2A_Or59b, ab2B_Or85a, ab3A_Or22a, ab3B_Or85b, ab4A_Or7a, ab4B_Or56a, ab5A_Or82a, ab5B_Or47a, ac1A_Rh50, ac3A_Ir75b, ac3B_Or35a, ac4A_Ir84a, ac4C_Ir76a, ai3A_Or19a, ai3C_Or43a, at1_Or67d, at4A_Or47b, at4C_Or88a
Description: Each sheet contains odor-evoked calcium-imaging recordings from one ORN type.
Variables (common structure across ORN sheets):
- [Odor-specific concentration column]: Odor stimulus concentration column. The exact odor is listed as the column header in each sheet. Examples include:
- methyl acetate (ab2A_Or59b)
- Ethyl 3-hydroxybutyrate (ab2B_Or85a)
- ethyl hexanoate (ab3A_Or22a)
- 2-heptanone (ab3B_Or85b)
- E2-hexenal (ab4A_Or7a)
- geosmin (ab4B_Or56a)
- geranyl acetate (ab5A_Or82a)
- pentyl acetate (ab5B_Or47a)
- ammonia (ac1A_Rh50)
- butyraldehyde (ac3A_Ir75b)
- 1-hexanol (ac3B_Or35a)
- phenylacetaldehyde (ac4A_Ir84a)
- phenylethylamine (ac4C_Ir76a)
- (S)-limonene (ai3A_Or19a)
- 1-methylindole (ai3C_Or43a)
- cis-vaccenyl acetate (at1_Or67d)
- palmitoleic acid (at4A_Or47b)
- methyl palmitate (at4C_Or88a)
- frame: Imaging frame number. Calcium imaging was acquired at 10 Hz.
- Fly1–Fly8: Independent biological replicates (number varies by sheet).
- ΔF/F (%): Calcium fluorescence response expressed as percent change relative to baseline fluorescence.
- Odor stimulus trigger (frame 70): Frame at which odor stimulation begins.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response at each concentration.
- AVE: Mean peak calcium response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Sheet: Highest Ca2+ response
Description: Summary comparison of the highest calcium responses measured using GCaMP6m and GCaMP7c across ORN types.
Variables:
- ORN type: Name of the olfactory receptor neuron class.
- GCaMP6m: Maximum calcium fluorescence response (ΔF/F, %) measured with GCaMP6m.
- GCaMP7c: Maximum calcium fluorescence response (ΔF/F, %) measured with GCaMP7c.
File: Source_data_table_for_Figure_S6.xlsx
Description: This Excel file contains source data for Supplementary Figure S6. The workbook summarizes the relationship between spike-calcium coupling (quantified as the regression slope between electrophysiological spike output and calcium-imaging response) and the endogenous transcript abundance of calcium channels across multiple Drosophila olfactory receptor neuron (ORN) types. Transcript abundance values were derived from published single-cell RNA sequencing datasets McLaughlin et al 2021.
Sheets and variables
Sheet: Sheet 1
Description: Quantitative comparison of spike–calcium coupling and calcium-channel transcript abundance across ORN types.
Variables:
- ORN type: Name of the olfactory receptor neuron class (for example, ab2A, ab3A, at1, ac3B).
- Regression slope: Slope of the linear regression between peak spike response (Hz) and peak calcium response (ΔF/F %) for that ORN type. This value reflects the sensitivity of calcium fluorescence changes relative to spike output.
- RyR transcript (log2(CPM+1)): Mean transcript abundance of the ryanodine receptor (RyR).
- IP3R transcript (log2(CPM+1)): Mean transcript abundance of the inositol 1,4,5-trisphosphate receptor (IP3R).
- Ca-α1D transcript (log2(CPM+1)): Mean transcript abundance of the voltage-gated calcium channel α1D subunit.
Section: Correlation analysis
Description: Pearson correlation analyses between regression slope and transcript abundance.
Variables:
- Channel: Calcium channel gene included in the correlation (RyR, IP3R, or Ca-α1D).
- R: Pearson correlation coefficient between transcript abundance and regression slope.
- P value: Statistical significance of the correlation.
Section: Transcript abundance definitions
Description: Criteria used for quantification of transcript values, as written in text.
Section: Criteria for defining ORN types
Description: Thresholds used to define ORN identity from the published single-cell RNA-seq dataset, as written in text.
File: Source_data_table_for_Figure_6.xlsx
Description: This Excel file contains source data for Figure 6. The workbook includes electrophysiological and calcium-imaging measurements from Drosophila olfactory receptor neurons (ORNs) following RNA interference (RNAi)-mediated knockdown of intracellular calcium and voltage-gated calcium channel components.
Sheets and variables
Sheet: RyR KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after ryanodine receptor (RyR) knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response measured by single-sensillum recording (SSR).
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response.
- Control (Ctrl): Responses from genetic control flies.
- RyR RNAi KD: Responses after RyR knockdown.
- Fly1–Fly7: Independent biological replicates.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean.
Sheet: IP3R KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after inositol 1,4,5-trisphosphate receptor (IP3R) knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- IP3R RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: Ca-α1D KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after Ca-α1D voltage-gated calcium channel knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- Ca-α1D RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: Ca-β KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after Ca-β subunit knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- Ca-β RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: Cacophony KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after Cacophony voltage-gated calcium channel knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- Cacophony RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: RyR KD in other ORNs
Description: Calcium-imaging responses in additional ORN types after RyR knockdown.
Variables:
- Ethyl 3-hydroxybutyrate: Odor stimulus used for ab2B/Or85a neurons.
- Ethyl hexanoate: Odor stimulus used for ab3A/Or22a neurons.
- Ctrl: Control calcium responses.
- RyR RNAi KD: Calcium responses after RyR knockdown.
- Fly1–Fly7: Independent biological replicates.
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: IP3R KD in other ORNs
Description: Calcium-imaging responses in additional ORN types after IP3R knockdown.
Variables:
- Ethyl 3-hydroxybutyrate
- Ethyl hexanoate
- Ctrl
- IP3R RNAi KD
- Fly1–Fly7
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: Ca-α1D KD in other ORNs
Description: Calcium-imaging responses in additional ORN types after Ca-α1D knockdown.
Variables:
- Ethyl 3-hydroxybutyrate
- Ethyl hexanoate
- Ctrl
- Ca-α1D RNAi KD
- Fly1–Fly7
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: Statistics
Description: Statistical analyses of RNAi knockdown effects.
Section: Linear regression
Variables:
- Experimental group: RNAi condition analyzed.
- R²: Coefficient of determination for the linear regression between spike and calcium responses.
- Linear relationship: Best-fit regression equation relating calcium response (y, ΔF/F %) to spike response (x, Hz).
- P value: Statistical significance of the regression.
Section: One-way MANOVA statistics
Variables:
- Group 1: First genotype/condition in the comparison.
- Group 2: Second genotype/condition in the comparison.
- P value: Statistical significance of the multivariate comparison.
Section: Wilcoxon rank-sum test
Variables:
- Group 1: Control group.
- Group 2: RNAi knockdown group.
- P value: Statistical significance of the comparison.
File: Source_data_table_for_Figure_S3.xlsx
Description: This Excel file contains source data for Supplementary Figure S3. The workbook includes odor-evoked local field potential (LFP) recordings, spike-frequency measurements, and basal firing rates from Drosophila olfactory receptor neurons under two experimental conditions: without background odor stimulation and with continuous background odor stimulation.
Sheets and variables
Sheet: ab3A
Description: Electrophysiological responses of ab3A/Or22a neurons to ethyl hexanoate with and without background odor stimulation.
Variables:
Section: Without background odor stimulation
- Ethyl hexanoate 10⁻7: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset.
- Fly1–Fly7: Independent biological replicates.
- LFP (mV): Local field potential response measured in millivolts.
- Spike frequency (Hz): Odor-evoked firing rate measured in spikes per second.
Section: With background odor stimulation
- Ethyl hexanoate 10⁻7 + background odor: Same odor stimulus delivered during continuous background odor exposure.
- Time points relative to stimulus onset (ms)
- Fly1–Fly7
- LFP (mV)
- Spike frequency (Hz)
Section: Basal firing rate
- Basal firing (Hz): Baseline firing rate before odor stimulus.
- Fly1–Fly7: Individual biological replicates.
- AVE: Mean basal firing rate.
- SEM: Standard error of the mean.
Section: Dose–response sensitivity
- Ethyl hexanoate 10⁻9 to 3x10⁻5: Odor concentration series used for pEC50 calculations.
- LFP pEC50: Negative logarithm of the odor concentration producing half-maximal LFP response.
- Spike pEC50: Negative logarithm of the odor concentration producing half-maximal spike response.
Sheet: ai3A
Description: Electrophysiological responses of ai3A/Or43a neurons to (S)-limonene with and without background odor stimulation.
Variables:
Section: Without background odor stimulation
- (S)-limonene 3x10⁻3: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms)
- Fly1–Fly6: Independent biological replicates.
- LFP (mV)
- Spike frequency (Hz)
Section: With background odor stimulation
- (S)-limonene 3x10⁻3 + background odor
- Time points relative to stimulus onset (ms)
- Fly1–Fly6
- LFP (mV)
- Spike frequency (Hz)
Section: Basal firing rate
- Basal firing (Hz)
- Fly1–Fly6
- AVE
- SEM
Section: Dose–response sensitivity
- (S)-limonene 10⁻3 to 10⁻¹: Odor concentration series used for pEC50 calculations.
- LFP pEC50
- Spike pEC50
Sheet: Statistics
Description: Statistical comparisons of basal firing and odor sensitivity with and without background odor stimulation.
Section: Wilcoxon rank-sum test (two-tailed)
Variables:
- Group 1: Condition without background odor.
- Group 2: Condition with background odor.
- P value: Statistical significance of the comparison.
Comparisons include:
- Basal firing rate (Hz)
- LFP pEC50
- Spike pEC50
File: Source_data_table_for_Figure_1_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Figure 1. The workbook includes electrophysiological spike-frequency recordings and GCaMP calcium-imaging fluorescence responses from Drosophila olfactory receptor neurons in response to odor stimulation. Data are organized by olfactory neuron/receptor type in separate sheets, with individual fly measurements, average responses, standard error of the mean, and statistical analyses.
Sheets and variables
Sheet: ab2A_Or59b
Description: Electrophysiological and calcium-imaging responses of ab2A/Or59b neurons to methyl acetate stimulation.
Variables:
- Methyl acetate 10⁻⁷: Odor stimulus concentration used for the time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset. Negative values indicate pre-stimulus baseline; 0 indicates stimulus onset; positive values indicate post-stimulus response.
- Fly1–Fly7: Independent biological replicates (individual flies).
- Spike frequency (Hz): Electrophysiological firing rate measured in spikes per second.
- GCaMP fluorescence signal over time (ΔF/F, %): Calcium fluorescence response measured as percent change relative to baseline fluorescence.
- Methyl acetate 10⁻9 to 10⁻⁴: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response at each odor concentration.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response at each odor concentration.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Sheet: ab5A_Or82a
Description: Electrophysiological and calcium-imaging responses of ab5A/Or82a neurons to geranyl acetate stimulation.
Variables:
- Geranyl acetate 10⁻³: Odor stimulus concentration used for the time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset.
- Fly1–Fly7: Independent biological replicates (individual flies).
- Spike frequency (Hz): Electrophysiological firing rate measured in spikes per second.
- GCaMP fluorescence signal over time (ΔF/F, %): Calcium fluorescence response measured as percent change relative to baseline fluorescence.
- Geranyl acetate 10⁻4 to 10⁻1: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response at each odor concentration.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response at each odor concentration.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Sheet: Statistics
Description: Statistical comparisons between electrophysiological and calcium-imaging responses across ORN types.
Variables:
Section: Wilcoxon rank-sum test (two tailed)
- Group 1: First experimental group in the comparison.
- Group 2: Second experimental group in the comparison.
- P value: Statistical significance of the comparison.
Section: Linear regression
- R²: Coefficient of determination for the linear regression between spike and calcium responses.
- Linear relationship: Regression equation describing the relationship between spike response (x, Hz) and calcium response (y, ΔF/F %).
Section: One-way MANOVA statistics
- Group 1: First ORN response profile.
- Group 2: Second ORN response profile.
- P value: Statistical significance of the multivariate comparison.
File: Source_data_table_for_Figure_3_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Figure 3. The workbook compiles paired electrophysiological and calcium-imaging response measurements from 18 types of Drosophila olfactory receptor neurons (ORNs). Each sheet corresponds to one ORN type and includes peak odor-evoked spike responses and peak calcium fluorescence responses across multiple odor concentrations. The file also includes linear regression analyses quantifying the relationship between electrophysiological and calcium-imaging responses for each ORN type.
Sheets and variables
Sheet: Regression line equations
Description: Summary of linear regression analyses for all ORN types.
Variables:
- ORN type: Name of the olfactory receptor neuron and receptor identity (for example, ab2A/Or59b, ab2B/Or85a).
- Linear relationship: Best-fit regression equation relating calcium fluorescence response (y, ΔF/F %) to spike response (x, Hz).
- Coefficient of determination (R²): Measure of the goodness of fit for the linear regression.
- P value: Statistical significance of the linear regression.
Sheets: ab2A_Or59b, ab2B_Or85a, ab3A_Or22a, ab3B_Or85b, ab4A_Or7a, ab4B_Or56a, ab5A_Or82a, ab5B_Or47a, ac1A_Rh50, ac3A_Ir75b, ac3B_Or35a, ac4A_Ir84a, ac4C_Ir76a, ai3A_Or19a, ai3C_Or43a, at1_Or67d, at4A_Or47b, at4C_Or88a
Description: Each sheet contains paired peak spike and calcium responses for one ORN type across multiple odor concentrations.
Variables (common structure across ORN sheets):
Section: Peak spike response (Hz)
- Peak spike response (Hz): Maximum odor-evoked action potential firing rate measured by single-sensillum recording (SSR).
- [Odor-specific concentration column]: Concentration series for the cognate ligand used for that ORN. Examples include:
- methyl acetate (ab2A_Or59b)
- Ethyl 3-hydroxybutyrate (ab2B_Or85a)
- ethyl hexanoate (ab3A_Or22a)
- 2-heptanone (ab3B_Or85b)
- and other odor-specific ligands matching the sheet identity.
- Fly1–Fly7: Independent biological replicates.
- AVE: Mean peak spike response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Section: Peak Ca2+ response (ΔF/F, %)
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response measured as percent change relative to baseline fluorescence.
- [Same odor-specific concentration column]: Same concentration series used for the paired spike-response measurements.
- Fly1–Fly7: Independent biological replicates.
- AVE: Mean peak calcium response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
File: Source_data_table_for_Figure_4_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Figure 4. The workbook includes quantitative morphological and physiological parameters of multiple Drosophila olfactory receptor neuron (ORN) types, along with correlation and statistical analyses. The morphological data was from Nava Gonzales et al., 2021.
Sheets and variables
Sheet: Correlation parameters
Description: Morphological and physiological measurements for each ORN type used in correlation analyses.
Variables:
- ORN type: Name of the olfactory receptor neuron and receptor identity (for example, ab2A/Or59b, ab3A/Or22a).
- Regression slope: Slope of the linear regression between peak spike response (Hz) and peak calcium response (ΔF/F %) for each ORN type. This value quantifies spike–calcium coupling sensitivity.
- Somatic surface area (µm²): Surface area of the neuronal soma.
- Somatic volume (µm³): Volume of the neuronal soma.
- Outer dendritic surface area (µm²): Surface area of the outer dendritic compartment exposed to odor stimuli.
- Outer dendritic volume (µm³): Volume of the outer dendritic compartment.
- Inner dendritic surface area (µm²): Surface area of the inner dendritic compartment.
- Inner dendritic volume (µm³): Volume of the inner dendritic compartment.
- Basal firing (Hz): Spontaneous firing rate measured before odor stimulation.
- Basal GCaMP fluorescence (AU): Baseline calcium fluorescence intensity measured in arbitrary units (AU).
- NA / NA*: Indicates data not available for that measurement.
- 132.79: Value marked with ** indicates acquisition using a different parameter, as annotated in the file.
Sheet: Statistics
Description: Statistical analyses of correlations between spike–calcium coupling and morphological or physiological parameters, as well as paired ORN comparisons.
Section: Pearson correlation analysis (Fig. 4b,c,g)
Variables:
- Regression slope vs outer dendritic surface area: Correlation between spike–calcium coupling slope and outer dendritic surface area.
- Regression slope vs outer dendritic volume: Correlation between spike–calcium coupling slope and outer dendritic volume.
- Regression slope vs inner dendritic surface area: Correlation between spike–calcium coupling slope and inner dendritic surface area.
- Regression slope vs inner dendritic volume: Correlation between spike–calcium coupling slope and inner dendritic volume.
- Regression slope vs somatic surface area: Correlation between spike–calcium coupling slope and somatic surface area.
- Regression slope vs somatic volume: Correlation between spike–calcium coupling slope and somatic volume.
- Regression slope vs basal firing: Correlation between spike–calcium coupling slope and spontaneous firing rate.
- Basal GCaMP fluorescence vs basal firing: Correlation between baseline fluorescence intensity and spontaneous firing rate.
- R: Pearson correlation coefficient.
- P value: Statistical significance of the correlation.
Section: One-way MANOVA statistics (Fig. 4d)
Variables:
- Group 1: First ORN type in the comparison.
- Group 2: Second ORN type in the comparison.
- P value: Statistical significance of the multivariate comparison.
File: Source_data_table_for_Figure_2_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Figure 2. The workbook includes paired calcium-imaging and electrophysiological recordings from 18 olfactory receptor neuron (ORN) types in Drosophila antennae. Each sheet corresponds to one ORN type and contains odor-evoked responses measured across a range of odor concentrations. Data include GCaMP6m fluorescence traces and single-sensillum recording (SSR) spike-frequency measurements from individual flies.
Sheets and variables
Sheets: ab2A_Or59b, ab2B_Or85a, ab3A_Or22a, ab3B_Or85b, ab4A_Or7a, ab4B_Or56a, ab5A_Or82a, ab5B_Or47a, ac1A_Rh50, ac3A_Ir75b, ac3B_Or35a, ac4A_Ir84a, ac4C_Ir76a, ai3A_Or19a, ai3C_Or43a, at1_Or67d, at4A_Or47b, at4C_Or88a
Description: Each sheet contains calcium-imaging and single-sensillum recording (SSR) data for one ORN type.
Variables (common structure across sheets):
**Calcium-imaging section:
**
- [Odor name]: Odor stimulus concentration column. The odor name differs by sheet and matches the receptor’s cognate ligand (examples include methyl acetate, ethyl 3-hydroxybutyrate, ethyl hexanoate, 2-heptanone, E2-hexenal, geosmin, geranyl acetate, etc). Values are dilutions (e.g., 10⁻⁶ dilution).
- frame: Imaging frame number. Calcium imaging was acquired at 10 Hz.
- Fly1–Fly7 (or Fly1–Fly8): Independent biological replicates.
- ΔF/F (%): Calcium fluorescence response expressed as percent change relative to baseline fluorescence.
- Imaging frame rate: 10 Hz: Acquisition rate for fluorescence recordings.
- Odor stimulus trigger: 68th frame: Frame at which odor stimulation begins.
**Electrophysiology (SSR) section:
**
- Single-senillum recording: Electrophysiological recordings collected from the same ORN type.
- PSTH binned at 50 ms: Peri-stimulus time histogram data, grouped into 50 ms time bins.
- [Odor name]: Same odor concentration series used for the paired SSR recordings.
- frame: Time bin number corresponding to the PSTH.
- Fly1–Fly7 (or Fly1–Fly8): Independent biological replicates.
- Hz: Spike frequency measured in spikes per second.
**Timing annotations:
**
- 0.3 sec pre-stimulus trigger: Baseline recording period before odor delivery.
- 1.9 sec post-stimulus trigger: Recording period after odor delivery.
File: Source_data_table_for_Figure_S1_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Supplementary Figure S1. The workbook includes quantification of GFP fluorescence intensity driven by each Or-GAL4 driver in individual olfactory receptor neurons (ORNs) across multiple antennae in Drosophila.
Sheets and variables
Sheet: ab2A_Or59b
Description: Raw GFP fluorescence intensity measurements from ab2A/Or59b neurons.
Variables:
- Cell1–CellN: Individual olfactory receptor neurons measured within each antenna.
- Antenna1–Antenna9: Independent biological replicates (antenna preparations).
- GFP fluorescence (AU): Fluorescence intensity measured in arbitrary units (AU).
Sheet: ab3A_Or22a
Description: Raw GFP fluorescence intensity measurements from ab3A/Or22a neurons.
Variables:
- Cell1–CellN
- Antenna1–Antenna9
- GFP fluorescence (AU)
Sheet: ab5A_Or82a
Description: Raw GFP fluorescence intensity measurements from ab5A/Or82a neurons.
Variables:
- Cell1–CellN
- Antenna1–Antenna9
- GFP fluorescence (AU)
Sheet: ai3C_Or43a
Description: Raw GFP fluorescence intensity measurements from ai3C/Or43a neurons.
Variables:
- Cell1–CellN
- Antenna1–Antenna9
- GFP fluorescence (AU)
Sheet: Summary
Description: Mean GFP fluorescence per cell averaged across antennae for each ORN type.
Variables:
- Average fluorescence per cell: Label indicating that the values are average fluorescence intensity per cell.
- ab2A: Mean GFP fluorescence intensity per cell for ab2A neurons.
- ab3A: Mean GFP fluorescence intensity per cell for ab3A neurons.
- ab5A: Mean GFP fluorescence intensity per cell for ab5A neurons.
- ai3C: Mean GFP fluorescence intensity per cell for ai3C neurons.
- Antenna1–Antenna9: Individual biological replicates.
- AVG: Mean fluorescence across all antennae.
- SEM: Standard error of the mean across antennae.
Sheet: Statistics
Description: Statistical comparisons of GFP fluorescence between ORN types and correlation analysis with spike–calcium response relationship.
Section: One-way ANOVA
Variables:
- Group 1: First ORN type in the comparison.
- Group 2: Second ORN type in the comparison.
- P value: Statistical significance of the fluorescence difference between the two ORN groups.
Section: Pearson correlation analysis
Variables:
- Regression slope vs GFP fluorescence per cell: Correlation between spike–calcium response relationship slope (from Figure 3) and average GFP fluorescence per cell.
- R²: Coefficient of determination describing the strength of the correlation.
- P: Statistical significance of the correlation.
File: Source_data_table_for_Figure_5_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Figure 5. The workbook includes electrophysiological and calcium-imaging measurements from Drosophila olfactory receptor neurons expressing either the endogenous receptor (Or22a or Or43a) or ectopically expressing both receptors (Or22a and Or43a). The data were used to assess how receptor co-expression alters odor tuning, basal firing activity, and the relationship between neuronal spiking and calcium responses.
Sheets and variables
Sheet: ab3A_Or22a
Description: Electrophysiological and calcium-imaging responses of ab3A neurons expressing endogenous Or22a alone.
Variables:
Section: Time-course recordings
- Ethyl hexanoate 10⁻7 and 1-methylindole 10-1: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset. Negative values indicate pre-stimulus; 0 indicates odor onset; positive values indicate post-stimulus.
- Fly1–Fly7: Independent biological replicates.
- Spike frequency (Hz): Electrophysiological firing rate measured in spikes per second.
- GCaMP fluorescence signal over time (ΔF/F, %): Calcium fluorescence response measured as percent change relative to baseline.
Section: Dose–response
- Ethyl hexanoate 10⁻9 to 10⁻4: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response at each concentration.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium response at each concentration.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean.
Sheet: ai3C_Or43a
Description: Electrophysiological and calcium-imaging responses of ai3C neurons expressing endogenous Or43a alone.
Variables:
Section: Time-course recordings
- Ethyl hexanoate 10⁻7 and 1-methylindole 10-1: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms)
- Fly1–Fly6
- Spike frequency (Hz)
- GCaMP fluorescence signal over time (ΔF/F, %)
Section: Dose–response
- 1-methylindole 10⁻3 to 3x10⁻1: Odor concentration series.
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: ab3A_Or22a_Or43a
Description: Electrophysiological and calcium-imaging responses of ab3A neurons co-expressing Or22a and Or43a.
Variables:
Section: Time-course recordings
- Ethyl hexanoate 10⁻6 and 1-methylindole 3x10-2: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms)
- Fly1–Fly6
- Spike frequency (Hz)
- GCaMP fluorescence signal over time (ΔF/F, %)
Section: Dose–response
- Ethyl-hexanoate 10-8 to 10-5 and 1-methylindole 10⁻3 to 10⁻1: Odor stimulus concentration used for time-course recordings.
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: Basal spike rate
Description: Baseline spontaneous firing rates of neurons before odor stimulation.
Variables:
- ab3A: Or22a: Basal spontaneous firing rate (Hz) for neurons expressing endogenous Or22a alone.
- ab3A: Or22a, Or43a: Basal spontaneous firing rate (Hz) for neurons co-expressing Or22a and Or43a.
- Neuron1–Neuron20: Individual biological replicates.
- AVE: Mean basal firing rate.
- SEM: Standard error of the mean.
Sheet: Statistics
Description: Statistical analyses comparing receptor-expression conditions.
Section: Linear regression
Variables:
- Experimental group: ORN/receptor condition analyzed.
- R²: Coefficient of determination for the linear regression between spike and calcium responses.
- Linear relationship: Best-fit regression equation relating calcium response (y, ΔF/F %) to spike response (x, Hz).
- P value: Statistical significance of the regression.
Section: One-way MANOVA statistics
Variables:
- Group 1: First response profile in the comparison.
- Group 2: Second response profile in the comparison.
- P value: Statistical significance of the multivariate comparison.
Section: Wilcoxon rank-sum test
Variables:
- Group 1: First basal firing group.
- Group 2: Second basal firing group.
- P value: Statistical significance of the basal firing comparison.
File: Source_data_table_for_Figure_S5_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Supplementary Figure S5. The workbook includes quantitative measurements of basal fluorescence intensity, absolute odor-evoked fluorescence change (ΔF), and fluorescence responses normalized to basal fluorescence (ΔF/F) in Drosophila olfactory receptor neurons (ab2A/Or59b) following RNAi-mediated knockdown of intracellular calcium signaling and voltage-gated calcium channel components.
Sheets and variables
Sheet: Basal fluorescence
Description: Baseline GCaMP fluorescence intensity measured before odor stimulation.
Variables:
- Control (Ctrl): Baseline fluorescence values from genetic control flies.
- RyR RNAi KD: Baseline fluorescence values after ryanodine receptor (RyR) knockdown.
- IP3R RNAi KD: Baseline fluorescence values after inositol 1,4,5-trisphosphate receptor (IP3R) knockdown.
- Ca-α1D RNAi KD: Baseline fluorescence values after Ca-α1D channel knockdown.
- Ca-β RNAi KD: Baseline fluorescence values after Ca-β channel knockdown.
- Cacophony RNAi KD: Baseline fluorescence values after Cacophony channel knockdown.
- Fly1–Fly7: Independent biological replicates.
- Avg: Mean baseline fluorescence across biological replicates.
- SEM: Standard error of the mean across biological replicates.
- Basal fluorescence (AU): Fluorescence intensity reported in arbitrary units (AU).
Sheet: ΔF
Description: Absolute odor-evoked fluorescence changes.
Variables:
- Control (Ctrl): Absolute fluorescence change values from control flies.
- RyR RNAi KD
- IP3R RNAi KD
- Ca-α1D RNAi KD
- Ca-β RNAi KD
- Cacophony RNAi KD
- Fly1–Fly7
- Avg
- SEM
- ΔF (AU): Absolute fluorescence change calculated as peak fluorescence minus baseline fluorescence.
Sheet: ΔF normalized to basal F
Description: Odor-evoked fluorescence responses normalized to baseline fluorescence.
Variables:
- Control (Ctrl): Normalized fluorescence responses from control flies.
- RyR RNAi KD
- IP3R RNAi KD
- Ca-α1D RNAi KD
- Ca-β RNAi KD
- Cacophony RNAi KD
- Fly1–Fly7
- Avg
- SEM
- ΔF/F (%): Relative fluorescence change normalized to baseline fluorescence.
Sheet: Statistics
Description: Statistical comparisons of fluorescence measurements between control and RNAi knockdown conditions.
Section: Wilcoxon rank-sum test (two-tailed)
Variables:
- Measurement type: Indicates which fluorescence metric was tested (Basal fluorescence, ΔF, or ΔF/F).
- Group 1: Control group.
- Group 2: RNAi knockdown group.
- P value: Statistical significance of the comparison.
File: Source_data_table_for_Figure_S2_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Supplementary Figure S2. The workbook includes odor-evoked calcium-imaging responses from multiple Drosophila olfactory receptor neuron (ORN) types expressing a different GCaMP variant GCaMP7c. These data are compared with previously measured GCaMP6m responses to evaluate the dynamic range and signal amplitude of the two calcium indicators across diverse ORN classes.
Sheets and variables
Sheets: ab2A_Or59b, ab2B_Or85a, ab3A_Or22a, ab3B_Or85b, ab4A_Or7a, ab4B_Or56a, ab5A_Or82a, ab5B_Or47a, ac1A_Rh50, ac3A_Ir75b, ac3B_Or35a, ac4A_Ir84a, ac4C_Ir76a, ai3A_Or19a, ai3C_Or43a, at1_Or67d, at4A_Or47b, at4C_Or88a
Description: Each sheet contains odor-evoked calcium-imaging recordings from one ORN type.
Variables (common structure across ORN sheets):
- [Odor-specific concentration column]: Odor stimulus concentration column. The exact odor is listed as the column header in each sheet. Examples include:
- methyl acetate (ab2A_Or59b)
- Ethyl 3-hydroxybutyrate (ab2B_Or85a)
- ethyl hexanoate (ab3A_Or22a)
- 2-heptanone (ab3B_Or85b)
- E2-hexenal (ab4A_Or7a)
- geosmin (ab4B_Or56a)
- geranyl acetate (ab5A_Or82a)
- pentyl acetate (ab5B_Or47a)
- ammonia (ac1A_Rh50)
- butyraldehyde (ac3A_Ir75b)
- 1-hexanol (ac3B_Or35a)
- phenylacetaldehyde (ac4A_Ir84a)
- phenylethylamine (ac4C_Ir76a)
- (S)-limonene (ai3A_Or19a)
- 1-methylindole (ai3C_Or43a)
- cis-vaccenyl acetate (at1_Or67d)
- palmitoleic acid (at4A_Or47b)
- methyl palmitate (at4C_Or88a)
- frame: Imaging frame number. Calcium imaging was acquired at 10 Hz.
- Fly1–Fly8: Independent biological replicates (number varies by sheet).
- ΔF/F (%): Calcium fluorescence response expressed as percent change relative to baseline fluorescence.
- Odor stimulus trigger (frame 70): Frame at which odor stimulation begins.
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response at each concentration.
- AVE: Mean peak calcium response across biological replicates.
- SEM: Standard error of the mean across biological replicates.
Sheet: Highest Ca2+ response
Description: Summary comparison of the highest calcium responses measured using GCaMP6m and GCaMP7c across ORN types.
Variables:
- ORN type: Name of the olfactory receptor neuron class.
- GCaMP6m: Maximum calcium fluorescence response (ΔF/F, %) measured with GCaMP6m.
- GCaMP7c: Maximum calcium fluorescence response (ΔF/F, %) measured with GCaMP7c.
File: Source_data_table_for_Figure_S6_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Supplementary Figure S6. The workbook summarizes the relationship between spike-calcium coupling (quantified as the regression slope between electrophysiological spike output and calcium-imaging response) and the endogenous transcript abundance of calcium channels across multiple Drosophila olfactory receptor neuron (ORN) types. Transcript abundance values were derived from published single-cell RNA sequencing datasets McLaughlin et al 2021.
Sheets and variables
Sheet: Sheet 1
Description: Quantitative comparison of spike–calcium coupling and calcium-channel transcript abundance across ORN types.
Variables:
- ORN type: Name of the olfactory receptor neuron class (for example, ab2A, ab3A, at1, ac3B).
- Regression slope: Slope of the linear regression between peak spike response (Hz) and peak calcium response (ΔF/F %) for that ORN type. This value reflects the sensitivity of calcium fluorescence changes relative to spike output.
- RyR transcript (log2(CPM+1)): Mean transcript abundance of the ryanodine receptor (RyR).
- IP3R transcript (log2(CPM+1)): Mean transcript abundance of the inositol 1,4,5-trisphosphate receptor (IP3R).
- Ca-α1D transcript (log2(CPM+1)): Mean transcript abundance of the voltage-gated calcium channel α1D subunit.
Section: Correlation analysis
Description: Pearson correlation analyses between regression slope and transcript abundance.
Variables:
- Channel: Calcium channel gene included in the correlation (RyR, IP3R, or Ca-α1D).
- R: Pearson correlation coefficient between transcript abundance and regression slope.
- P value: Statistical significance of the correlation.
Section: Transcript abundance definitions
Description: Criteria used for quantification of transcript values, as written in text.
Section: Criteria for defining ORN types
Description: Thresholds used to define ORN identity from the published single-cell RNA-seq dataset, as written in text.
File: Source_data_table_for_Figure_6_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Figure 6. The workbook includes electrophysiological and calcium-imaging measurements from Drosophila olfactory receptor neurons (ORNs) following RNA interference (RNAi)-mediated knockdown of intracellular calcium and voltage-gated calcium channel components.
Sheets and variables
Sheet: RyR KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after ryanodine receptor (RyR) knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴: Odor concentration series used for dose–response analysis.
- Peak spike response (Hz): Maximum odor-evoked spike response measured by single-sensillum recording (SSR).
- Peak Ca2+ response (ΔF/F, %): Maximum odor-evoked calcium fluorescence response.
- Control (Ctrl): Responses from genetic control flies.
- RyR RNAi KD: Responses after RyR knockdown.
- Fly1–Fly7: Independent biological replicates.
- AVE: Mean response across biological replicates.
- SEM: Standard error of the mean.
Sheet: IP3R KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after inositol 1,4,5-trisphosphate receptor (IP3R) knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- IP3R RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: Ca-α1D KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after Ca-α1D voltage-gated calcium channel knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- Ca-α1D RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: Ca-β KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after Ca-β subunit knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- Ca-β RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: Cacophony KD in ab2A_Or59b
Description: Responses of ab2A/Or59b neurons after Cacophony voltage-gated calcium channel knockdown.
Variables:
- Methyl acetate 10⁻9 to 10⁻⁴
- Peak spike response (Hz)
- Peak Ca2+ response (ΔF/F, %)
- Control (Ctrl)
- Cacophony RNAi KD
- Fly1–Fly7
- AVE
- SEM
Sheet: RyR KD in other ORNs
Description: Calcium-imaging responses in additional ORN types after RyR knockdown.
Variables:
- Ethyl 3-hydroxybutyrate: Odor stimulus used for ab2B/Or85a neurons.
- Ethyl hexanoate: Odor stimulus used for ab3A/Or22a neurons.
- Ctrl: Control calcium responses.
- RyR RNAi KD: Calcium responses after RyR knockdown.
- Fly1–Fly7: Independent biological replicates.
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: IP3R KD in other ORNs
Description: Calcium-imaging responses in additional ORN types after IP3R knockdown.
Variables:
- Ethyl 3-hydroxybutyrate
- Ethyl hexanoate
- Ctrl
- IP3R RNAi KD
- Fly1–Fly7
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: Ca-α1D KD in other ORNs
Description: Calcium-imaging responses in additional ORN types after Ca-α1D knockdown.
Variables:
- Ethyl 3-hydroxybutyrate
- Ethyl hexanoate
- Ctrl
- Ca-α1D RNAi KD
- Fly1–Fly7
- Peak Ca2+ response (ΔF/F, %)
- AVE
- SEM
Sheet: Statistics
Description: Statistical analyses of RNAi knockdown effects.
Section: Linear regression
Variables:
- Experimental group: RNAi condition analyzed.
- R²: Coefficient of determination for the linear regression between spike and calcium responses.
- Linear relationship: Best-fit regression equation relating calcium response (y, ΔF/F %) to spike response (x, Hz).
- P value: Statistical significance of the regression.
Section: One-way MANOVA statistics
Variables:
- Group 1: First genotype/condition in the comparison.
- Group 2: Second genotype/condition in the comparison.
- P value: Statistical significance of the multivariate comparison.
Section: Wilcoxon rank-sum test
Variables:
- Group 1: Control group.
- Group 2: RNAi knockdown group.
- P value: Statistical significance of the comparison.
File: Source_data_table_for_Figure_S3_non_formatted.xlsx
Description: This Excel file contains non-formatted source data for Supplementary Figure S3. The workbook includes odor-evoked local field potential (LFP) recordings, spike-frequency measurements, and basal firing rates from Drosophila olfactory receptor neurons under two experimental conditions: without background odor stimulation and with continuous background odor stimulation.
Sheets and variables
Sheet: ab3A
Description: Electrophysiological responses of ab3A/Or22a neurons to ethyl hexanoate with and without background odor stimulation.
Variables:
Section: Without background odor stimulation
- Ethyl hexanoate 10⁻7: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms): Time in milliseconds relative to odor onset.
- Fly1–Fly7: Independent biological replicates.
- LFP (mV): Local field potential response measured in millivolts.
- Spike frequency (Hz): Odor-evoked firing rate measured in spikes per second.
Section: With background odor stimulation
- Ethyl hexanoate 10⁻7 + background odor: Same odor stimulus delivered during continuous background odor exposure.
- Time points relative to stimulus onset (ms)
- Fly1–Fly7
- LFP (mV)
- Spike frequency (Hz)
Section: Basal firing rate
- Basal firing (Hz): Baseline firing rate before odor stimulus.
- Fly1–Fly7: Individual biological replicates.
- AVE: Mean basal firing rate.
- SEM: Standard error of the mean.
Section: Dose–response sensitivity
- Ethyl hexanoate 10⁻9 to 3x10⁻5: Odor concentration series used for pEC50 calculations.
- LFP pEC50: Negative logarithm of the odor concentration producing half-maximal LFP response.
- Spike pEC50: Negative logarithm of the odor concentration producing half-maximal spike response.
Sheet: ai3A
Description: Electrophysiological responses of ai3A/Or43a neurons to (S)-limonene with and without background odor stimulation.
Variables:
Section: Without background odor stimulation
- (S)-limonene 3x10⁻3: Odor stimulus concentration used for time-course recordings.
- Time points relative to stimulus onset (ms)
- Fly1–Fly6: Independent biological replicates.
- LFP (mV)
- Spike frequency (Hz)
Section: With background odor stimulation
- (S)-limonene 3x10⁻3 + background odor
- Time points relative to stimulus onset (ms)
- Fly1–Fly6
- LFP (mV)
- Spike frequency (Hz)
Section: Basal firing rate
- Basal firing (Hz)
- Fly1–Fly6
- AVE
- SEM
Section: Dose–response sensitivity
- (S)-limonene 10⁻3 to 10⁻¹: Odor concentration series used for pEC50 calculations.
- LFP pEC50
- Spike pEC50
Sheet: Statistics
Description: Statistical comparisons of basal firing and odor sensitivity with and without background odor stimulation.
Section: Wilcoxon rank-sum test (two-tailed)
Variables:
- Group 1: Condition without background odor.
- Group 2: Condition with background odor.
- P value: Statistical significance of the comparison.
Comparisons include:
- Basal firing rate (Hz)
- LFP pEC50
- Spike pEC50
Code/software
Microsoft Excel is required to view the datasets.
Access information
Other publicly accessible locations of the data:
- N/A
Data was derived from the following sources:
- ORN morphological data: Nava Gonzales et al., 2021
- Single-cell RNA sequencing datasets: McLaughlin et al 2021
- All other data was collected by the main study