Data from: Humidity as a potential zeitgeber for circadian entrainment of insect systems
Data files
Jun 04, 2026 version files 312.97 MB
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Fig2_2026-06-04.zip
110.42 MB
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Fig3_2026-06-04.zip
5.41 MB
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Fig4_2026-06-04.zip
37.77 MB
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Fig5_2026-06-04.zip
56.54 MB
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README.md
6.61 KB
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Table_1_2026-06-04.zip
1.05 MB
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Table2_2026-06-04.zip
7.49 MB
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Table3_2026-06-04.zip
37.76 MB
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Table4_2026-06-04.zip
56.52 MB
Abstract
Humidity levels, like light and temperature, fluctuate daily yet are less predictable; however, whether humidity can entrain circadian clocks and synchronize animal behaviors with environmental variations remains unknown. Here, we investigate the circadian humidity entrainment in various insects across multiple orders. Insect species respond to humidity cycles with distinct patterns, some active during wet periods or at the arid-humid transition. When the humidity cue is removed, most species continue to show rhythmic activity associated with the previous arid-humid (AH) cycles. Fruit flies shift their activity accordingly when humidity cycles are altered and remain in the new rhythms under the following free-running conditions (FRC; constant humidity, HH). Moreover, Drosophila clock and hygrosensation mutants lack rhythmic activity during (AH) and after humidity entrainment (FRC with HH), indicating that core clock components and hygrosensors are essential for circadian entrainment. Our findings provide strong evidence that humidity is likely to serve as a potential zeitgeber for circadian entrainment in most, but not all, insect systems and will likely have broad applicability and importance across animal systems. While light and temperature act as the primary zeitgebers, understanding the mechanisms of humidity entrainment will help us better interpret the behavioral patterns of terrestrial animals, particularly those susceptible to dehydration.
All datasets are organized into subfolders based on the figures in https://doi.org/10.1038/s44323-026-00082-4 and processed according to the methods described in the manuscript. If there are questions, please contact Joshua Benoit and Shyh-Chi Chen.
For each metadata CSV file used for the analysis with the R script
Column A: raw data (txt files) from LAM25 recordings
Columns B and C: start and stop dates for the R analysis
Column D: region_D, sample locations of each LAM25 recording
Column E: status, animals' status (alive or dead) in the end of the experiment
Column F: species, experimental animal species or fly lines
Column G: replicate, 1 replication of each animal
Column H: photoperiod, light regime
Column I: sex, sex of individuals
Fig2_2026-06-04.zip
2A: Raw data of all species in AHLL from the LAM25 system and metadata CSV file used for the analysis with the R script.
2B: Activity analysis of Drosophila w1118 in AHLL (2-hour interval).
Column A: individual ID
Column B: sex, sex of experimental animals
Column C: total_beam, total activity counts of 24-hr period
Column D-I: activity counts during arid phase (2-hr interval)
Column J-O: activity counts during humid phase (2-hr interval)
2C: Activity comparison between arid and humid phases for various species in AHLL.
Column A: individual ID
Column B: mean_acti_all, mean activity counts of 24-hr period
Column C: mean_activity_arid, mean activity counts of 12-hr arid period
Column D: mean_activity_humid, mean activity counts of 12-hr humid period
2D: Spreadsheet_Rhythmicity: comparison of rhythmicity between various species in HHLL
Spreadsheet_Period: Raw data and analysis of the period of all species in HHLL
Spreadsheet_Power: Raw data and analysis of the power of all species in HHLL
2F,G: Raw data of Drosophila w1118 in AHDD and HHDD from the LAM25 system and metadata used for the analysis with the R script.
Spreadsheet_DDAH: trials with humidity cycles (AH) under constant dark (DD)
Spreadsheet_DDHH: trials in the free-running condition (HH) under constant dark (DD)
2I,J: Raw data of Drosophila Oregon in AHDD and HHDD from the LAM25 system and metadata used for the analysis with the R script.
2K,L: Raw data and analysis of the rhythmicity of Drosophila w1118 and Oregon in HHDD using ActogramJ.
Spreadsheet_Summary: summary of the results shown in Fig. 2K, L
Spreadsheet_w1118: data analysis for the rhythmicity, period, and power of w1118 in HHDD
Spreadsheet_Oregon: data analysis for the rhythmicity, period, and power of Orgeon in HHDD
Fig3_2026-06-04.zip:
Raw data of Drosophila w1118 in AH1LL, AH2LL, and HHLL from the LAM25 system and metadata used for the analysis with the R script.
Spreadsheet_LLAH1: Raw metadata for the first AH cycles (AH1)
Spreadsheet_LLAH2: Raw metadata for the second AH cycles (AH2)
Spreadsheet_LLHH: Raw metadata for the last free-running period (HH)
Fig4_2026-06-04.zip
4A: Raw data of all Drosophila clock mutants in AHLL from the LAM25 system and metadata used for the analysis with the R script.
Spreadsheet_per01: Raw metadata for per01 mutant
Spreadsheet_tim01: Raw metadata for tim01 mutant
Spreadsheet_cyc01: Raw metadata for cyc01 mutant
Spreadsheet_Clkjrk: Raw metadata for Clkjrk mutant
Spreadsheet_pdf01: Raw metadata for pdf01 mutant
4B: Data analysis of the rhythmicity of all Drosophila clock mutants in HHLL by ActogramJ.
Spreadsheet_Rhythmicity: Rhythmicity for various clock mutants from ActogramJ
Spreadsheet_Fisher Exact Test: statistics results from Fisher Exact Test
Fig5_2026-06-04.zip
5A: Raw data of all Drosophila hygrosensation mutants in AHLL from the LAM25 system and metadata used for the analysis with the R script.
Spreadsheet_Ir40a1: Raw metadata for Ir40a1 mutant
Spreadsheet_Ir25a2_Df(Ir25a): Raw metadata for Ir25a2/Df(Ir25a) mutant
Spreadsheet_Ir93a: Raw metadata for Ir93a mutant
Spreadsheet_Tmem63: Raw metadata for Tmem63 mutant
Spreadsheet_Obp59a1: Raw metadata for Obp59a1 mutant
Spreadsheet_Ir68a: Raw metadata for Ir68a mutant
Spreadsheet_Ir76b: Raw metadata for Ir76b mutant
Spreadsheet_ppk28: Raw metadata for Tmem63 mutant
Spreadsheet_Orco2: Raw metadata for Orco2 mutant
5B: Raw data and analysis of the rhythmicity of all Drosophila hygrosensation mutants in HHLL by ActogramJ.
Spreadsheet_Rhythmicity: Rhythmicity for various hygrosensation mutants from ActogramJ
Spreadsheet_Fisher Exact Test: statistics results from Fisher Exact Test
Table_1_2026-06-04.zip:
Raw data and analysis of the rhythmicity of all species in AHLL and HHLL.
Spreadsheet_period: data from ActogramJ for period analysis (Column B: species; Column C: data value)
Spreadsheet_power: data from ActogramJ for power analysis (Column B: species; Column C: data value)
Spreadsheet_Rhythmicity: Rhythmicity for various animal species from ActogramJ
Table2_2026-06-04.zip:
Raw data and analysis of the rhythmicity of w1118 and Oregon in AHDD and HHDD.
Spreadsheet_w1118_AHDD: Analysis of period, power, and rhythmicity for w1118 under AHDD condition by ActogramJ
Spreadsheet_w1118_HHDD: Analysis of period, power, and rhythmicity for w1118 under HHDD condition by ActogramJ
Spreadsheet_Oregon_AHDD: Analysis of period, power, and rhythmicity for Oregon under AHDD condition by ActogramJ
Spreadsheet_Oregon_HHDD: Analysis of period, power, and rhythmicity for Oregon under HHDD condition by ActogramJ
Table3_2026-06-04.zip
Raw data and analysis of the rhythmicity of Drosophila clock mutants in HHLL.
Spreadsheet_period: data from ActogramJ for period analysis (Column B: mutants; Column C: data value)
Spreadsheet_power: data from ActogramJ for power analysis (Column B: mutants; Column C: data value)
Spreadsheet_Rhythmicity: Rhythmicity for various mutants from ActogramJ
Spreadsheet_Fisher Exact Test: statistics results from Fisher Exact Test
Table4_2026-06-04.zip
Raw data and analysis of the rhythmicity of Drosophila hygrosensation mutants in HHLL.
Spreadsheet_period: data from ActogramJ for period analysis (Column B: mutants; Column C: data value)
Spreadsheet_power: data from ActogramJ for power analysis (Column B: mutants; Column C: data value)
Spreadsheet_Rhythmicity: Rhythmicity for various mutants from ActogramJ
Spreadsheet_Fisher Exact Test: statistics results from Fisher Exact Test
Additional information can be found on the Trikinetics website: https://trikinetics.com/Downloads/DAMSystem3%20Software%20Data%20Sheet.pdf
