Figures and data
Oviposition in Drosophila is rhythmic when registered with our semiautomated egg collection device.
A, C, G, I: Average eggs collected as a function of time in four different experiment (Canton-S, LD: A, Canton-S, DD:C; yw: G; perS: I). The average was made with all flies (Strongly rhythmic, weakly rhythmic and arrhythmic). White and dark grey bars represent periods of lights on and off respectively (LD), whereas light gray bars represent subjective days, DD, (i.e times where lights were on at rearing, but are now off). B, D, H, J: Lomb-Scargle periodograms of all genotypes, made with all flies, strongly rhythmic, weakly rhythmic and arrhythmic. Red and green horizontal lines represent significances of 0.05 and 0.01, respectively. E, K: Percentage of females with rhythmic oviposition (E: Canton-S in LD and DD, K: yw and perS). The rhythmic flies include both strongly and weakly rhythmic flies. F, L: Period of oviposition rhythms for rhythmic individual flies (F: Canton-S in LD and DD, L: YW and perS). ns: non significant, ***p<0.001 (chi-squared test for scatter plots and proportions).
Circadian rhythmicity of oviposition is dramatically reduced when the molecular clock is disrupted in all clock neurons, but not when only LNv or DN1 neurons are affected.
A, F, K, P: Schematic diagram of the neurons (painted in red) where the molecular clock has been disrupted. B, D, G, I, L, N, Q, S: Average eggs collected as a function of time. The average was made with all flies (strongly rhythmic, weakly rhythmic and arrhythmic). White and dark grey bars represent periods of lights on and off respectively (LD), whereas light gray bars represent subjective days, DD, (i.e times where lights were on at rearing, but are now off).C, E, H, J, M, O, R, T: Lomb-Scargle periodograms of all genotypes (made with all flies, rhythmic, weakly rhythmic and arrhythmic). Red and green horizontal lines represent significances of 0.05 and 0.01, respectively. B, C: +>UAS-perRNAi, n=22. D, E: Clk856-Gal4>UAS-perRNAi, n=26. G, H: +>UAS-perRNAi, n=18. I, J: PdfDicer-Gal4>UAS-perRNAi, n=34. L, M: +>UAS-perRNAi, n=38. N, O: Clk4.1-Gal4>UAS-perRNAi, n=40. Q, R: +>UAS-kir2.1, n=36. S, T: Clk4.1-Gal4>UAS-kir2.1, n=35.
Disruption of the molecular clock in Cry+ LNd neurons drastically reduces the circadian rhythmicity of oviposition.
A, H: Schematic diagram of the neurons (painted in red) where the molecular clocks have been disrupted. B, D, F, I, K, M: Average eggs collected as a function of time. The average was made with all flies (Strongly rhythmic, weakly rhythmic and arrhythmic). White and dark grey bars represent periods of lights on and off respectively (LD), whereas light gray bars represent subjective days, DD, (i.e times where lights were on at rearing, but are now off).C, E, G, J, L, N: Lomb-Scargle periodograms of all genotypes (made with all flies, rhythmic, weakly rhythmic and arrhythmic). Red and green horizontal lines represent significances of 0.05 and 0.01, respectively. B, C: +>UAS-perRNAi, n= 15. D, E: Mai179-Gal4; pdf-Gal80>+, n= 17. F, G: Mai179-Gal4; pdf-Gal80>UAS-perRNAi, n=28. I, J: +>UAS-cycDN, n=15. K, L: MB122Bsplit-Gal4>+, n=14. M, N: MB122Bsplit-Gal4>UAS-cycDN, n=30.
Disruption of the molecular clock in Cry+ LNd neurons does not alter the circadian rhythmicity of locomotor activity of mated female flies.
A: Average activity recording during 3 days in LD and 7 in DD. Light grey bars represent subjective days. B: Percent of rhythmic flies in DD. C: Periods of locomotor activity in DD of individually rhythmic flies. Mean ± SEM indicated. Each dot represents one fly. ns: non significant, *: p<0.05 (chi-squared test for the comparison of proportions, and one-way-ANOVA with post-hoc Tukey for the scatter plots.
Direct synaptic connections between circadian clock neurons and oviposition-related neurons in the hemibrain dataset.
A: Schematic diagrams showing the different neuron clusters analyzed (in color). B: Connection between Cry+ LNd and oviIN neurons. C: Connection between Cry-LNd 2and pC1b neurons. D: Network representation of the connectivity between Cry+ LNd and oviIN neurons. E: Network representation of the connectivity between Cry-LNd and pC1b neurons. Numbers give the number of synaptic contacts, which represent the strength of the connections. Only intermediate (between 3 and 9 synaptic contacts) and strong (>9 synaptic contacts) connections are considered.
Scheme of connections between circadian clock and oviposition-related neuron clusters of the same hemisphere in the hemibrain dataset.
Each circle represents a neuron cluster, comprising different numbers of neurons (some clusters comprise only one neuron). Clusters and connections involved in the connectivity between circadian clock and oviposition sets have been colored. In each connection the arrow points to the post-synaptic cluster. OviIN neurons are bidirectionally connected to every neuron of the clusters inside the square.
