Neuroscience

Circadian control of a sex-specific behaviour in Drosophila

Sabrina Riva
M Fernanda Ceriani
Sebastián Risau-Gusman author has email address
D Lorena Franco author has email address

Medical Physics Department, Bariloche Atomic Center, Comisión Nacional de Energía Atómica (CNEA) and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), San Carlos de Bariloche, Argentina
Laboratorio de Genética del Comportamiento. Fundación Instituto Leloir - IIBBA - CONICET, Buenos Aires, Argentina

https://doi.org/10.7554/eLife.103359.1

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Figures and data

Oviposition in Drosophila is rhythmic when registered with our automated egg collection device.
A, C, G, I: average eggs collected as a function of time (Canton-S in LD: A; Canton-S in DD: C; yw: G; per^S: I). 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: their respective Lomb-Scargle periodograms (Canton-S in LD (period 24.13 h ± 1.93 h): B; Canton-S in DD (period 25.21 h ± 2.17 h): D; yw (period 22.30 h ± 1.71 h): H; per^S (period 18.70 h ± 1.63 h): J). 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 per^S). F, L: Period of oviposition rhythms for individual flies (F: Canton-S in LD and DD, J: yw and per^S). ns: non significant, ***p<0.001 (chi-squared test).

Circadian rhythmicity of oviposition is abolished when the molecular clock is disrupted in all clock neurons, but not when only LNv or DN1 neurons are affected.
A, C, E, G: Schematic diagram of the neurons (painted in red) where the molecular clock has been disrupted. B, D, F, G: Periodograms of the time series of the average number of eggs laid. Red and green horizontal lines represent significances of 0.05 and 0.01, respectively. A, B: Clk856>per^RNAi (Clk856-Gal4>UAS-per^RNAi, n=26), Control (+>UAS-per^RNAi, period: 23.20 h ± 2.71 h n=22). C, D: Pdf>per^RNAi (PdfDicer-Gal4>UAS-per^RNAi, period: 20.74 h ± 1.21 h, n=34), control (+>UAS-per^RNAi, period: 24.34 h ± 1.83 h, n=18). E, F: Clk4.1>per^RNAi(Clk4.1-Gal4>UAS-per^RNAi, period: 22.30 h ± 1.68 h, n=40), control (+>UAS-per^RNAi, period: 23.20 h ± 1.69 h, n=38). G, H: Clk4.1>kir2.1 (Clk4.1-Gal4>UAS-kir2.1, period: 24.16 h ± 2.76 h, n=35), control (+>UAS-kir2.1, period: 24.16 h ± 2.09 h, n=36).

Disruption of the molecular clock in Cry+ LNd neurons drastically reduces the circadian rhythmicity of oviposition.
A, C: Schematic diagram of the neurons (painted in red) where the molecular clocks have been disrupted. B, D: Periodograms of the time series of the total number of eggs laid by females. Red and green horizontal lines represent significances of 0.05 and 0.01, respectively. A, B: MB122B>per^RNAi (MB122B-splitGal4>UAS-per^RNAi, n=21), control (+>UAS-per^RNAi, period: 22.30 h ± 1.93 h, n=25). C, D: Mai179; PdfG80>per^RNAi (Mai179-Gal4; pdf-Gal80>UAS-per^RNAi, n=19), control (+>UAS-per^RNAi, period: 24.16 h ± 2.24 h, n=14).

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, shading represents average + SEM. Light grey bars represent subjective days. B: Percent of rhythmic flies in DD. C: Periods of locomotor activity in DD of individual flies. Mean ± SEM indicated. Each dot represents one fly. Genotypes analyzed: MB122B>per^RNAi(MB122B-splitGal4>UAS-per^RNAi), control1 (+>UAS-per^RNAi), control2 (MB122B-splitGal4>+). ns: non significant, *: p<0.05 (one-way-ANOVA with post-hoc Tukey).

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.

All fly strains used in this study.

All fly strains used in this study.

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