Sex- and strain-dependent effects of ageing on sleep and activity patterns in Drosophila
Raw lifespan and Drosophila Activity Monitor (DAM5H) data to support the manuscript "Sex- and strain-dependent effects of ageing on sleep and activity patterns in Drosophila."
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Fly stocks and husbandry Drosophila stocks were maintained and experiments conducted at 25ºC on a 12h:12h light:dark cycle on food containing 10% (w/v) brewer’s yeast (MP Bio, lot number U1122284494-1), 5% (w/v) sucrose, 1.5% (w/v) agar, 0.3% (w/v) Nipagin, and 0.3% (v/v) propionic acid. The Canton-S (CS) stock used here was a kind gift from the lab of Colin McClure (Queen’s University Belfast). The wild-caught Dahomey (Dah) wild-type stock was collected in 1970 in Dahomey (now Benin) and has since been maintained in large populations with overlapping generations; the Dah stock used here was obtained from the lab of Linda Partridge (University College London). The w1118 stock used here was also obtained from the Partridge lab where it has been maintained in smaller population sizes. All stocks were maintained under identical conditions (multiple large population bottles) for at least three generations before being used in the present studies. Survival analysis Lifespan assays were carried out as described in detail in (Piper & Partridge, 2016). From the eggs collected from each set of parental flies, the progeny that emerged as adults within a 24-hour window were collected and allowed to mate for 48 hours, after which they were separated into single-sex vials containing standard food at a density of 15 individuals per vial. Flies were transferred to fresh vials three times per week, with deaths and censors scored during each transfer. Microsoft Excel (template available at http://piperlab.org/resources/) was used to calculate the deaths and censors in the files held here. Drosophila Activity Monitor experiments Flies were generated and maintained as for survival analysis above, with successive groups of mated female and male flies generated at 2-week intervals. When the oldest cohort of flies for a strain reached 8 weeks of age, all four cohorts (2-week, 4-week, 6-week, and 8-week) were briefly anaesthetised and transferred to single-fly tubes (Trikinetics, 65mm glass tubes) containing a small amount of standard fly food at one end and a small cotton plug at the other end. These tubes were then inserted into DAM5H Drosophila Activity Monitors (DAMs, Trikinetics) with the food side of the tube adjacent to beam number ‘1’. DAMs were then placed in an incubator at 25ºC on a 12h:12h light:dark cycle for at least 72 hours (sufficient time to gather data before any fertilised eggs produced larvae that could confound activity counts). Data were collected using the DAMSystem3 software (Trikinetics) with activity counts collected once each minute. All flies were allowed at least 24 hours to acclimatise to the DAMs; data were then extracted for 48 hours starting from the next lights-on event. During these 48 hours of data recording, the exact ages of each cohort were 13-14 days, 27-28 days, 41-42 days, and 55-56 days. Data on activity counts were analysed using the three custom R scripts available in the files here in RStudio Version 2023.03.1+446.