Data to the manuscript "Biological aging of two innate behaviors of Drosophila melanogaster: escape climbing versus courtship learning and memory" by Thiem J, Viskadourou M, Gaitanidis A, Stravopodis DJ, Strauß R, Duch C, Consoulas C

All measurements that were used for the figures in Thiem et al., 2024 are provided in a single excel file with a separate page for each figure. 

Figure 1A, B. Climbing performance remains stable in mid life but decreases during late life

 Oregon-R males were singly isolated in falcon vials and left undisturbed for 30 minutes. Then each fly received three Light Stimuli (LS, displacement of the fly at the bottom of the vial by tapping or flipping the vial) followed by six Moderate Stimuli (MS, 1 s vortexing) in two rounds. Finally, each fly received a combination of ~20 mixed stimuli (Strong Stimuli, SS; flipping, banking and vortexing of the vial) in a fast pace. We measured the time-to-climb a 6 cm vertical distance on the wall of the vial within 10 seconds. Failure was considered the unsuccessful effort of completing the task within 10 s. The 6 cm vertical distance of the vial was divided in 4 regions (I=bottom-1.5 cm, II=1.5-3.0 cm, III=3-4.5 cm, IV=4.5-6.0 cm). The final position (I, II, III, IV) for flies failing to complete the task within 10 seconds was marked. In the case a fly failed to initiate climbing (FAILED) or responded by jump or flight instead of climbing was marked as well. All measurements are in seconds.   

Figure 2. Courtship LTM is absent during mid and late life

A. Courtship Index of males towards Tester and Sham. Males were frustrated by mated females for eight hours and their courtship towards testers (young virgins) were measured 24 hours later. Males housed in the same devises for eight hours but without the presence of a mated female were examined for their courtship performance 24 hours later (Sham).

B. Long Term Memory was estimated as the ratio CI test/the mean of CI sham.

Figure 3. Late life pathophysiology of locomotor behavior

A, B, F. The physical status of each male was examined once a day. Healthy responses or/and deficits were letter-coded.

f= fit (normal escape climbing)

a=Jump

b=flight

c=Leg immobility

i=Moderate climbing defects

l=Severe climbing defects

h=Paradoxical behavior

n=Responsivness defect

g=Terminal stage

Courtship collapse

K=Dead

C. Age of death of experimental and control males (days).

F. Average and maximum climbing speed measurements (cm/sec).

Figure 4. Male courtship index decays in old flies, but courtship learning and (STM) remain stable.

A. Percentage of courtship of males towards trainers (young mated females) was estimated for the first 10 minutes (CI initial) and the last 10 minutes (CI final) of one hour courtship period. Then the percentage of courtship of males towards the testers (young virgin females) was estimated for the first 10 minutes (CI test). Cohorts of non-frustrated males of different age were tested for their courtship performance towards young virgin females (CI Sham) within the first 10 minutes.

B. Learning Intex is the ratio: CI final/CI initial.

C. The 5min STM is the ratio CI test/the mean of CI sham.

Figure 5. Speed decays gradually during aging whereas courtship performances collapses in old flies, but a weak-to-moderate correlation is found between speed and STM

A. Climbing speed (cm/sec) of >70 days old males.

B. Courtship Index of the same males towards trainers and testers.

C. STM Index of the same males

D. Courtship collapse. Courtship Index towards trainers and testers of the same males at the day that courtship collapses as well as the previous two days.

E. Climbing speed measurements (cm/sec) of the same males at the day that courtship collapses as well as the previous two days.

F. Correlation between climbing speed and courtship index (vs. Trainer and Tester) and STM.

Figure S1 Latencies to different courtship rituals

Measurements (seconds) of courtship rituals (courtship/orientation, singing, licking attempting copulation) of the same flies as in Figure 4 in 4, 6, 8, 10 weeks of age.

Figure S2 Latencies to courtship rituals in flies older than 10 weeks

Measurements (seconds) of courtship rituals (courtship/orientation, singing, licking attempting copulation) of those flies (Figure 5) that reached >70 days of age.

The original fly videos are available upon request