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Drosophila activity monitoring system

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The Drosophila Activity Monitoring System is a laboratory equipment designed to track and record the locomotor activity of Drosophila flies. It provides objective and automated data on the movement patterns of these model organisms.

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Drosophila Activity Monitoring System (DAMS, (33 citations) Activity Monitoring System (2 citations) Single-beam Drosophila Activity Monitoring System (2 citations)

112 protocols using drosophila activity monitoring system

1

Circadian Rhythm Analysis of Female Flies

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For activity recording, unmated female flies were collected on the first day after emergence by separating females from male flies under gaseous CO2 anaesthesia. Individual female flies were transferred into glass tubes (length 5 cm, diameter 7 mm) containing sugar‐agar medium (4% sucrose, 2% agar in water). They were kept for few days in constant light (LL) (150 μW cm−2) and then recorded in the Drosophila activity monitoring system (Trikinetics Inc., Waltham, Massachusetts) in constant darkness (DD).
Vaze K.M, & Helfrich‐Förster C. (2016). Drosophila ezoana uses an hour‐glass or highly damped circadian clock for measuring night length and inducing diapause. Physiological Entomology, 41(4), 378-389.
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2

Drosophila Behavioral Analysis using GABA and 5-HTP

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Behavioral analysis was conducted according to the method reported in a previous study [4 (link)]. Behavioral analysis was performed using the LAM system (TriKinetics, Waltham, MA, USA) when Drosophila flies ingested a sucrose agar medium containing 1% GABA, 0.1% 5-HTP, or GABA/5-HTP (1% GABA and 0.1% 5-HTP). The group (minimum 20 flies) activity, which was measured by the LAM system, provided locomotor activity levels. All experiments were repeated five times.
Changes in locomotor activity by generation were measured using a Drosophila activity monitoring system (DAM, TriKinetics), and these measurements involved housing Drosophila flies individually in glass tubes. Drosophila flies were acclimated to the tube for 24 h, and all recordings were measured with an infrared detector every minute during the experiment in constant darkness at 25 ± 1 °C. Data were analyzed using the Actogram J software, and sleep parameters were calculated by summing the number of all activities recorded daily. Subjective nighttime activity was calculated as the sum of total activity, and subjective nighttime sleep was calculated as the sum of total sleep in the light and dark phases. Sleep was defined as a period of inactivity for at least 5 min (0 interruptions per minute). The number of sleep bouts was calculated by adding the sleep episodes.
Ahn Y., Hong K.B., Kim S., Suh H.J, & Jo K. (2021). Changes in Locomotor Activity and Oxidative Stress-Related Factors after the Administration of an Amino Acid Mixture by Generation and Age. International Journal of Molecular Sciences, 22(18), 9822.
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3

Circadian Rhythms in Drosophila

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Seven-day old male flies (N = 32), were transferred to small glass tubes containing the sugar-agar food medium. Vials were located in DAMS monitors (Drosophila Activity Monitoring System, TriKinetics) and placed in an incubator (25°C). Monitors were equipped with infrared sensors, which automatically recorded activity of the flies inside their vials every 5 min. For the first 5 days, monitors were held in LD 12:12 (12 h of light and 12 h of darkness) conditions and in constant darkness (DD) for the next 6 days. Results from the second day of recording were analyzed to estimate the total activity and duration of sleep during the day and night using a Microsoft Excel plugin – BeFly (kindly donated by Dr. E. Green from the Department of Genetics, University of Leicester) (Rosato and Kyriacou, 2006 (link)) and Python 221. Sleep in flies is defined as the time for which they do not change their position for at least 5 min. The experiment was repeated three times. In LD 12:12 and DD the rhythm of locomotor activity was analyzed, and its period was measured in DD.
Doktór B., Damulewicz M, & Pyza E. (2019). Overexpression of Mitochondrial Ligases Reverses Rotenone-Induced Effects in a Drosophila Model of Parkinson’s Disease. Frontiers in Neuroscience, 13, 94.
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4

Analyzing Sleep Patterns in Drosophila after TBI

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Example 10

TBI in humans also causes sleep disorders such as hypersomnia and insomnia. Our preliminary data indicated that TBI in flies affects sleep patterns. We used a well-established assay and instrument (the Drosophila Activity Monitoring System from TriKinetics) to monitor sleep and found that injured flies had increased daytime sleep relative to uninjured flies, but nighttime sleep was not affected. We will follow-up on this finding by analyzing other sleep parameters: total sleep over 24 hours, total daytime sleep, average daytime sleep bout length, maximum daytime sleep bout length, and daytime locomotor activity per waking minute. To determine the time course of sleep pattern changes, these sleep parameters are determined over 1-week periods that begin 1, 5, 10 and 20 days after injury.

US10207011B2. Methods, systems, and devices for an invertebrate model of traumatic brain injury (2019-02-19). WISCONSIN ALUMNI RESEARCH FOUNDATION [US]. Inventors: Barry S. Ganetzky [US], David A. Wassarman [US].
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5

Drosophila Models of Polyglutamine Disorders

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The following fly lines (gmr-GAL4, UAS-GGGGCC30-EGFP), (elav-GAL4, UAS-GGGGCC3-EGFP), (elav-GAL4, UAS-GGGGCC30-EGFP), (ok371-GAL4, UAS-GGGGCC3-EGFP), (ok371-GAL4, UAS-GGGGCC3-EGFP) and (gmr-GAL4, UAS-CGG90-EGFP) have been generated in previously published work (14 (link)). All Drosophila lines were maintained, and crosses were performed in standard medium at 25°C (35 (link)). NiII(Chro)2 and CoII(Chro)2 were dissolved in DMSO (Sigma), and then added to green coloured food (Kroger), an indicator for better mixture with fly food, which was administered to the fly food. The same amount of DMSO was added to the fly food as an internal control. The progeny were collected and aged to 7 days. The screen was performed by scoring the eye phenotype, which was visualized using light microscopy. For the Drosophila activity assay, equal amounts of males and females were individually placed in Drosophila Activity Monitoring System (TriKinetics Inc, Waltham, MA, USA) testing chambers that had been capped with regular food at one end. The flies were grown on a 12-h:12-h light: dark cycle at 25°C for 4 weeks. Locomotor data were collected in the light cycle at 25°C. Locomotor activity averages of each day during the fourth week were calculated. Data were assayed in eight flies from each group at a time.
Jhan C.R., Satange R., Wang S.C., Zeng J.Y., Horng Y.C., Jin P., Neidle S, & Hou M.H. (2021). Targeting the ALS/FTD-associated A-DNA kink with anthracene-based metal complex causes DNA backbone straightening and groove contraction. Nucleic Acids Research, 49(16), 9526-9538.
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6

Drosophila Sleep Monitoring Protocol

Cited 2 times
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Flies were collected under CO2 anesthesia 1- to 2-day-old post-eclosion (to allow time for mating of the females), placed in food vials in sets of 30–50 flies each, and placed in behavioral incubator for 2 days of LD. Female flies were used for all experiments except where noted. Flies were loaded into sleep tubes and entrained for 3 days in LD before proceeding into a given experiment (see below). Behavioral assays and analysis were carried out as previously described [36 (link)]. For all experiments, flies were placed individually in glass locomotor-monitoring tubes with standard media (5% sucrose, 2% agar). Locomotor activity was monitored in 1-min bins using the Drosophila Activity Monitoring System (TriKinetics). Sleep was defined as 5 consecutive minutes of inactivity. Sleep parameters were calculated and graphed using MATLAB software [27 (link)]. In all experiments (except where noted in Figures 5 and 6) in which different genotypes are compared, data are from flies collected and run concurrently. Activity while active (Figures 1E, S2, and S4B) is a measure of the intensity of locomotor activity and is independent of sleep. It is calculated by measuring the number of beam breaks during periods during which the fly is awake. Hyperlocomotor flies would be expected to show an increase in this metric, while hypoactive flies would show a decrease.
Parisky K.M., Agosto Rivera J.L., Donelson N.C., Kotecha S, & Griffith L.C. (2016). Reorganization of Sleep by Temperature in Drosophila Requires Light, the Homeostat, and the Circadian Clock. Current biology : CB, 26(7), 882-892.
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7

Measuring Locomotor Activity in Drosophila TBI

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To measure locomotor activity, individual flies (24 biological replicates/condition) for control and TBI condition were placed in tubes containing regular fly food in a Drosophila activity monitoring system which measures the number of times a given fly crosses an infrared beam (TriKinetics Inc., Waltham, MA) (44 (link)). The activity was assessed for 2 days. Flies were subjected to 12-hr light/dark cycles with activity summarized every 30 min producing 96 timepoints of data. The number of beam breaks occurring as a result of fly movement in 30-min time-bins before the specified time-point are plotted as locomotor activity for that time-point. Flies that did not live through the recording period were not used in the calculations. Repeated measures ANOVA with Fisher's Least Significant Difference (LSD) and Bonferroni for multiple comparisons test was used to compute statistical significance (p < 0.05) between control and TBI groups using SPSS.
Shah E.J., Gurdziel K, & Ruden D.M. (2020). Drosophila Exhibit Divergent Sex-Based Responses in Transcription and Motor Function After Traumatic Brain Injury. Frontiers in Neurology, 11, 511.
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8

Competitive Behavior and Locomotion in Drosophila

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Using an aspirator, individual males were introduced to an 8-dram plastic vial. Vials were gently tapped and the time for the winner, loser, or naive male to reach to the top of the vial was recorded. Each male was tested twice, and the final score was calculated as the average of the two attempts. We also measured the fly’s general locomotion using the Drosophila Activity Monitoring system (Trikinetics) (36 (link)). Individual males were placed in 65 mm × 5 mm transparent plastic tubes with standard molasses agar media and placed in the activity monitoring system. Locomotor activity data were collected in 1-min bins using flies kept in a 12 h:12 h light:dark regimen at 25 °C and 65% humidity. Sleep data were extracted from the locomotor data as described previously (37 (link)), with sleep being defined as a period of 5 min or longer of inactivity.
Kim Y.K., Saver M., Simon J., Kent C.F., Shao L., Eddison M., Agrawal P., Texada M., Truman J.W, & Heberlein U. (2018). Repetitive aggressive encounters generate a long-lasting internal state in Drosophila melanogaster males. Proceedings of the National Academy of Sciences of the United States of America, 115(5), 1099-1104.
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9

Circadian and Stimulant-Induced Locomotion in Flies

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Unmated adult male flies (2 to 3 days after eclosion) were collected, anesthetized with CO2, and transferred to glass tubes with standard fly food. The Trikinetics Drosophila Activity Monitoring System (Waltham, Massachusetts) was used to measure locomotion. For circadian locomotion, after 24 hours of acclimation, locomotion was recorded for 24 hours by beam breaks. For the AMPH- or COC-induced locomotion, flies were pretreated for 24 hours with either VEH [dimethyl sulfoxide (DMSO)] or 100 nM CK2i in standard fly food. Flies were then fed standard fly food with AMPH (10 mM) or COC (5 mM) and either VEH or CK2i, and after 1 hour of acclimation, beam breaks were measured as above for a period of 1 hour.
Shekar A., Mabry S.J., Cheng M.H., Aguilar J.I., Patel S., Zanella D., Saleeby D.P., Zhu Y., Romanazzi T., Ulery-Reynolds P., Bahar I., Carter A.M., Matthies H.J, & Galli A. (2023). Syntaxin 1 Ser14 phosphorylation is required for nonvesicular dopamine release. Science Advances, 9(2), eadd8417.
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10

Drosophila Locomotor Activity Monitoring

Cited 2 times
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Drosophila male flies were transferred individually to activity tubes containing standard food. TriKinetics Drosophila Activity Monitoring system (Waltham, MA) was used to measure basal and AMPH-induced locomotion, as previously described [17 (link), 32 (link)].
Belovich A.N., Aguilar J.I., Mabry S.J., Cheng M.H., Zanella D., Hamilton P.J., Stanislowski D.J., Shekar A., Foster J.D., Bahar I., Matthies H.J, & Galli A. (2019). A network of phosphatidylinositol (4,5)-bisphosphate (PIP2) binding sites on the dopamine transporter regulates amphetamine behavior in Drosophila Melanogaster. Molecular psychiatry, 26(8), 4417-4430.
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