analyzing the effect of reserpine on the sleep behavior of drosophila melanogaster using scamp2020 data package in matlab

Advancing Sleep Disorder Treatments: A &lt;em&gt;Drosophila&lt;/em&gt;-Based Screening Approach

Sleep, one of the essential behaviors necessary for human survival, is characterized by two main states: rapid eye movement (REM) sleep and non-rapid eye movement (NREM) sleep1. NREM sleep comprises three stages: N1 (the transition between wakefulness and sleep), N2 (light sleep), and N3 (deep sleep, slow wave sleep), representing the progression from wakefulness to deep sleep1. Sleep plays a crucial role in both physical and mental health2. However, aging reduces total sleep duration, sleep efficiency, slow-wave sleep percentage, and REM sleep percentage in adults3. Older individuals tend to spend more time in light sleep compared to slow-wave sleep, making them more sensitive to nocturnal awakenings. As the number of awakenings increases, average sleep time decreases, resulting in a fragmented sleep pattern in the elderly, which may be associated with excessive excitation of Hcrt neurons in mice4. Additionally, age-related declines in circadian mechanisms contribute to an earlier shift in sleep duration5,6. In combination with physical illness, psychological stress, environmental factors, and medication use, these factors make older adults more susceptible to sleep disorders, such as insomnia, REM sleep behavior disorder, narcolepsy, periodic leg movements, restless legs syndrome, and sleep-disordered breathing7,8.
Epidemiological studies have shown that sleep disorders are closely linked to chronic diseases in the elderly9, including depression10, cardiovascular disease11, and dementia12. Addressing sleep disorders plays a crucial role in improving and treating chronic diseases and enhancing the quality of life for older adults. Currently, patients primarily rely on drugs such as benzodiazepines, non-benzodiazepines, and melatonin receptor agonists to enhance sleep quality13. However, benzodiazepines can lead to downregulation of receptors and dependence after long-term use, causing severe withdrawal symptoms upon discontinuation14,15. Non-benzodiazepine drugs also carry risks, including dementia16, fractures17, and cancer18. The commonly used melatonin receptor agonist, ramelteon, reduces sleep latency but does not increase sleep duration and has hepatic function-related concerns due to extensive first-pass elimination19. Agomelatine, a melatonin receptor agonist and serotonin receptor antagonist, improves depression-related insomnia but also poses a risk of liver damage20. Consequently, there is an urgent need for safer drugs to treat or alleviate sleep disorders. However, current drug screening strategies, based on molecular and cellular experiments combined with automated systems and computer analysis, are expensive and time-consuming21. Structure-based drug design strategies, relying on receptor structure and properties, require a clear understanding of receptor three-dimensional structure and lack predictive capabilities for drug effects22.
In 2000, based on the sleep criteria proposed by Campbell and Tobler in 198423, researchers established simple animal models to study sleep24, including Drosophila melanogaster, which exhibited sleep-like states25,26. Despite anatomical differences between Drosophila and humans, many neurochemical components and signaling pathways regulating sleep in Drosophila are conserved in mammalian sleep, facilitating the study of human neurological diseases27,28. Drosophila is also extensively used in circadian rhythm studies, despite differences in core oscillators between flies and mammals29,30,31. Therefore, Drosophila serves as a valuable model organism for studying sleep behavior and conducting sleep-related drug screening.
This study proposes a cost-effective and simple phenotype-based approach for screening small-molecule drugs to treat sleep disorders using aged flies. Sleep regulation in Drosophila is highly conserved25, and the decline in sleep observed with age may be reversible through drug administration. Thus, this sleep phenotype-based screening method can intuitively reflect drug efficacy. We feed the flies with a mixture of the drug under investigation and food, monitor and record sleep behavior using the Drosophila Activity Monitor (DAM)32, and analyze the acquired data using the open-source SCAMP2020 data package in MATLAB (Figure1). Statistical analysis is performed using statistics and graphing software (see Table of Materials). As an example, we demonstrate the effectiveness of this protocol by presenting experimental data on Reserpine, a small-molecule inhibitor of the vesicular monoamine transporter reported to increase sleep33. This protocol provides a valuable approach to identify drugs for treating age-related sleep problems.

Author Spotlight: Overcoming Challenges in Drosophila Sleep Measurement Using DAM System 

High-Throughput Small Molecule Drug Screening For Age-Related Sleep Disorders Using Drosophila melanogaster

Watch this Scientific Journal Video about Analyzing the Effect of Reserpine on the Sleep Behavior of Drosophila melanogaster Using SCAMP2020 Data Package in MATLAB at JoVE.com

Analyzing the Effect of Reserpine on the Sleep Behavior of Drosophila melanogaster Using SCAMP2020 Data Package in MATLAB  

Analyzing the Effect of Reserpine on the Sleep Behavior of Drosophila melanogaster Using SCAMP2020 Data Package in MATLAB

To begin, monitor the sleep behaviors of flies using an infrared monitoring device and import the collected data in TXT format into the DAM file scan 107 software for scanning. Set the start time of the segmentation data to either 0800 or 0801 on the third morning after initiating the monitors to obtain one or 30 minute bin length data, respectively. Set the termination time to 8:00 AM on the third day post the set starting time.Adjust the bin length option to one minute and alter the output file type to channel files. Rename the file and generate the output. Now launch the SCAMP 2020 software package in MATLAB.Double click on Vecsey Sleep and Circadian Analysis MATLAB program. Added it sub-folder titled Vecsey SCAMP Scripts into the path. Locate the SCAMP.m file in that folder and run it. In the subsequent popup window sequentially select the folder named one minute and 30 minute. Then select a monitor and click on load individual C plots to preview.Inspect the image that appears and deselect the corresponding channel of any dead flies. Rename each channel in every monitor based on the corresponding drug to be tested. Select all monitors and click Analyze selected data for the analysis.Select the default option and then click on analyze for chosen bin feature. Check the export data box. Click on graph 30 minute data types for all days for selected groups, and export all data to generate the output of the results.Select the S30 file from the CSV file and locate the corresponding mean value and standard error data for each monitor. Save this data in Excel to make some adjustments before posting it into GraphPad Prism to formulate a sleep status diagram. Afterwards, locate the file named S-T-D-U-R and compute the average values of daytime, nighttime, and total sleep for each fly over three days.Paste data into GraphPad Prism to draw graphs. The sleep patterns of Reserpine and Dimethyl Sulfoxide on female and male flies over three consecutive days are shown. Quantitative analysis of the sleep behavior demonstrates a significant increase in sleep time in aged females fed Reserpine.The experiment on male flies also demonstrates a positive correlation between Reserpine concentration and the promotion of sleep.

analyzing-effect-reserpine-on-sleep-behavior-drosophila-melanogaster

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JoVE Journal

Behavior

257 次观看.  Southeast University. 首先，使用红外监测装置监测苍蝇的睡眠行为，并将收集的数据以TXT格式导入DAM文件扫描软件107进行扫描。在启动监视器后，将分段数据的开始时间设置为 0800 或 0801，以分别获得 1 分钟或 30 分钟的条柱长度数据。将终止时间设置为设置开始时间后第三天的上午 8：00。将素材箱长度选项调整为一分钟，并将输出文件类型更改为通道文件。重命名文件并生成输出。现在在...