Project description:Drosophila melanogaster adult flies fed on food containing 16 mg/ml of pentylenetetrazole (PTZ) in the food show a hyperkinetic behavior within 24 hours. Half of that concentration, i.e., 8 mg/ml, of PTZ, if fed for seven days, though doesn’t cause seizure-like behavior, results in a decreased climbing speed in flies. This change in locomotor behavior is progressive and becomes significant only on seventh day of the treatment. We also examined flies’ locomotor behavior secondary to PTZ withdrawal. Interestingly, an increased climbing speed was found to develop seven days after withdrawal. Importantly, antiepileptic drugs showed effectiveness in the above fly model. Earlier, we submitted in GEO time series of fly head microarray gene expression profiling during chronic PTZ and PTZ withdrawal phase. We also submitted previously expression profiles associated with antiepileptic drug treatment. Here, we have undertaken a different line of work. Having developed a well characterized acquired model of behavioral and gene expression plasticity, we found an opportunity here to investigate if drug exposure to adult males could cause transgenerational effect. To probe this, we carried out a systematic study at both behavioral and microarray gene expression levels. In the latter, we asked the question that do F0 testis, F1 males’ head, F1 females’ head, F1 testis, F2 males’ head and F2 females’ head show gene expression changes if F0 male parents had a history of PTZ exposure? A total of 28 microarray slides were used in this study.
Project description:Transgenerational epigenetic inheritance is a subject of immense current interest. In a newly developed Drosophila model in the laboratory, genetic ablation of insulin-producing cells (IPCs) was found to affect whole -body triglyceride levels not only in the ablated flies but also in their male-line derived, non-ablated future generations. To further characterize this genetic-factor-induced transgenerational inheritance model, we have now performed whole body microarray gene expression profiling of adult males and females with genetically ablated IPCs, and of three consecutive, paternally derived non-ablated generations of adult males and females originating from ablated males. Interestingly, like altered levels of triglycerides, transcriptomic alterations are found not only in the ablated flies but also in their male-line-derived, non-ablated future generations.
Project description:RNA was extracted from adult male and adult female Drosophila melanogaster with reversed sex-chromosome parent-of-origin (e.g. maternal-X/paternal-Y vs. paternal-X/maternal-Y) Parent-of-origin effects were assayed in X/Y males, XY/Y males, and XY/X females. Direct comparisons were made between individuals with the same karyotype (e.g. X/Y males or XY/Y males) incorporating dye-swaps.
Project description:<p>Chronic sleep loss profoundly impacts metabolic health and shortens lifespan, but studies of the mechanisms involved have focused largely on acute sleep deprivation. To identify metabolic consequences of chronically reduced sleep, we conducted unbiased metabolomics on heads of three adult Drosophila short-sleeping mutants with very different mechanisms of sleep loss: fumin (fmn), redeye (rye), and sleepless (sss). Common features included elevated ornithine and polyamines, with lipid, acyl-carnitine, and TCA cycle changes suggesting mitochondrial dysfunction. Studies of excretion demonstrate inefficient nitrogen elimination in adult sleep mutants, likely contributing to their polyamine accumulation. Increasing levels of polyamines, particularly putrescine, promote sleep in control flies but poison sleep mutants. This parallels the broadly enhanced toxicity of high dietary nitrogen load from protein in chronically sleep-restricted Drosophila, including both sleep mutants and flies with hyper-activated wake-promoting neurons. Together, our results implicate nitrogen stress as a novel mechanism linking chronic sleep loss to adverse health outcomes-and perhaps for linking food and sleep homeostasis at the cellular level in healthy organisms.</p>
