Methods for studying metabolism in Drosophila

Tennessen, Jason M.; Barry, William E.; Cox, James E.; Thummel, Carl S.

doi:10.1016/j.ymeth.2014.02.034

Cited by 390 publications

(400 citation statements)

References 73 publications

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“…The homogenate was further diluted 1:6 prior to the sugar measurements. Measurements of circulating sugars were performed using a modification of the Tennessen method (Tennessen et al 2014). For a more detailed description see File S1.…”

Section: Determination Of Circulating Sugarsmentioning

confidence: 99%

“…Glycogen measurements in the 96-well plate format were based on the conversion of glycogen to glucose by amyloglucosidase (Sigma) and on its subsequent measurement by the glucose assay (GO) kit (Sigma) as described in Tennessen et al (2014). For a more detailed description see File S1.…”

Section: Glycogen Determinationmentioning

confidence: 99%

“…Glycogen measurements in the 96 well plate format were based on the conversion of glycogen to glucose by amyloglucosidase (Sigma) and on its subsequent measurement by the glucose assay (GO) kit (Sigma) as described in Tennessen et al (TENNESSEN et al 2014). Fly homogenates were diluted with 0.05% Tween-20, in order to retain the sugar amounts within the linear range of the assay (0 to 0.16 g of glycogen or glucose per l of the analyzed homogenate).…”

Section: Glycogen Determinationmentioning

confidence: 99%

“…The homogenate was further diluted 1:6 prior to the sugar measurements. Measurements of circulating sugars were performed using a modification of the Tennessen method (TENNESSEN et al 2014). Briefly, the assay was based on conversion of trehalose to glucose by porcine trehalase (Sigma), and subsequent measurement of glucose by the glucose assay (GO) kit (Sigma).…”

Section: Determination Of Circulating Sugarsmentioning

confidence: 99%

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Energy Homeostasis Control in Drosophila Adipokinetic Hormone Mutants

et al. 2015

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Maintenance of biological functions under negative energy balance depends on mobilization of storage lipids and carbohydrates in animals. In mammals, glucagon and glucocorticoid signaling mobilizes energy reserves, whereas adipokinetic hormones (AKHs) play a homologous role in insects. Numerous studies based on AKH injections and correlative studies in a broad range of insect species established the view that AKH acts as master regulator of energy mobilization during development, reproduction, and stress. In contrast to AKH, the second peptide, which is processed from the Akh encoded prohormone [termed "adipokinetic hormone precursor-related peptide" (APRP)] is functionally orphan. APRP is discussed as ecdysiotropic hormone or as scaffold peptide during AKH prohormone processing. However, as in the case of AKH, final evidence for APRP functions requires genetic mutant analysis. Here we employed CRISPR/Cas9-mediated genome engineering to create AKH and AKH plus APRP-specific mutants in the model insect Drosophila melanogaster. Lack of APRP did not affect any of the tested steroid-dependent processes. Similarly, Drosophila AKH signaling is dispensable for ontogenesis, locomotion, oogenesis, and homeostasis of lipid or carbohydrate storage until up to the end of metamorphosis. During adulthood, however, AKH regulates body fat content and the hemolymph sugar level as well as nutritional and oxidative stress responses. Finally, we provide evidence for a negative autoregulatory loop in Akh gene regulation.KEYWORDS Drosophila; adipokinetic hormone; adipokinetic hormone precursor-related peptide; energy homeostasis; stress resistance E NERGY homeostasis requires continuous compensation for fluctuations in the energy expenditure and availability of food resources. Organisms thus build up reserves under positive energy balance and catabolize them when the balance turns negative to retain stable levels of circulating energy fuel. Insulin signaling induces the uptake of excessive circulating sugars, thus promoting reserve accumulation (reviewed, e.g., in Saltiel and Kahn 2001; Cohen 2006), whereas energy mobilization is under the control of glucagon and glucocorticoid signaling in mammals (reviewed, e.g., in Rui 2014;Charron and Vuguin 2015) and adipokinetic hormone (AKH) signaling in insects (reviewed, e.g., in Van der Horst 2003;Lorenz and Gäde 2009;Bednářová et al. 2013a). Consistent with their fundamental physiological function in energy mobilization, AKHs are found not only in insects, but are common in Protostomia, where they have been identified both in Ecdyszoa (in Arthropoda, Tardigrada, and Priapulida) and Lophotrochozoa (in Mollusca, Rotifera, and Annelida) (Gäde 2009;Hauser and Grimmelikhuijzen 2014). Nevertheless, physiological functions of AKHs have been studied mainly in Arthropoda. Similar to mammals, also insects store lipids in the form of triacylglycerides (TGs) and as carbohydrates in the form of glycogen. The main storage organ for lipid and glycogen in insects is the fat body, which can thus b...

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Section: Determination Of Circulating Sugarsmentioning

confidence: 99%

Section: Glycogen Determinationmentioning

confidence: 99%

Section: Glycogen Determinationmentioning

confidence: 99%

Section: Determination Of Circulating Sugarsmentioning

confidence: 99%

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Energy Homeostasis Control in Drosophila Adipokinetic Hormone Mutants

et al. 2015

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“…Glucose, glycogen and TAGs are parameters extensively used as measures of carbohydrate and fat metabolism [29]. Glucose, glycogen and TAG levels were measured in flies of all genotypes.…”

Section: Metabolism and Gene Expressionmentioning

confidence: 99%

Longevity and stress resistance are affected by activation of TOR/Myc in progenitor cells of Drosophila gut

et al. 2017

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Diverse physiological pathways have been shown to regulate longevity, stress resistance, fecundity and feeding rates, and metabolism in Drosophila. Here we tesed physiological traits in flies with Rheb and MycRheb overexpressed in gut progenitor cells, known as enteroblasts (EBs). We found that activation of TOR signaling by overexpression of Rheb in EBs decreases survival and stress resistance. Additionall, we showed that Myc co-expression in EBs reduces fly fecundity and feeding rate. Rheb overexpression enhanced the level of whole body glucose. Higher relative expression of the metabolic genes dilps, akh, tobi and pepck was, however, observed. The role of TOR/Myc in the regulation of genes involved in lipid metabolism and protein synthesis was established. We showed a significant role of TOR/ Myc in EBs in the regulation of the JAK/STAT, EGFR and insulin signaling pathways in Drosophila gut. These results highlight the importance of the balance between all different types of cells and confirm previous studies demonstrating that promotion of homeostasis in the intestine of Drosophila may function as a mechanism for the extension of organismal lifespan. Overall, the results demonstrate a role of TOR signaling and its downstream target Myc in EB cells in the regulation of Drosophila physiological processes.

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Characterization of a Nilaparvata lugens (Stål) brummer gene and analysis of its role in lipid metabolism

Zhou

You

et al. 2017

Arch Insect Biochem Physiol

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The brummer (bmm) genes encode the lipid storage droplet-associated triacylglycerols (TAG) lipases, which belong to the Brummer/Nutrin subfamily. These enzymes hydrolyze the ester bonds in TAG in lipid metabolism and act in insect energy homeostasis. Exposure to some agricultural chemicals leads to increased fecundity, which necessarily involves lipid metabolism, in some planthopper species. However, the biological roles of bmm in planthopper lipid storage and mobilization have not been investigated. Here, the open reading frame (ORF) of bmm (Nlbmm) was cloned and sequenced from the brown planthopper (BPH; Nilaparvata lugens). The ORF is 1014 bp encoding 338 amino acid residues. Nlbmm contained patatin domains and shared considerable evolutionary conservation with other insect bmms. Nlbmm is highly expressed in the fat body, consistent with its roles in lipid metabolism. Injection with Nlbmm double-stranded RNA (dsNlbmm) led to reduced Nlbmm mRNA accumulation, but did not influence expression of several genes related to lipid synthesis including acyl-CoA-binding protein (ACBP), acetyl-CoA carboxylase (ACC), and a lipophorin receptor (LpR). Nlbmm knockdown led to increased TAG contents in whole bodies, accumulation of total fat body lipid, and decreased hemolymph lipid content. Nlbmm knockdown did not influence the synthesis and distribution of glycerol. We infer that Nlbmm acts in TAG breakdown and fat metabolism in N. lugens.

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Methods for studying metabolism in Drosophila

Cited by 390 publications

References 73 publications

Energy Homeostasis Control in Drosophila Adipokinetic Hormone Mutants

Energy Homeostasis Control in Drosophila Adipokinetic Hormone Mutants

Longevity and stress resistance are affected by activation of TOR/Myc in progenitor cells of Drosophila gut

Characterization of a Nilaparvata lugens (Stål) brummer gene and analysis of its role in lipid metabolism

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