A gain-of-function screen to identify genes that reduce lifespan in the adult of Drosophila melanogaster

Nakayama, Minoru; Ishibashi, Takehiko; Ishikawa, Hiroyasu; Sato, Hiroyasu; Usui, Takao; Okuda, Tomohiro; Yashiro, H.; Ishikawa, H.; Taikou, Yoshie; Minami, Asako; Kato, Kengo; Taki, Masataka; Aigaki, Toshiro; Gunji, Wataru; Ohtsu, Masaya; Murakami, Yasufumi; Tanuma, Sei Ichi; Tsuboi, Alice; Adachi, Mai; Kuroda, Junpei; Sasamura, Takeshi; Yamakawa, Tomoko; Matsuno, Kenji

doi:10.1186/1471-2156-15-46

Cited by 8 publications

(7 citation statements)

References 52 publications

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“…Information on these genes and their expression profiles is currently scarce. However, both genes were found to regulate Drosophila lifespan [ 37 , 38 ], suggesting some caution in the interpretation the longevity experiments, in which the elav -Gal4 drivers were used [ 39 – 41 ]. In addition, the CG16779 gene is predicted to encode a factor involved in regulation of Choline acetyltransferase (ChAT), which is responsible for the synthesis of the neurotransmitter acetylcholine [ 42 ].…”

Section: Discussionmentioning

confidence: 99%

Molecular and cytological analysis of widely-used Gal4 driver lines for Drosophila neurobiology

et al. 2020

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Background The Drosophila central nervous system (CNS) is a convenient model system for the study of the molecular mechanisms of conserved neurobiological processes. The manipulation of gene activity in specific cell types and subtypes of the Drosophila CNS is frequently achieved by employing the binary Gal4/UAS system. However, many Gal4 driver lines available from the Bloomington Drosophila Stock Center (BDSC) and commonly used in Drosophila neurobiology are still not well characterized. Among these are three lines with Gal4 driven by the elav promoter (BDSC #8760, #8765, and #458), one line with Gal4 driven by the repo promoter (BDSC #7415), and the 69B-Gal4 line (BDSC #1774). For most of these lines, the exact insertion sites of the transgenes and the detailed expression patterns of Gal4 are not known. This study is aimed at filling these gaps. Results We have mapped the genomic location of the Gal4-bearing P-elements carried by the BDSC lines #8760, #8765, #458, #7415, and #1774. In addition, for each of these lines, we have analyzed the Gal4-driven GFP expression pattern in the third instar larval CNS and eye-antennal imaginal discs. Localizations of the endogenous Elav and Repo proteins were used as markers of neuronal and glial cells, respectively. Conclusions We provide a mini-atlas of the spatial activity of Gal4 drivers that are widely used for the expression of UAS–target genes in the Drosophila CNS. The data will be helpful for planning experiments with these drivers and for the correct interpretation of the results.

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Section: Discussionmentioning

confidence: 99%

Molecular and cytological analysis of widely-used Gal4 driver lines for Drosophila neurobiology

et al. 2020

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“…A number of genes within Q4 show expression variation between young and old animals in our study (Additional file 15 : Table S7). This set includes CG8032 , which is also the only member of a set of 39 lifespan-reducing loci identified in a gain-of-function screen that is implicated by QTL mapped in the present study [ 22 ], and Nmdmc , overexpression of which has been shown to extend lifespan in flies [ 81 ]. Given the number of genes within Q4, and the ample evidence of multiple candidates present in the region, it is not unlikely that more than one gene in the region is responsible for the QTL we map.…”

Section: Discussionmentioning

confidence: 99%

“…In model systems, two broad strategies can be implemented to identify genes and pathways impacting lifespan and age-related phenotypes: Mutational analyses, and mapping loci contributing to variation in lifespan in natural, or semi-natural laboratory populations. Given the relative ease with which large-effect mutations can be generated and interrogated in flies, multiple studies have screened large sets of induced mutations for their effects on lifespan (e.g., [ 21 , 22 ]), and detailed mechanistic studies targeting specific genes and pathways have added considerably to our understanding of the aging process. However, such loci may be distinct from those that harbor naturally-segregating sites underlying variation in lifespan (compare Tables one, two, and three in [ 23 ]).…”

Section: Introductionmentioning

confidence: 99%

Genetic analysis of variation in lifespan using a multiparental advanced intercross Drosophila mapping population

2016

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BackgroundConsiderable natural variation for lifespan exists within human and animal populations. Genetically dissecting this variation can elucidate the pathways and genes involved in aging, and help uncover the genetic mechanisms underlying risk for age-related diseases. Studying aging in model systems is attractive due to their relatively short lifespan, and the ability to carry out programmed crosses under environmentally-controlled conditions. Here we investigate the genetic architecture of lifespan using the Drosophila Synthetic Population Resource (DSPR), a multiparental advanced intercross mapping population.ResultsWe measured lifespan in females from 805 DSPR lines, mapping five QTL (Quantitative Trait Loci) that each contribute 4–5 % to among-line lifespan variation in the DSPR. Each of these QTL co-localizes with the position of at least one QTL mapped in 13 previous studies of lifespan variation in flies. However, given that these studies implicate >90 % of the genome in the control of lifespan, this level of overlap is unsurprising. DSPR QTL intervals harbor 11–155 protein-coding genes, and we used RNAseq on samples of young and old flies to help resolve pathways affecting lifespan, and identify potentially causative loci present within mapped QTL intervals. Broad age-related patterns of expression revealed by these data recapitulate results from previous work. For example, we see an increase in antimicrobial defense gene expression with age, and a decrease in expression of genes involved in the electron transport chain. Several genes within QTL intervals are highlighted by our RNAseq data, such as Relish, a critical immune response gene, that shows increased expression with age, and UQCR-14, a gene involved in mitochondrial electron transport, that has reduced expression in older flies.ConclusionsThe five QTL we isolate collectively explain a considerable fraction of the genetic variation for female lifespan in the DSPR, and implicate modest numbers of genes. In several cases the candidate loci we highlight reside in biological pathways already implicated in the control of lifespan variation. Thus, our results provide further evidence that functional genetics tests targeting these genes will be fruitful, lead to the identification of natural sequence variants contributing to lifespan variation, and help uncover the mechanisms of aging.Electronic supplementary materialThe online version of this article (doi:10.1186/s12863-016-0419-9) contains supplementary material, which is available to authorized users.

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“…To test this hypothesis, we checked the intestinal morphology of the larvae, revealing that LN diet causes shortening of the intestines in the control flies, which was prevented by AOX expression (Figure 3A). We next tested whether tissue-restricted expression of AOX could reproduce the pupal lethality observed with the ubiquitous expression system, by the use of a GAL4 driver supporting expression principally in cells of the gut, as well as some other epithelia (38)(39)(40). However, pupal viability remained as high as in the controls (Figure 3B), indicating that the lethal phenotype is a consequence of AOX expression in one or more other tissues, and could be a systemic effect in which the digestive tract is significantly affected, perhaps as part of a compensatory physiological response.…”

Section: Aox Expression Induces Functional and Morphological Changes In The Larval Digestive Systemmentioning

confidence: 99%

Systemic impact of the expression of the mitochondrial alternative oxidase onDrosophiladevelopment

Camargo

Saari

Garcia

et al. 2021

Preprint

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Despite the beneficial effects shown when the mitochondrial alternative oxidase AOX from Ciona intestinalis (Tunicata: Ascidiacea) is xenotopically expressed in mammalian and insect models, important detrimental outcomes have also been reported, raising concerns regarding its envisioned deployment as a therapy enzyme for human mitochondrial and related diseases. Because of its non-proton pumping terminal oxidase activity, AOX can bypass the cytochrome c segment of the respiratory chain and alleviate the possible overload of electrons that occurs upon oxidative phosphorylation (OXPHOS) dysfunction, not contributing though to the proton-motive force needed for mitochondrial ATP synthesis. We have shown previously that AOX-expressing flies present a dramatic drop in pupal viability when the larvae are cultured on a low nutrient diet, indicating that AOX interferes with normal developmental metabolism. Here, we applied combined omics analyses to show that the interaction between low nutrient diet and AOX expression causes a general alteration of larval amino acid metabolism and lipid accumulation, which are associated with functional and morphological alterations of the larval digestive tract and with a drastic decrease in larval biomass accumulation. Pupae at the pre-lethality stage present a general downregulation of mitochondrial metabolism and a signature for starvation and deregulated signaling processes. This AOX-induced lethality is partially rescued when the low nutrient diet is supplemented with tryptophan and/or methionine. The developmental dependence on these amino acids, associated with elevated levels of lactate dehydrogenase, lactate, 2-hydroxyglutarate, choline-containing metabolites and breakdown products of membrane phospholipids, indicates that AOX expression promotes tissue proliferation and growth of the Drosophila larvae, but this is ultimately limited by energy dissipation via mitochondrial uncoupling. We speculate that the combination of diet and AOX expression may be used for the metabolic regulation of proliferative tissues, such as tumors.

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A gain-of-function screen to identify genes that reduce lifespan in the adult of Drosophila melanogaster

Cited by 8 publications

References 52 publications

Molecular and cytological analysis of widely-used Gal4 driver lines for Drosophila neurobiology

Molecular and cytological analysis of widely-used Gal4 driver lines for Drosophila neurobiology

Genetic analysis of variation in lifespan using a multiparental advanced intercross Drosophila mapping population

Systemic impact of the expression of the mitochondrial alternative oxidase onDrosophiladevelopment

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