Fly最新文献

{"title":"Functional interplay between ribosomal protein paralogues in the eRpL22 family in Drosophila melanogaster.","authors":"Catherine M Mageeney,&nbsp;Michael G Kearse,&nbsp;Brett W Gershman,&nbsp;Caroline E Pritchard,&nbsp;Jennifer M Colquhoun,&nbsp;Vassie C Ware","doi":"10.1080/19336934.2018.1549419","DOIUrl":"https://doi.org/10.1080/19336934.2018.1549419","url":null,"abstract":"<p><p>Duplicated ribosomal protein (RP) genes in the Drosophila melanogaster eRpL22 family encode structurally-divergent and differentially-expressed rRNA-binding RPs. eRpL22 is expressed ubiquitously and eRpL22-like expression is tissue-restricted with highest levels in the adult male germline. We explored paralogue functional equivalence using the GAL4-UAS system for paralogue knockdown or overexpression and a conditional eRpL22-like knockout in a heat- shock flippase/FRT line. Ubiquitous eRpL22 knockdown with Actin-GAL4 resulted in embryonic lethality, confirming eRpL22 essentiality. eRpL22-like knockdown (60%) was insufficient to cause lethality; yet, conditional eRpL22-like knockout at one hour following egg deposition caused lethality within each developmental stage. Therefore, each paralogue is essential. Variation in timing of heat-shock-induced eRpL22-like knockout highlighted early embryogenesis as the critical period where eRpL22-like expression (not compensated for by eRpL22) is required for normal development of several organ systems, including testis development and subsequent sperm production. To determine if eRpL22-like can substitute for eRpL22, we used Actin-GAL4 for ubiquitous eRpL22 knockdown and eRpL22-like-FLAG (or FLAG-eRpL22: control) overexpression. Emergence of adults demonstrated that ubiquitous eRpL22-like-FLAG or FLAG-eRpL22 expression eliminates embryonic lethality resulting from eRpL22 depletion. Adults rescued by eRpL22-like-FLAG (but not by FLAG-eRpL22) overexpression had reduced fertility and longevity. We conclude that eRpL22 paralogue roles are not completely interchangeable and include functionally-diverse roles in development and spermatogenesis. Testis-specific paralogue knockdown revealed molecular phenotypes, including increases in eRpL22 protein and mRNA levels following eRpL22-like depletion, implicating a negative crosstalk mechanism regulating eRpL22 expression. Paralogue depletion unmasked mechanisms, yet to be defined that impact paralogue co-expression within germ cells.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 3-4","pages":"143-163"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2018.1549419","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36706857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dietary management and physical exercise can improve climbing defects and mitochondrial activity in Drosophila melanogaster parkin null mutants.","authors":"Rijan Bajracharya,&nbsp;J William O Ballard","doi":"10.1080/19336934.2018.1482139","DOIUrl":"https://doi.org/10.1080/19336934.2018.1482139","url":null,"abstract":"<p><p>Physical exercise can improve gait, balance, tremor, flexibility, grip strength and motor coordination in Parkinson's disease (PD) patients. Several lines of evidence have also shown the therapeutic potential of dietary management and supplementation in halting the progression of PD. However, there is a lack of research on the combined effects of physical activity and nutrition in the progression of PD. We test the effects exercise and dietary modification in a Drosophila model of PD. In this study, we fed Drosophila parkin mutants high protein and high carbohydrate diets without and with stearic acid (4 treatments in total). In parallel, we subjected mutants to a regimen of exercise using a purpose-built 'Power tower' exercise machine. We then measured climbing ability, aconitase activity, and basal mitochondrial ROS levels. We observed that exercising parkin mutants fed the high protein diet improved their climbing ability and increased aconitase activity. There was an additional improvement in climbing and aconitase activity in exercised parkin mutants fed the high protein diet supplemented with stearic acid. No benefits of exercise were seen in parkin mutants fed the high carbohydrate diet. Combined, these results suggest that dietary management along with physical activty has potential to improve mitochondrial biogenesis and delay the progression of PD in Drosophila parkin mutants.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 2","pages":"95-104"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2018.1482139","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36363210","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Maintenance of Rhodopsin levels in Drosophila photoreceptor and phototransduction requires Protein Kinase D.","authors":"Sudipta Ashe,&nbsp;Shweta Yadav","doi":"10.1080/19336934.2019.1565256","DOIUrl":"https://doi.org/10.1080/19336934.2019.1565256","url":null,"abstract":"<p><p>During Drosophila phototransduction, the G protein coupled receptor (GPCR) Rhodopsin (Rh1) transduces photon absorption into electrical signal via G-protein coupled activation of phospholipase C (PLC). Rh1 levels in the plasma membrane are critical for normal sensitivity to light. In this study, we report that Protein Kinase D (dPKD) regulates Rh1 homeostasis in adult photoreceptors. Although eye development and retinal structure are unaffected in the dPKD hypomorph (dPKD^H), it exhibited elevated levels of Rh1. Surprisingly, despite having elevated levels of Rh1, no defect was observed in the electrical response to light in these flies. By contrast the levels of another transmembrane protein of the photoreceptor plasma membrane, Transient receptor potential (TRP) was not altered in dPKD^H. Our results indicate that dPKD is dispensable for eye development but is required for maintaining Rh1 levels in adult photoreceptors.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 3-4","pages":"164-173"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2019.1565256","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36925649","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A model-based high throughput method for fecundity estimation in fruit fly studies.","authors":"Enoch Ng'oma,&nbsp;Elizabeth G King,&nbsp;Kevin M Middleton","doi":"10.1080/19336934.2018.1562267","DOIUrl":"https://doi.org/10.1080/19336934.2018.1562267","url":null,"abstract":"<p><p>The ability to quantify fecundity is critically important to a wide range of experimental applications, particularly in widely-used model organisms such as Drosophila melanogaster. However, the standard method of manually counting eggs is time consuming and limits the feasibility of large-scale experiments. We develop a predictive model to automate the counting of eggs from images of eggs removed from the media surface and washed onto dark filter paper. Our method uses the simple relationship between the white area in an image and the number of eggs present to create a predictive model that performs well even at high egg densities where clumping can complicate the individual identification of eggs. A cross-validation approach demonstrates our method performs well, with a correlation between predicted and manually counted values of 0.88. We show how this method can be applied to a large data set where egg densities vary widely.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 3-4","pages":"183-190"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2018.1562267","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36799350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Drosophila strain specific response to cisplatin neurotoxicity.","authors":"Christopher M Groen,&nbsp;Jewel L Podratz,&nbsp;Kevin Treb,&nbsp;Anthony J Windebank","doi":"10.1080/19336934.2019.1565257","DOIUrl":"https://doi.org/10.1080/19336934.2019.1565257","url":null,"abstract":"<p><p>Drosophila melanogaster has recently been developed as a simple, in vivo, genetic model of chemotherapy-induced peripheral neuropathy. Flies treated with the chemotherapy agent cisplatin display both a neurodegenerative phenotype and cell death in rapidly dividing follicles, mimicking the cell specific responses seen in humans. Cisplatin induces climbing deficiencies and loss of fertility in a dose dependent manner. Drosophila sensitivity to cisplatin in both cell types is affected by genetic background. We show that mutation or RNAi-based knockdown of genes known to be associated with CIPN incidence in humans affect sensitivity of flies to CIPN. Drosophila is a promising model with which to study the effect of genetics on sensitivity to CIPN.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 3-4","pages":"174-182"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2019.1565257","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36875034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The COP9 signalosome inhibits Cullin-RING E3 ubiquitin ligases independently of its deneddylase activity.","authors":"Annabelle Suisse,&nbsp;Miklós Békés,&nbsp;Tony T Huang,&nbsp;Jessica E Treisman","doi":"10.1080/19336934.2018.1429858","DOIUrl":"https://doi.org/10.1080/19336934.2018.1429858","url":null,"abstract":"<p><p>The COP9 signalosome inhibits the activity of Cullin-RING E3 ubiquitin ligases by removing Nedd8 modifications from their Cullin subunits. Neddylation renders these complexes catalytically active, but deneddylation is also necessary for them to exchange adaptor subunits and avoid auto-ubiquitination. Although deneddylation is thought to be the primary function of the COP9 signalosome, additional activities have been ascribed to some of its subunits. We recently showed that COP9 subunits protect the transcriptional repressor and tumor suppressor Capicua from two distinct modes of degradation. Deneddylation by the COP9 signalosome inactivates a Cullin 1 complex that ubiquitinates Capicua following its phosphorylation by MAP kinase in response to Epidermal Growth Factor Receptor signaling. The CSN1b subunit also stabilizes unphosphorylated Capicua to control its basal level, independently of the deneddylase function of the complex. Here we further examine the importance of deneddylation for COP9 functions <i>in vivo</i>. We use an uncleavable form of Nedd8 to show that preventing deneddylation does not reproduce the effects of loss of COP9. In contrast, in the presence of COP9, conjugation to uncleavable Nedd8 renders Cullins unable to promote the degradation of their substrates. Our results suggest that irreversible neddylation prolongs COP9 binding to and inhibition of Cullin-based ubiquitin ligases.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 2","pages":"118-126"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2018.1429858","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35755701","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sex-lethal in neurons controls female body growth in Drosophila.","authors":"Annick Sawala,&nbsp;Alex P Gould","doi":"10.1080/19336934.2018.1502535","DOIUrl":"https://doi.org/10.1080/19336934.2018.1502535","url":null,"abstract":"<p><p>Sexual size dimorphism (SSD), a sex difference in body size, is widespread throughout the animal kingdom, raising the question of how sex influences existing growth regulatory pathways to bring about SSD. In insects, somatic sexual differentiation has long been considered to be controlled strictly cell-autonomously. Here, we discuss our surprising finding that in Drosophila larvae, the sex determination gene Sex-lethal (Sxl) functions in neurons to non-autonomously specify SSD. We found that Sxl is required in specific neuronal subsets to upregulate female body growth, including in the neurosecretory insulin producing cells, even though insulin-like peptides themselves appear not to be involved. SSD regulation by neuronal Sxl is also independent of its known splicing targets, transformer and msl-2, suggesting that it involves a new molecular mechanism. Interestingly, SSD control by neuronal Sxl is selective for larval, not imaginal tissue types, and operates in addition to cell-autonomous effects of Sxl and Tra, which are present in both larval and imaginal tissues. Overall, our findings add to a small but growing number of studies reporting non-autonomous, likely hormonal, control of sex differences in Drosophila, and suggest that the principles of sexual differentiation in insects and mammals may be more similar than previously thought.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 2","pages":"133-141"},"PeriodicalIF":1.2,"publicationDate":"2018-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2018.1502535","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"36411248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Generation of genome-modified Drosophila cell lines using SwAP.","authors":"Alexandra Franz,&nbsp;Erich Brunner,&nbsp;Konrad Basler","doi":"10.1080/19336934.2017.1372068","DOIUrl":"https://doi.org/10.1080/19336934.2017.1372068","url":null,"abstract":"<p><p>The ease of generating genetically modified animals and cell lines has been markedly increased by the recent development of the versatile CRISPR/Cas9 tool. However, while the isolation of isogenic cell populations is usually straightforward for mammalian cell lines, the generation of clonal Drosophila cell lines has remained a longstanding challenge, hampered by the difficulty of getting Drosophila cells to grow at low densities. Here, we describe a highly efficient workflow to generate clonal Cas9-engineered Drosophila cell lines using a combination of cell pools, limiting dilution in conditioned medium and PCR with allele-specific primers, enabling the efficient selection of a clonal cell line with a suitable mutation profile. We validate the protocol by documenting the isolation, selection and verification of eight independently Cas9-edited armadillo mutant Drosophila cell lines. Our method provides a powerful and simple workflow that improves the utility of Drosophila cells for genetic studies with CRISPR/Cas9.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"11 4","pages":"303-311"},"PeriodicalIF":1.2,"publicationDate":"2017-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2017.1372068","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35362733","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dual role of BMP signaling in the regulation of Drosophila intestinal stem cell self-renewal.","authors":"Aiguo Tian,&nbsp;Jin Jiang","doi":"10.1080/19336934.2017.1384104","DOIUrl":"https://doi.org/10.1080/19336934.2017.1384104","url":null,"abstract":"<p><p>Many adult organs including Drosophila adult midguts rely on resident stem cells to replenish damaged cells during tissue homeostasis and regeneration. Previous studies have shown that, upon injury, intestinal stem cells (ISCs) in the midguts can increase proliferation and lineage differentiation to meet the demand for tissue repair. Our recent study has demonstrated that, in response to certain injury, midguts can expand ISC population size as an additional regenerative mechanism. We found that injury elicited by bleomycin feeding or bacterial infection increased the production of two BMP ligands (Dpp and Gbb) in enterocytes (ECs), leading to elevated BMP signaling in progenitor cells that drove an expansion of ISCs by promoting their symmetric self-renewing division. Interestingly, we also found that BMP signaling in ECs inhibits the production of Dpp and Gbb, and that this negative feedback mechanism is required to reset ISC pool size to the homeostatic state. Our findings suggest that BMP signaling exerts two opposing influences on stem cell activity depending on where it acts: BMP signaling in progenitor cells promotes ISC self-renewal while BMP signaling in ECs restricts ISC self-renewal by preventing excessive production of BMP ligands. Our results further suggest that transient expansion of ISC population in conjunction with increasing ISC proliferation provides a more effective strategy for tissue regeneration.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"11 4","pages":"297-302"},"PeriodicalIF":1.2,"publicationDate":"2017-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2017.1384104","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35441060","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of behavioral and transcriptional variation underlying CO₂ sensory neuron function and development in Drosophila.","authors":"Jia Wern Pan,&nbsp;Joi McLaughlin,&nbsp;Haining Yang,&nbsp;Charles Leo,&nbsp;Paula Rambarat,&nbsp;Sumie Okuwa,&nbsp;Anaïs Monroy-Eklund,&nbsp;Sabrina Clark,&nbsp;Corbin D Jones,&nbsp;Pelin Cayirlioglu Volkan","doi":"10.1080/19336934.2017.1344374","DOIUrl":"https://doi.org/10.1080/19336934.2017.1344374","url":null,"abstract":"<p><p>Carbon dioxide is an important environmental cue for many insects, regulating many behaviors including some that have direct human impacts. To further improve our understanding of how this system varies among closely related insect species, we examined both the behavioral response to CO₂ as well as the transcriptional profile of key developmental regulators of CO₂ sensory neurons in the olfactory system across the Drosophila genus. We found that CO₂ generally evokes repulsive behavior across most of the Drosophilids we examined, but this behavior has been lost or reduced in several lineages. Comparisons of transcriptional profiles from the developing and adult antennae for subset these species suggest that behavioral differences in some species may be due to differences in the expression of the CO₂ co-receptor Gr63a. Furthermore, these differences in Gr63a expression are correlated with changes in the expression of a few genes known to be involved in the development of the CO₂ circuit, namely dac, an important regulator of sensilla fate for sensilla that house CO₂ ORNs, and mip120, a member of the MMB/dREAM epigenetic regulatory complex that regulates CO₂ receptor expression. In contrast, most of the other known structural, molecular, and developmental components of the peripheral Drosophila CO₂ olfactory system seem to be well-conserved across all examined lineages. These findings suggest that certain components of CO₂ sensory ORN development may be more evolutionarily labile, and may contribute to differences in CO₂-evoked behavioral responses across species.</p>","PeriodicalId":12128,"journal":{"name":"Fly","volume":"11 4","pages":"239-252"},"PeriodicalIF":1.2,"publicationDate":"2017-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2017.1344374","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35114722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}