IF 1.2 4区生物学

Fly Pub Date : 2021-12-01 Epub Date: 2020-12-08 DOI: 10.1080/19336934.2020.1837592

Ruijian Guo, Anna-Lena Henke, Klaus Reinhardt

引用次数: 1

IF 1.2 4区生物学

Fly Pub Date : 2021-12-01 DOI: 10.1080/19336934.2021.1943285

Howy Jacobs

引用次数: 0

A pipeline for precise and efficient genome editing by sgRNA-Cas9 RNPs in Drosophila. 利用 sgRNA-Cas9 RNPs 对果蝇进行精确高效基因组编辑的管道。

IF 1.2 4区生物学

Fly Pub Date : 2020-03-01 Epub Date: 2020-10-21 DOI: 10.1080/19336934.2020.1832416

Kevin G Nyberg, Joseph Q Nguyen, Yong-Jae Kwon, Shelby Blythe, Greg J Beitel, Richard Carthew

{"title":"A pipeline for precise and efficient genome editing by sgRNA-Cas9 RNPs in Drosophila.","authors":"Kevin G Nyberg, Joseph Q Nguyen, Yong-Jae Kwon, Shelby Blythe, Greg J Beitel, Richard Carthew","doi":"10.1080/19336934.2020.1832416","DOIUrl":"10.1080/19336934.2020.1832416","url":null,"abstract":"Genome editing via homology-directed repair (HDR) has made possible precise and deliberate modifications to gene sequences. CRISPR/Cas9-mediated HDR is the simplest means to carry this out. However, technical challenges remain to improve efficiency and broaden applicability to any genetic background of Drosophila melanogaster as well as to other Drosophila species. To address these issues, we developed a two-stage marker-assisted strategy in which embryos are injected with RNPs and pre-screened using T7EI. Using sgRNA in complex with recombinant Cas9 protein, we assayed each sgRNA for genome-cutting efficiency. We then conducted HDR using sgRNAs that efficiently cut target genes and the application of a transformation marker that generates RNAi against eyes absent. This allows for screening based on eye morphology rather than colour. These new tools can be used to make a single change or a series of allelic substitutions in a region of interest, or to create additional genetic tools such as balancer chromosomes.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"14 1-4","pages":"34-48"},"PeriodicalIF":1.2,"publicationDate":"2020-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2020.1832416","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9278192","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}

引用次数: 0

Mitochondrial dysfunction generates a growth-restraining signal linked to pyruvate in Drosophila larvae 果蝇幼虫线粒体功能障碍产生与丙酮酸相关的生长抑制信号

IF 1.2 4区生物学

Fly Pub Date : 2019-09-17 DOI: 10.1080/19336934.2019.1662266

Jack George, Tea Tuomela, E. Kemppainen, A. Nurminen, Samuel T. Braun, Cagri Yalgin, H. Jacobs

{"title":"Mitochondrial dysfunction generates a growth-restraining signal linked to pyruvate in Drosophila larvae","authors":"Jack George, Tea Tuomela, E. Kemppainen, A. Nurminen, Samuel T. Braun, Cagri Yalgin, H. Jacobs","doi":"10.1080/19336934.2019.1662266","DOIUrl":"https://doi.org/10.1080/19336934.2019.1662266","url":null,"abstract":"ABSTRACT The Drosophila bang-sensitive mutant tko25t, manifesting a global deficiency in oxidative phosphorylation due to a mitochondrial protein synthesis defect, exhibits a pronounced delay in larval development. We previously identified a number of metabolic abnormalities in tko25t larvae, including elevated pyruvate and lactate, and found the larval gut to be a crucial tissue for the regulation of larval growth in the mutant. Here we established that expression of wild-type tko in any of several other tissues of tko25t also partially alleviates developmental delay. The effects appeared to be additive, whilst knockdown of tko in a variety of specific tissues phenocopied tko25t, producing developmental delay and bang-sensitivity. These findings imply the existence of a systemic signal regulating growth in response to mitochondrial dysfunction. Drugs and RNAi-targeted on pyruvate metabolism interacted with tko25t in ways that implicated pyruvate or one of its metabolic derivatives in playing a central role in generating such a signal. RNA-seq revealed that dietary pyruvate-induced changes in transcript representation were mostly non-coherent with those produced by tko25t or high-sugar, consistent with the idea that growth regulation operates primarily at the translational and/or metabolic level.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"13 1","pages":"12 - 28"},"PeriodicalIF":1.2,"publicationDate":"2019-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2019.1662266","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48725842","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 3

A standardized nomenclature and atlas of the male terminalia of Drosophila melanogaster 黑腹果蝇雄性绝症的标准命名法和图谱

IF 1.2 4区生物学

Fly Pub Date : 2019-06-10 DOI: 10.1080/19336934.2019.1653733

Gavin R. Rice, J. David, Y. Kamimura, John P. Masly, A. McGregor, Olga Nagy, S. Noselli, M. D. Nunes, P. O'Grady, E. Sánchez-Herrero, M. Siegal, M. Toda, Mark Rebeiz, V. Courtier-Orgogozo, A. Yassin

{"title":"A standardized nomenclature and atlas of the male terminalia of Drosophila melanogaster","authors":"Gavin R. Rice, J. David, Y. Kamimura, John P. Masly, A. McGregor, Olga Nagy, S. Noselli, M. D. Nunes, P. O'Grady, E. Sánchez-Herrero, M. Siegal, M. Toda, Mark Rebeiz, V. Courtier-Orgogozo, A. Yassin","doi":"10.1080/19336934.2019.1653733","DOIUrl":"https://doi.org/10.1080/19336934.2019.1653733","url":null,"abstract":"ABSTRACT Animal terminalia represent some of the most diverse and rapidly evolving structures in the animal kingdom, and for this reason have been a mainstay in the taxonomic description of species. The terminalia of Drosophila melanogaster, with its wide range of experimental tools, have recently become the focus of increased interest in the fields of development, evolution, and behavior. However, studies from different disciplines have often used discrepant terminologies for the same anatomical structures. Consequently, the terminology of genital parts has become a barrier to integrating results from different fields, rendering it difficult to determine what parts are being referenced. We formed a consortium of researchers studying the genitalia of D. melanogaster to help establish a set of naming conventions. Here, we present a detailed visual anatomy of male genital parts, including a list of synonymous terms, and suggest practices to avoid confusion when referring to anatomical parts in future studies. The goal of this effort is to facilitate interdisciplinary communication and help newcomers orient themselves within the exciting field of Drosophila genitalia.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"13 1","pages":"51 - 64"},"PeriodicalIF":1.2,"publicationDate":"2019-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2019.1653733","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48298400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 27

A modular toolset of phiC31-based fluorescent protein tagging vectors for Drosophila. 基于phic31的果蝇荧光蛋白标记载体的模块化工具集。

IF 1.2 4区生物学

Fly Pub Date : 2019-03-01 Epub Date: 2019-03-28 DOI: 10.1080/19336934.2019.1595999

Jun Luo, Pingping Shen, Jiong Chen

{"title":"A modular toolset of phiC31-based fluorescent protein tagging vectors for Drosophila.","authors":"Jun Luo, Pingping Shen, Jiong Chen","doi":"10.1080/19336934.2019.1595999","DOIUrl":"https://doi.org/10.1080/19336934.2019.1595999","url":null,"abstract":"The Drosophila transgenic technology and fluorescent protein fusions are powerful tools to analyze protein expression patterns, subcellular localization and protein dynamics. Recently, the Drosophila transgenic technology has been improved by the highly efficient phiC31 site-specific integration system. Many new and improved fluorescent proteins with desirable advantages have been developed. However, the phiC31 system and the newly developed fluorescent proteins have not been systematically applied in Drosophila transgenic vectors. Here, we have constructed a modular toolset of C-terminal fluorescent protein fusion vectors based on phiC31 site-specific integration system for the generation of transgenic Drosophila lines. These cloning vectors contain a variety of fluorescent tags, including blue, cyan, green or red fluorescent proteins, photoactivatable or photoswitchable fluorescent proteins, fluorescent timers, photosensitizers and bimolecular fluorescence complementation tags. These vectors provide a range of transcriptional regulation options including UAST, UASP, UASC, LexAop, QUAS, Ubi, αTub67C and αTub84B promoters, and two screening marker options including white and vermilion gene. The vectors have been tested in vivo and can produce fluorescent chimeric proteins that are functional.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"13 1-4","pages":"29-41"},"PeriodicalIF":1.2,"publicationDate":"2019-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2019.1595999","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"37067893","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}

引用次数: 3

Bivalency in Drosophila embryos is associated with strong inducibility of Polycomb target genes. 果蝇胚胎的二价性与Polycomb靶基因的强诱导性有关。

IF 1.2 4区生物学

Fly Pub Date : 2019-03-01 Epub Date: 2019-05-25 DOI: 10.1080/19336934.2019.1619438

Arslan Akmammedov, Marco Geigges, Renato Paro

引用次数: 7

Extending julius seizure, a bang-sensitive gene, as a model for studying epileptogenesis: Cold shock, and a new insertional mutation. 延长julius癫痫，一个爆炸敏感基因，作为研究癫痫发生的模型:冷休克，和一个新的插入突变。

IF 1.2 4区生物学

Fly Pub Date : 2018-01-02 Epub Date: 2017-12-08 DOI: 10.1080/19336934.2017.1402993

Derek Dean, Hannah Weinstein, Seema Amin, Breelyn Karno, Emma McAvoy, Ronald Hoy, Andrew Recknagel, Casey Jarvis, David Deitcher

{"title":"Extending julius seizure, a bang-sensitive gene, as a model for studying epileptogenesis: Cold shock, and a new insertional mutation.","authors":"Derek Dean, Hannah Weinstein, Seema Amin, Breelyn Karno, Emma McAvoy, Ronald Hoy, Andrew Recknagel, Casey Jarvis, David Deitcher","doi":"10.1080/19336934.2017.1402993","DOIUrl":"https://doi.org/10.1080/19336934.2017.1402993","url":null,"abstract":"The bang-sensitive (BS) mutants of Drosophila are an important model for studying epilepsy. We recently identified a novel BS locus, julius seizure (jus), encoding a protein containing two transmembrane domains and an extracellular cysteine-rich loop. We also determined that jussda iso7.8, a previously identified BS mutation, is an allele of jus by recombination, deficiency mapping, complementation testing, and genetic rescue. RNAi knockdown revealed that jus expression is important in cholinergic neurons and that the critical stage of jus expression is the mid-pupa. Finally, we found that a functional, GFP-tagged genomic construct of jus is expressed mostly in axons of the neck connectives and of the thoracic abdominal ganglia. In this Extra View article, we show that a MiMiC GFP-tagged Jus is localized to the same nervous system regions as the GFP-tagged genomic construct, but its expression is mostly confined to cell bodies and it causes bang-sensitivity. The MiMiC GFP-tag lies in the extracellular loop while the genomic construct is tagged at the C-terminus. This suggests that the alternate position of the GFP tag may disrupt Jus protein function by altering its subcellular localization and/or stability. We also show that a small subset of jus-expressing neurons are responsible for the BS phenotype. Finally, extending the utility of the BS seizure model, we show that jus mutants exhibit cold-sensitive paralysis and are partially sensitive to strobe-induced seizures.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 1","pages":"55-61"},"PeriodicalIF":1.2,"publicationDate":"2018-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2017.1402993","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35243197","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}

引用次数: 4

Keeping it tight: The relationship between bacterial dysbiosis, septate junctions, and the intestinal barrier in Drosophila. 保持严密：果蝇体内细菌失调、隔膜连接和肠道屏障之间的关系

IF 1.2 4区生物学

Fly Pub Date : 2018-01-02 Epub Date: 2018-03-06 DOI: 10.1080/19336934.2018.1441651

Martin Resnik-Docampo, Vivien Sauer, Joseph M Schinaman, Rebecca I Clark, David W Walker, D Leanne Jones

{"title":"Keeping it tight: The relationship between bacterial dysbiosis, septate junctions, and the intestinal barrier in Drosophila.","authors":"Martin Resnik-Docampo, Vivien Sauer, Joseph M Schinaman, Rebecca I Clark, David W Walker, D Leanne Jones","doi":"10.1080/19336934.2018.1441651","DOIUrl":"10.1080/19336934.2018.1441651","url":null,"abstract":"Maladaptive changes in the intestinal flora, typically referred to as bacterial dysbiosis, have been linked to intestinal aging phenotypes, including an increase in intestinal stem cell (ISC) proliferation, activation of inflammatory pathways, and increased intestinal permeability1,2. However, the causal relationships between these phenotypes are only beginning to be unravelled. We recently characterized the age-related changes that occur to septate junctions (SJ) between adjacent, absorptive enterocytes (EC) in the fly intestine. Changes could be observed in the overall level of SJ proteins, as well as the localization of a subset of SJ proteins. Such age-related changes were particularly noticeable at tricellular junctions (TCJ)3. Acute loss of the Drosophila TCJ protein Gliotactin (Gli) in ECs led to rapid activation of stress signalling in stem cells and an increase in ISC proliferation, even under axenic conditions; a gradual disruption of the intestinal barrier was also observed. The uncoupling of changes in bacteria from alterations in ISC behaviour and loss of barrier integrity has allowed us to begin to explore the interrelationship of these intestinal aging phenotypes in more detail and has shed light on the importance of the proteins that contribute to maintenance of the intestinal barrier.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 1","pages":"34-40"},"PeriodicalIF":1.2,"publicationDate":"2018-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5927685/pdf/kfly-12-01-1441651.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35841932","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}

引用次数: 0

FijiWingsPolarity: An open source toolkit for semi-automated detection of cell polarity. FijiWingsPolarity:用于半自动化检测细胞极性的开源工具包。

IF 1.2 4区生物学

Fly Pub Date : 2018-01-02 Epub Date: 2017-12-22 DOI: 10.1080/19336934.2017.1409927

Leonard L Dobens, Anna Shipman, Jeffrey D Axelrod

{"title":"FijiWingsPolarity: An open source toolkit for semi-automated detection of cell polarity.","authors":"Leonard L Dobens, Anna Shipman, Jeffrey D Axelrod","doi":"10.1080/19336934.2017.1409927","DOIUrl":"https://doi.org/10.1080/19336934.2017.1409927","url":null,"abstract":"Epithelial cells are defined by apical-basal and planar cell polarity (PCP) signaling, the latter of which establishes an orthogonal plane of polarity in the epithelial sheet. PCP signaling is required for normal cell migration, differentiation, stem cell generation and tissue repair, and defects in PCP have been associated with developmental abnormalities, neuropathologies and cancers. While the molecular mechanism of PCP is incompletely understood, the deepest insights have come from Drosophila, where PCP is manifest in hairs and bristles across the adult cuticle and organization of the ommatidia in the eye. Fly wing cells are marked by actin-rich trichome structures produced at the distal edge of each cell in the developing wing epithelium and in a mature wing the trichomes orient collectively in the distal direction. Genetic screens have identified key PCP signaling pathway components that disrupt trichome orientation, which has been measured manually in a tedious and error prone process. Here we describe a set of image processing and pattern-recognition macros that can quantify trichome arrangements in micrographs and mark these directly by color, arrow or colored arrow to indicate trichome location, length and orientation. Nearest neighbor calculations are made to exploit local differences in orientation to better and more reliably detect and highlight local defects in trichome polarity. We demonstrate the use of these tools on trichomes in adult wing preps and on actin-rich developing trichomes in pupal wing epithelia stained with phalloidin. FijiWingsPolarity is freely available and will be of interest to a broad community of fly geneticists studying the effect of gene function on PCP.","PeriodicalId":12128,"journal":{"name":"Fly","volume":"12 1","pages":"23-33"},"PeriodicalIF":1.2,"publicationDate":"2018-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1080/19336934.2017.1409927","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"35297211","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}

引用次数: 3

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