Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus
Under stressful conditions some microorganisms adopt a quiescent stage characterized by a reversible non or slow proliferative condition that allows their survival. This adaptation was only recently discovered in Leishmania. We developed an in vitro model and a biosensor to track quiescence at popul...
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2019
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| author | Marlene Jara Ilse Maes Hideo Imamura Malgorzata A. Domagalska Jean‐Claude Dujardin Jorge Arévalo |
| author_browse | Hideo Imamura Ilse Maes Jean‐Claude Dujardin Jorge Arévalo Malgorzata A. Domagalska Marlene Jara |
| author_facet | Marlene Jara Ilse Maes Hideo Imamura Malgorzata A. Domagalska Jean‐Claude Dujardin Jorge Arévalo |
| author_sort | Marlene Jara |
| collection | Producción científica |
| description | Under stressful conditions some microorganisms adopt a quiescent stage characterized by a reversible non or slow proliferative condition that allows their survival. This adaptation was only recently discovered in Leishmania. We developed an in vitro model and a biosensor to track quiescence at population and single cell levels. The biosensor is a GFP reporter gene integrated within the 18S rDNA locus, which allows monitoring the expression of 18S rRNA (rGFP expression). We showed that rGFP expression decreased significantly and rapidly during the transition from extracellular promastigotes to intracellular amastigotes and that it was coupled in vitro with a decrease in replication as measured by BrdU incorporation. rGFP expression was useful to track the reversibility of quiescence in live cells and showed for the first time the heterogeneity of physiological stages among the population of amastigotes in which shallow and deep quiescent stages may coexist. We also validated the use of rGFP expression as a biosensor in animal models of latent infection. Our models and biosensor should allow further characterization of quiescence at metabolic and molecular level. |
| format | Article |
| id | oai:cris.upeu.edu.pe:20.500.15037/923 |
| institution | UPeU |
| language | English |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
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| spelling | oai:cris.upeu.edu.pe:20.500.15037/9232026-10-30T16:14:59ZTracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locusMarlene JaraIlse MaesHideo ImamuraMalgorzata A. DomagalskaJean‐Claude DujardinJorge ArévaloAmastigoteBiologyGreen fluorescent proteinPopulationCell biologyGene expressionLocus (genetics)LeishmaniaGeneReporter geneMolecular biologyGeneticsParasite hostingWorld Wide WebComputer scienceSociologyDemographyAmastigoteBiologyGreen fluorescent proteinPopulationCell biologyGene expressionLocus (genetics)LeishmaniaGeneReporter geneMolecular biologyGeneticsParasite hostingMicroorganisms, Genetically-ModifiedMicroorganisms, Genetically-ModifiedMicroorganisms, Genetically-ModifiedAnimalsAnimalsAnimalsDNA, Ribosomal geneticsDNA, Ribosomal geneticsDNA, Ribosomal geneticsDNA, Ribosomal metabolismDNA, Ribosomal metabolismDNA, Ribosomal metabolismLeishmania braziliensis cytologyLeishmania braziliensis cytologyLeishmania braziliensis cytologyLeishmania braziliensis growth & developmentLeishmania braziliensis growth & developmentLeishmania braziliensis growth & developmentLeishmania braziliensis metabolismLeishmania braziliensis metabolismLeishmania braziliensis metabolismLeishmania mexicana cytologyLeishmania mexicana cytologyLeishmania mexicana cytologyLeishmania mexicana geneticsLeishmania mexicana geneticsLeishmania mexicana geneticsLeishmania mexicana metabolismLeishmania mexicana metabolismLeishmania mexicana metabolismDNA, Protozoan geneticsDNA, Protozoan geneticsDNA, Protozoan geneticsDNA, Protozoan metabolismDNA, Protozoan metabolismDNA, Protozoan metabolismGreen Fluorescent Proteins biosynthesisGreen Fluorescent Proteins biosynthesisGreen Fluorescent Proteins biosynthesisGreen Fluorescent Proteins geneticsGreen Fluorescent Proteins geneticsGreen Fluorescent Proteins geneticsMiceMiceMiceGenetic LociGenetic LociGenetic LociHealth Sciences Medicine Public Health, Environmental and Occupational Health Research on Leishmaniasis StudiesHealth Sciences Medicine Epidemiology Trypanosoma species research and implicationsLife Sciences Biochemistry, Genetics and Molecular Biology Molecular Biology Biochemical and Molecular ResearchUnder stressful conditions some microorganisms adopt a quiescent stage characterized by a reversible non or slow proliferative condition that allows their survival. This adaptation was only recently discovered in Leishmania. We developed an in vitro model and a biosensor to track quiescence at population and single cell levels. The biosensor is a GFP reporter gene integrated within the 18S rDNA locus, which allows monitoring the expression of 18S rRNA (rGFP expression). We showed that rGFP expression decreased significantly and rapidly during the transition from extracellular promastigotes to intracellular amastigotes and that it was coupled in vitro with a decrease in replication as measured by BrdU incorporation. rGFP expression was useful to track the reversibility of quiescence in live cells and showed for the first time the heterogeneity of physiological stages among the population of amastigotes in which shallow and deep quiescent stages may coexist. We also validated the use of rGFP expression as a biosensor in animal models of latent infection. Our models and biosensor should allow further characterization of quiescence at metabolic and molecular level.Articlehttps://pubmed.ncbi.nlm.nih.gov/31831818https://www.nature.com/articles/s41598-019-55486-z.pdfhttps://doi.org/10.1038/s41598-019-55486-z10.1038/s41598-019-55486-zhttps://hdl.handle.net/20.500.15037/923W2996380320enScientific Reports2045-23222019-12-122026-06-13T09:40:34Z2026-06-13T09:40:34Z |
| spellingShingle | Amastigote Biology Green fluorescent protein Population Cell biology Gene expression Locus (genetics) Leishmania Gene Reporter gene Molecular biology Genetics Parasite hosting World Wide Web Computer science Sociology Demography Amastigote Biology Green fluorescent protein Population Cell biology Gene expression Locus (genetics) Leishmania Gene Reporter gene Molecular biology Genetics Parasite hosting Microorganisms, Genetically-Modified Microorganisms, Genetically-Modified Microorganisms, Genetically-Modified Animals Animals Animals DNA, Ribosomal genetics DNA, Ribosomal genetics DNA, Ribosomal genetics DNA, Ribosomal metabolism DNA, Ribosomal metabolism DNA, Ribosomal metabolism Leishmania braziliensis cytology Leishmania braziliensis cytology Leishmania braziliensis cytology Leishmania braziliensis growth & development Leishmania braziliensis growth & development Leishmania braziliensis growth & development Leishmania braziliensis metabolism Leishmania braziliensis metabolism Leishmania braziliensis metabolism Leishmania mexicana cytology Leishmania mexicana cytology Leishmania mexicana cytology Leishmania mexicana genetics Leishmania mexicana genetics Leishmania mexicana genetics Leishmania mexicana metabolism Leishmania mexicana metabolism Leishmania mexicana metabolism DNA, Protozoan genetics DNA, Protozoan genetics DNA, Protozoan genetics DNA, Protozoan metabolism DNA, Protozoan metabolism DNA, Protozoan metabolism Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins genetics Green Fluorescent Proteins genetics Green Fluorescent Proteins genetics Mice Mice Mice Genetic Loci Genetic Loci Genetic Loci Health Sciences Medicine Public Health, Environmental and Occupational Health Research on Leishmaniasis Studies Health Sciences Medicine Epidemiology Trypanosoma species research and implications Life Sciences Biochemistry, Genetics and Molecular Biology Molecular Biology Biochemical and Molecular Research Marlene Jara Ilse Maes Hideo Imamura Malgorzata A. Domagalska Jean‐Claude Dujardin Jorge Arévalo Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus |
| title | Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus |
| title_full | Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus |
| title_fullStr | Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus |
| title_full_unstemmed | Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus |
| title_short | Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus |
| title_sort | tracking of quiescence in leishmania by quantifying the expression of gfp in the ribosomal dna locus |
| topic | Amastigote Biology Green fluorescent protein Population Cell biology Gene expression Locus (genetics) Leishmania Gene Reporter gene Molecular biology Genetics Parasite hosting World Wide Web Computer science Sociology Demography Amastigote Biology Green fluorescent protein Population Cell biology Gene expression Locus (genetics) Leishmania Gene Reporter gene Molecular biology Genetics Parasite hosting Microorganisms, Genetically-Modified Microorganisms, Genetically-Modified Microorganisms, Genetically-Modified Animals Animals Animals DNA, Ribosomal genetics DNA, Ribosomal genetics DNA, Ribosomal genetics DNA, Ribosomal metabolism DNA, Ribosomal metabolism DNA, Ribosomal metabolism Leishmania braziliensis cytology Leishmania braziliensis cytology Leishmania braziliensis cytology Leishmania braziliensis growth & development Leishmania braziliensis growth & development Leishmania braziliensis growth & development Leishmania braziliensis metabolism Leishmania braziliensis metabolism Leishmania braziliensis metabolism Leishmania mexicana cytology Leishmania mexicana cytology Leishmania mexicana cytology Leishmania mexicana genetics Leishmania mexicana genetics Leishmania mexicana genetics Leishmania mexicana metabolism Leishmania mexicana metabolism Leishmania mexicana metabolism DNA, Protozoan genetics DNA, Protozoan genetics DNA, Protozoan genetics DNA, Protozoan metabolism DNA, Protozoan metabolism DNA, Protozoan metabolism Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins genetics Green Fluorescent Proteins genetics Green Fluorescent Proteins genetics Mice Mice Mice Genetic Loci Genetic Loci Genetic Loci Health Sciences Medicine Public Health, Environmental and Occupational Health Research on Leishmaniasis Studies Health Sciences Medicine Epidemiology Trypanosoma species research and implications Life Sciences Biochemistry, Genetics and Molecular Biology Molecular Biology Biochemical and Molecular Research |
| topic_facet | Amastigote Biology Green fluorescent protein Population Cell biology Gene expression Locus (genetics) Leishmania Gene Reporter gene Molecular biology Genetics Parasite hosting World Wide Web Computer science Sociology Demography Amastigote Biology Green fluorescent protein Population Cell biology Gene expression Locus (genetics) Leishmania Gene Reporter gene Molecular biology Genetics Parasite hosting Microorganisms, Genetically-Modified Microorganisms, Genetically-Modified Microorganisms, Genetically-Modified Animals Animals Animals DNA, Ribosomal genetics DNA, Ribosomal genetics DNA, Ribosomal genetics DNA, Ribosomal metabolism DNA, Ribosomal metabolism DNA, Ribosomal metabolism Leishmania braziliensis cytology Leishmania braziliensis cytology Leishmania braziliensis cytology Leishmania braziliensis growth & development Leishmania braziliensis growth & development Leishmania braziliensis growth & development Leishmania braziliensis metabolism Leishmania braziliensis metabolism Leishmania braziliensis metabolism Leishmania mexicana cytology Leishmania mexicana cytology Leishmania mexicana cytology Leishmania mexicana genetics Leishmania mexicana genetics Leishmania mexicana genetics Leishmania mexicana metabolism Leishmania mexicana metabolism Leishmania mexicana metabolism DNA, Protozoan genetics DNA, Protozoan genetics DNA, Protozoan genetics DNA, Protozoan metabolism DNA, Protozoan metabolism DNA, Protozoan metabolism Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins biosynthesis Green Fluorescent Proteins genetics Green Fluorescent Proteins genetics Green Fluorescent Proteins genetics Mice Mice Mice Genetic Loci Genetic Loci Genetic Loci Health Sciences Medicine Public Health, Environmental and Occupational Health Research on Leishmaniasis Studies Health Sciences Medicine Epidemiology Trypanosoma species research and implications Life Sciences Biochemistry, Genetics and Molecular Biology Molecular Biology Biochemical and Molecular Research |
| url | https://pubmed.ncbi.nlm.nih.gov/31831818 https://www.nature.com/articles/s41598-019-55486-z.pdf https://doi.org/10.1038/s41598-019-55486-z https://hdl.handle.net/20.500.15037/923 |
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