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Correlative imaging of gene expression and metabolic activity by combining single-molecule mRNA FISH and nanometer-scale secondary ion mass spectrometry

  • Populations of genetically identical cells that share the same environment can differ markedly in their phenotypes. This phenomenon has been termed phenotypic heterogeneity. While a few molecular mechanisms that lead to heterogeneity in gene expression have been elucidated, it remains unclear how heterogeneity in gene expression is transmitted to heterogeneity in activity; especially in metabolism. Metabolic activity of single bacterial cells can be quantified by labeling the substrate with stable isotopes and by measuring label uptake with nanometer-scale secondary ion mass spectrometry (NanoSIMS). Here we combined NanoSIMS with single-molecule mRNA fluorescence in situ hybridization (smFISH) to link heterogeneity in gene expression and metabolism in nitrogen fixing bacteria. We find that gene expression and metabolic activity are decoupled in single cells. However, heterogeneity in gene expression is correlated with heterogeneity in metabolic activity on the population level. GenePopulations of genetically identical cells that share the same environment can differ markedly in their phenotypes. This phenomenon has been termed phenotypic heterogeneity. While a few molecular mechanisms that lead to heterogeneity in gene expression have been elucidated, it remains unclear how heterogeneity in gene expression is transmitted to heterogeneity in activity; especially in metabolism. Metabolic activity of single bacterial cells can be quantified by labeling the substrate with stable isotopes and by measuring label uptake with nanometer-scale secondary ion mass spectrometry (NanoSIMS). Here we combined NanoSIMS with single-molecule mRNA fluorescence in situ hybridization (smFISH) to link heterogeneity in gene expression and metabolism in nitrogen fixing bacteria. We find that gene expression and metabolic activity are decoupled in single cells. However, heterogeneity in gene expression is correlated with heterogeneity in metabolic activity on the population level. Gene expression kinetics can provide insights into the molecular mechanisms that lead to heterogeneity in metabolism.zeige mehrzeige weniger

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Autor*innen:Frank SchreiberORCiD
Koautor*innen:Martin Ackermann, S. Littmann, S. Escrig, G. Lavik, A. Meibom, M. Kuypers
Dokumenttyp:Posterpräsentation
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2016
DDC-Klassifikation:Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik
Freie Schlagwörter:NanoSIMS; Stable isotopes; mRNA-FISH
Veranstaltung:22nd International Workshop on “Single Molecule Spectroscopy and Super-resolution Microscopy in the Life Sciences”
Veranstaltungsort:Berlin, Germany
Beginndatum der Veranstaltung:14.09.2016
Enddatum der Veranstaltung:16.09.2016
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:13.10.2016
Referierte Publikation:Nein
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