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Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus
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Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus
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Journal Article
Tracking of quiescence in Leishmania by quantifying the expression of GFP in the ribosomal DNA locus
2019
Overview
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.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ 13/31
/ 14/10
/ 14/19
/ 14/34
/ 14/35
/ 38
/ 38/35
/ 42/109
/ 64/60
/ Animals
/ Green Fluorescent Proteins - biosynthesis
/ Green Fluorescent Proteins - genetics
/ Humanities and Social Sciences
/ Leishmania braziliensis - cytology
/ Leishmania braziliensis - growth & development
/ Leishmania braziliensis - metabolism
/ Leishmania mexicana - cytology
/ Leishmania mexicana - genetics
/ Leishmania mexicana - metabolism
/ Mice
/ Microorganisms, Genetically-Modified
/ rRNA 18S
/ Science
