Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell.
Chlamydia trachomatis is an obligate intracellular bacterium that alternates between two metabolically different developmental forms. We performed fluorescence lifetime imaging (FLIM) of the metabolic coenzymes, reduced nicotinamide adenine dinucleotides [NAD(P)H], by two-photon microscopy for separ...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2011-07-01
|
Series: | PLoS Pathogens |
Online Access: | http://europepmc.org/articles/PMC3136453?pdf=render |
id |
doaj-efccb16bf3a1447fa0ec1a8c37683cb3 |
---|---|
record_format |
Article |
spelling |
doaj-efccb16bf3a1447fa0ec1a8c37683cb32020-11-25T01:13:39ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742011-07-0177e100210810.1371/journal.ppat.1002108Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell.Márta SzaszákPhilipp StevenKensuke ShimaRegina Orzekowsky-SchröderGereon HüttmannInke R KönigWerner SolbachJan RuppChlamydia trachomatis is an obligate intracellular bacterium that alternates between two metabolically different developmental forms. We performed fluorescence lifetime imaging (FLIM) of the metabolic coenzymes, reduced nicotinamide adenine dinucleotides [NAD(P)H], by two-photon microscopy for separate analysis of host and pathogen metabolism during intracellular chlamydial infections. NAD(P)H autofluorescence was detected inside the chlamydial inclusion and showed enhanced signal intensity on the inclusion membrane as demonstrated by the co-localization with the 14-3-3β host cell protein. An increase of the fluorescence lifetime of protein-bound NAD(P)H [τ₂-NAD(P)H] inside the chlamydial inclusion strongly correlated with enhanced metabolic activity of chlamydial reticulate bodies during the mid-phase of infection. Inhibition of host cell metabolism that resulted in aberrant intracellular chlamydial inclusion morphology completely abrogated the τ₂-NAD(P)H increase inside the chlamydial inclusion. τ₂-NAD(P)H also decreased inside chlamydial inclusions when the cells were treated with IFNγ reflecting the reduced metabolism of persistent chlamydiae. Furthermore, a significant increase in τ₂-NAD(P)H and a decrease in the relative amount of free NAD(P)H inside the host cell nucleus indicated cellular starvation during intracellular chlamydial infection. Using FLIM analysis by two-photon microscopy we could visualize for the first time metabolic pathogen-host interactions during intracellular Chlamydia trachomatis infections with high spatial and temporal resolution in living cells. Our findings suggest that intracellular chlamydial metabolism is directly linked to cellular NAD(P)H signaling pathways that are involved in host cell survival and longevity.http://europepmc.org/articles/PMC3136453?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Márta Szaszák Philipp Steven Kensuke Shima Regina Orzekowsky-Schröder Gereon Hüttmann Inke R König Werner Solbach Jan Rupp |
spellingShingle |
Márta Szaszák Philipp Steven Kensuke Shima Regina Orzekowsky-Schröder Gereon Hüttmann Inke R König Werner Solbach Jan Rupp Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell. PLoS Pathogens |
author_facet |
Márta Szaszák Philipp Steven Kensuke Shima Regina Orzekowsky-Schröder Gereon Hüttmann Inke R König Werner Solbach Jan Rupp |
author_sort |
Márta Szaszák |
title |
Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell. |
title_short |
Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell. |
title_full |
Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell. |
title_fullStr |
Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell. |
title_full_unstemmed |
Fluorescence lifetime imaging unravels C. trachomatis metabolism and its crosstalk with the host cell. |
title_sort |
fluorescence lifetime imaging unravels c. trachomatis metabolism and its crosstalk with the host cell. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS Pathogens |
issn |
1553-7366 1553-7374 |
publishDate |
2011-07-01 |
description |
Chlamydia trachomatis is an obligate intracellular bacterium that alternates between two metabolically different developmental forms. We performed fluorescence lifetime imaging (FLIM) of the metabolic coenzymes, reduced nicotinamide adenine dinucleotides [NAD(P)H], by two-photon microscopy for separate analysis of host and pathogen metabolism during intracellular chlamydial infections. NAD(P)H autofluorescence was detected inside the chlamydial inclusion and showed enhanced signal intensity on the inclusion membrane as demonstrated by the co-localization with the 14-3-3β host cell protein. An increase of the fluorescence lifetime of protein-bound NAD(P)H [τ₂-NAD(P)H] inside the chlamydial inclusion strongly correlated with enhanced metabolic activity of chlamydial reticulate bodies during the mid-phase of infection. Inhibition of host cell metabolism that resulted in aberrant intracellular chlamydial inclusion morphology completely abrogated the τ₂-NAD(P)H increase inside the chlamydial inclusion. τ₂-NAD(P)H also decreased inside chlamydial inclusions when the cells were treated with IFNγ reflecting the reduced metabolism of persistent chlamydiae. Furthermore, a significant increase in τ₂-NAD(P)H and a decrease in the relative amount of free NAD(P)H inside the host cell nucleus indicated cellular starvation during intracellular chlamydial infection. Using FLIM analysis by two-photon microscopy we could visualize for the first time metabolic pathogen-host interactions during intracellular Chlamydia trachomatis infections with high spatial and temporal resolution in living cells. Our findings suggest that intracellular chlamydial metabolism is directly linked to cellular NAD(P)H signaling pathways that are involved in host cell survival and longevity. |
url |
http://europepmc.org/articles/PMC3136453?pdf=render |
work_keys_str_mv |
AT martaszaszak fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT philippsteven fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT kensukeshima fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT reginaorzekowskyschroder fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT gereonhuttmann fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT inkerkonig fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT wernersolbach fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell AT janrupp fluorescencelifetimeimagingunravelsctrachomatismetabolismanditscrosstalkwiththehostcell |
_version_ |
1725160938925981696 |