10.5061/DRYAD.S04BB
Featherstone, Karen
University of Manchester
Hey, Kirsty
University of Warwick
Momiji, Hiroshi
University of Warwick
McNamara, Anne V.
University of Manchester
Patist, Amanda L.
University of Manchester
Woodburn, Joanna
University of Manchester
Spiller, David G.
University of Manchester
Christian, Helen C.
University of Oxford
McNeilly, Alan S.
University of Edinburgh
Mullins, John J.
University of Edinburgh
Finkenstadt, Barbel F.
University of Warwick
Rand, David A.
University of Warwick
White, Michael R. H.
University of Manchester
Davis, Julian R. E.
University of Manchester
Data from: Spatially coordinated dynamic gene transcription in living
pituitary tissue
Dryad
dataset
2016
live-cell microscopy
Transcription Dynamics
Spatial Organisation
stochastic modelling
2016-02-11T15:58:40Z
2016-02-11T15:58:40Z
en
https://doi.org/10.7554/eLife.08494
11717459 bytes
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CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
Transcription at individual genes in single cells is often pulsatile and
stochastic. A key question emerges regarding how this behaviour
contributes to tissue phenotype, but it has been a challenge to
quantitatively analyse this in living cells over time, as opposed to
studying snap-shots of gene expression state. We have used imaging of
reporter gene expression to track transcription in living pituitary
tissue. We integrated live-cell imaging data with statistical modelling
for quantitative real-time estimation of the timing of switching between
transcriptional states across a whole tissue. Multiple levels of
transcription rate were identified, indicating that gene expression is not
a simple binary ‘on-off’ process. Immature tissue displayed shorter
durations of high-expressing states than the adult. In adult pituitary
tissue, direct cell contacts involving gap junctions allowed local spatial
coordination of prolactin gene expression. Our findings identify how
heterogeneous transcriptional dynamics of single cells may contribute to
overall tissue behaviour.
Adult Pituitary Data1GFP fluorescence measured from single cells in adult
pituitary tissue and subsequent modelling of transcription activity using
a stochastic switch model.Adult Pituitary Data2GFP fluorescence measured
from single cells in adult pituitary tissue and subsequent modelling of
transcription activity using a stochastic switch model.Adult Pituitary
Data3GFP fluorescence measured from single cells in adult pituitary tissue
and subsequent modelling of transcription activity using a stochastic
switch model.E18_5 Pituitary Data1GFP fluorescence measured from single
cells in embryonic day18.5 pituitary tissue and subsequent modelling of
transcription activity using a stochastic switch model.E18_5 Pituitary
Data2GFP fluorescence measured from single cells in embryonic day18.5
pituitary tissue and subsequent modelling of transcription activity using
a stochastic switch model.P1_5 Pituitary Data1GFP fluorescence measured
from single cells in post-natal day1.5 pituitary tissue and subsequent
modelling of transcription activity using a stochastic switch model.P1_5
Pituitary Data2GFP fluorescence measured from single cells in post-natal
day1.5 pituitary tissue and subsequent modelling of transcription activity
using a stochastic switch model.Adult Pituitary Trypsin Untreated Data1GFP
fluorescence measured from single cells in adult pituitary tissue. Control
sample to compare to Adult Pituitary Trypsin Treated Data1.Adult Pituitary
Trypsin Treated Data1GFP fluorescence measured from single cells in adult
pituitary tissue treated with trypsin for 2hrs prior to confocal imaging.
Compare to Adult Pituitary Trypsin Untreated Data1.Adult Pituitary Trypsin
Untreated Data2GFP fluorescence measured from single cells in adult
pituitary tissue. Control sample to compare to Adult Pituitary Trypsin
Treated Data2.Adult Pituitary Trypsin Treated Data2GFP fluorescence
measured from single cells in adult pituitary tissue treated with trypsin
for 2hrs prior to confocal imaging. Compare to Adult Pituitary Trypsin
Untreated Data2.Adult Pituitary_Cx_AGA_Data1GFP fluorescence measured from
single cells in adult pituitary tissue treated with 18alpha-glycyrrhetinic
acid (20uM) for 2hrs prior to confocal imaging. Compare to Adult Pituitary
Data2.Adult Pituitary_Cx_AGA_Data2GFP fluorescence measured from single
cells in adult pituitary tissue treated with 18alpha-glycyrrhetinic acid
(20uM) for 2hrs prior to confocal imaging. Compare to Adult Pituitary
Data3.