PMID- 26184691
OWN - NLM
STAT- MEDLINE
DCOM- 20160706
LR  - 20181202
IS  - 1549-4918 (Electronic)
IS  - 1066-5099 (Linking)
VI  - 33
IP  - 10
DP  - 2015 Oct
TI  - Single Cell Analysis Reveals Concomitant Transcription of Pluripotent and Lineage 
      Markers During the Early Steps of Differentiation of Embryonic Stem Cells.
PG  - 2949-60
LID - 10.1002/stem.2108 [doi]
AB  - The differentiation of embryonic stem cells is associated with extensive changes 
      in gene expression. It is not yet clear whether these changes are the result of 
      binary switch-like mechanisms or that of continuous and progressive variation. 
      Here, I have used immunostaining and single molecule RNA fluorescence in situ 
      hybridization (FISH) to assess changes in the expression of the well-known 
      pluripotency-associated gene Pou5f1 (also known as Oct4) and early 
      differentiation markers Sox1 and T-brachyury in single cells during the early 
      steps of differentiation of mouse embryonic stem cells. I found extensive overlap 
      between the expression of Pou5f1/Sox1 or Pou5f1/T-brachyury shortly after the 
      initiation of differentiation towards either the neuronal or the mesendodermal 
      lineage, but no evidence of correlation between their respective expression 
      levels. Quantitative analysis of transcriptional output at the sites of nascent 
      transcription revealed that Pou5f1 and Sox1 were transcribed in pulses and that 
      embryonic stem cell differentiation was accompanied by changes in pulsing 
      frequencies. The progressive induction of Sox1 was further associated with an 
      increase in the average size of individual transcriptional bursts. Surprisingly, 
      single cells that actively and simultaneously transcribe both the pluripotency- 
      and the lineage-associated genes could easily be found in the differentiating 
      population. The results presented here show for the first time that lineage 
      priming can occur in cells that are actively transcribing a pluripotent marker. 
      Furthermore, they suggest that this process is associated with changes in 
      transcriptional dynamics.
CI  - (c) 2015 AlphaMed Press.
FAU - Lanctot, Christian
AU  - Lanctot C
AD  - Institute of Cellular Biology and Pathology, First Faculty of Medicine, Charles 
      University in Prague, Prague, Czech Republic.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Stem Cells
JT  - Stem cells (Dayton, Ohio)
JID - 9304532
RN  - 0 (Fetal Proteins)
RN  - 0 (Octamer Transcription Factor-3)
RN  - 0 (Pou5f1 protein, mouse)
RN  - 0 (SOXB1 Transcription Factors)
RN  - 0 (Sox1 protein, mouse)
RN  - 0 (T-Box Domain Proteins)
RN  - EQ43SC3GDB (Brachyury protein)
SB  - IM
MH  - Animals
MH  - Cell Differentiation/*genetics
MH  - Cell Lineage/genetics
MH  - Embryonic Stem Cells/cytology/*metabolism
MH  - Fetal Proteins/biosynthesis/genetics
MH  - Gene Expression Regulation, Developmental
MH  - In Situ Hybridization, Fluorescence
MH  - Mice
MH  - Octamer Transcription Factor-3/biosynthesis/genetics
MH  - Pluripotent Stem Cells/cytology/*metabolism
MH  - SOXB1 Transcription Factors/biosynthesis/genetics
MH  - *Single-Cell Analysis
MH  - T-Box Domain Proteins/biosynthesis/genetics
OTO - NOTNLM
OT  - Early differentiation
OT  - Embryonic stem cells
OT  - Lineage priming
OT  - Single molecule RNA fluorescence in situ hybridization
OT  - Transcriptional dynamics
EDAT- 2015/07/18 06:00
MHDA- 2016/07/07 06:00
CRDT- 2015/07/18 06:00
PHST- 2014/12/09 00:00 [received]
PHST- 2015/05/04 00:00 [revised]
PHST- 2015/06/10 00:00 [accepted]
PHST- 2015/07/18 06:00 [entrez]
PHST- 2015/07/18 06:00 [pubmed]
PHST- 2016/07/07 06:00 [medline]
AID - 10.1002/stem.2108 [doi]
PST - ppublish
SO  - Stem Cells. 2015 Oct;33(10):2949-60. doi: 10.1002/stem.2108.