PMID- 27091298
OWN - NLM
STAT- MEDLINE
DCOM- 20170925
LR  - 20181113
IS  - 1872-8057 (Electronic)
IS  - 0303-7207 (Print)
IS  - 0303-7207 (Linking)
VI  - 429
DP  - 2016 Jul 5
TI  - Analysis of specific RNA in cultured cells through quantitative integration of 
      q-PCR and N-SIM single cell FISH images: Application to hormonal stimulation of 
      StAR transcription.
PG  - 93-105
LID - S0303-7207(16)30097-1 [pii]
LID - 10.1016/j.mce.2016.04.001 [doi]
AB  - The steroidogenic acute regulatory protein (StAR) has been proposed to serve as 
      the switch that can turn on/off steroidogenesis. We investigated the events that 
      facilitate dynamic StAR transcription in response to cAMP stimulation in MA-10 
      Leydig cells, focusing on splicing anomalies at StAR gene loci. We used 3' 
      reverse primers in a single reaction to respectively quantify StAR primary 
      (p-RNA), spliced (sp-RNA/mRNA), and extended 3' untranslated region (UTR) 
      transcripts, which were quantitatively imaged by high-resolution fluorescence in 
      situ hybridization (FISH). This approach delivers spatio-temporal resolution of 
      initiation and splicing at single StAR loci, and transfers individual mRNA 
      molecules to cytoplasmic sites. Gene expression was biphasic, initially showing 
      slow splicing, transitioning to concerted splicing. The alternative 3.5-kb mRNAs 
      were distinguished through the use of extended 3'UTR probes, which exhibited 
      distinctive mitochondrial distribution. Combining quantitative PCR and FISH 
      enables imaging of localization of RNA expression and analysis of RNA processing 
      rates.
CI  - Copyright (c) 2016 Elsevier Ireland Ltd. All rights reserved.
FAU - Lee, Jinwoo
AU  - Lee J
AD  - Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI 
      53706, United States; Endocrinology and Reproductive Physiology Program, 
      University of Wisconsin, Madison, WI 53706, United States. Electronic address: 
      jwlee7@wisc.edu.
FAU - Foong, Yee Hoon
AU  - Foong YH
AD  - Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI 
      53706, United States.
FAU - Musaitif, Ibrahim
AU  - Musaitif I
AD  - Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI 
      53706, United States.
FAU - Tong, Tiegang
AU  - Tong T
AD  - Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI 
      53706, United States.
FAU - Jefcoate, Colin
AU  - Jefcoate C
AD  - Department of Cell and Regenerative Biology, University of Wisconsin, Madison, WI 
      53706, United States; Endocrinology and Reproductive Physiology Program, 
      University of Wisconsin, Madison, WI 53706, United States; Molecular and 
      Environmental Toxicology Center, University of Wisconsin, Madison, WI 53706, 
      United States.
LA  - eng
GR  - R01 DK074819/DK/NIDDK NIH HHS/United States
GR  - R01 DK090249/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20160414
PL  - Ireland
TA  - Mol Cell Endocrinol
JT  - Molecular and cellular endocrinology
JID - 7500844
RN  - 0 (3' Untranslated Regions)
RN  - 0 (Phosphoproteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (steroidogenic acute regulatory protein)
RN  - 23583-48-4 (8-Bromo Cyclic Adenosine Monophosphate)
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - 3' Untranslated Regions/genetics
MH  - 8-Bromo Cyclic Adenosine Monophosphate/pharmacology
MH  - Animals
MH  - Base Pairing/genetics
MH  - Cells, Cultured
MH  - *Imaging, Three-Dimensional
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice
MH  - Microscopy/*methods
MH  - Mitochondria/drug effects/metabolism
MH  - Phosphoproteins/*genetics
MH  - Polyadenylation/drug effects/genetics
MH  - RNA/*metabolism
MH  - RNA Splicing/drug effects/genetics
MH  - RNA, Messenger/genetics/metabolism
MH  - Real-Time Polymerase Chain Reaction/*methods
MH  - Single-Cell Analysis/*methods
MH  - Time Factors
MH  - *Transcription, Genetic/drug effects
PMC - PMC6181224
MID - NIHMS987885
OTO - NOTNLM
OT  - Fluorescence in situ hybridization
OT  - PCR
OT  - Splicing
OT  - StAR
OT  - Transcription
EDAT- 2016/04/20 06:00
MHDA- 2017/09/26 06:00
PMCR- 2018/10/11
CRDT- 2016/04/20 06:00
PHST- 2015/12/31 00:00 [received]
PHST- 2016/04/02 00:00 [revised]
PHST- 2016/04/02 00:00 [accepted]
PHST- 2016/04/20 06:00 [entrez]
PHST- 2016/04/20 06:00 [pubmed]
PHST- 2017/09/26 06:00 [medline]
PHST- 2018/10/11 00:00 [pmc-release]
AID - S0303-7207(16)30097-1 [pii]
AID - 10.1016/j.mce.2016.04.001 [doi]
PST - ppublish
SO  - Mol Cell Endocrinol. 2016 Jul 5;429:93-105. doi: 10.1016/j.mce.2016.04.001. Epub 
      2016 Apr 14.