PMID- 30414154
OWN - NLM
STAT- MEDLINE
DCOM- 20190605
LR  - 20231110
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Print)
IS  - 1064-3745 (Linking)
VI  - 1890
DP  - 2019
TI  - Quantifying Tissue-Specific Overexpression of FOXO in Drosophila via mRNA 
      Fluorescence In Situ Hybridization Using Branched DNA Probe Technology.
PG  - 171-190
LID - 10.1007/978-1-4939-8900-3_15 [doi]
AB  - While the highly conserved FOXO transcription factors have been studied in 
      Drosophila melanogaster for decades, the ability to accurately control and 
      measure their tissue-specific expression is often cumbersome due to a lack of 
      reagents and to limited, nonhomogeneous samples. The need for quantitation within 
      a distinct cell type is particularly important because transcription factors must 
      be expressed in specific amounts to perform their functions properly. However, 
      the inherent heterogeneity of many samples can make evaluating cell-specific FOXO 
      and/or FOXO load difficult. Here, we describe an extremely sensitive fluorescence 
      in situ hybridization (FISH) approach for visualizing and quantifying multiple 
      mRNAs with single-cell resolution in adult Drosophila cardiomyocytes. The 
      procedure relies upon branched DNA technology, which allows several fluorescent 
      molecules to label an individual transcript, drastically increasing the 
      signal-to-noise ratio compared to other FISH assays. This protocol can be 
      modified for use in various small animal models, tissue types, and for assorted 
      nucleic acids.
FAU - Blice-Baum, Anna C
AU  - Blice-Baum AC
AD  - Division of Cardiology, Department of Medicine, Johns Hopkins University School 
      of Medicine, Baltimore, MD, USA. anna.c.blice-baum@cabrini.edu.
AD  - Science Department, Iadarola Center for Science, Education and Technology, 
      Cabrini University, Radnor, PA, USA. anna.c.blice-baum@cabrini.edu.
FAU - Vogler, Georg
AU  - Vogler G
AD  - Development, Aging and Regeneration Program, Sanford Burnham Prebys Medical 
      Discovery Institute, La Jolla, CA, USA. gvogler@sbpdiscovery.org.
FAU - Viswanathan, Meera C
AU  - Viswanathan MC
AD  - Division of Cardiology, Department of Medicine, Johns Hopkins University School 
      of Medicine, Baltimore, MD, USA.
FAU - Trinh, Bosco
AU  - Trinh B
AD  - Development, Aging and Regeneration Program, Sanford Burnham Prebys Medical 
      Discovery Institute, La Jolla, CA, USA.
FAU - Limpitikul, Worawan B
AU  - Limpitikul WB
AD  - Department of Biomedical Engineering, Johns Hopkins University School of 
      Medicine, Baltimore, MD, USA.
FAU - Cammarato, Anthony
AU  - Cammarato A
AD  - Division of Cardiology, Department of Medicine, Johns Hopkins University School 
      of Medicine, Baltimore, MD, USA.
AD  - Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, 
      MD, USA.
LA  - eng
GR  - R01 HL124091/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (DNA Probes)
RN  - 0 (Forkhead Transcription Factors)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Animals
MH  - *DNA Probes
MH  - Drosophila melanogaster
MH  - Forkhead Transcription Factors/*genetics
MH  - *Gene Expression
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Microscopy, Fluorescence
MH  - Myocardium/metabolism
MH  - Organ Specificity/genetics
MH  - RNA, Messenger/*genetics
PMC - PMC7906431
MID - NIHMS1667824
OTO - NOTNLM
OT  - Branched DNA
OT  - Dorsal vessel
OT  - Drosophila melanogaster
OT  - FISH
OT  - Fluorescence in situ hybridization
OT  - Heart tube
OT  - RNAscope
OT  - ViewRNA
OT  - bDNA
EDAT- 2018/11/11 06:00
MHDA- 2019/06/06 06:00
PMCR- 2021/02/25
CRDT- 2018/11/11 06:00
PHST- 2018/11/11 06:00 [entrez]
PHST- 2018/11/11 06:00 [pubmed]
PHST- 2019/06/06 06:00 [medline]
PHST- 2021/02/25 00:00 [pmc-release]
AID - 10.1007/978-1-4939-8900-3_15 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2019;1890:171-190. doi: 10.1007/978-1-4939-8900-3_15.