PMID- 23749471
OWN - NLM
STAT- MEDLINE
DCOM- 20140317
LR  - 20130821
IS  - 1097-0177 (Electronic)
IS  - 1058-8388 (Linking)
VI  - 242
IP  - 9
DP  - 2013 Sep
TI  - mRNA fluorescence in situ hybridization to determine overlapping gene expression 
      in whole-mount mouse embryos.
PG  - 1094-100
LID - 10.1002/dvdy.23993 [doi]
AB  - BACKGROUND: Whole-mount in situ hybridization (ISH) is a prevalent tool to 
      examine the spatial distribution of gene transcripts in intact embryos. 
      Chromogenic-based methods of signal development are commonly used in mouse 
      embryos because of their high sensitivity. Fluorescence techniques, however, 
      offer several advantages over chromogenic methods including the ability to 
      visualize multiple signals in a specimen at once. RESULTS: We describe a 
      procedure for fluorescence in situ hybridization (FISH) for whole mouse embryos 
      up to embryonic day 13.5. We show that this approach successfully produces a 
      bright expression signal for several genes, validating the procedure in multiple 
      tissues. Further, we show that double FISH can be used to visualize the 
      expression of two genes in a single embryo by determining that Hoxd13 and Shh are 
      co-expressed in both the limb bud and the hindgut. Finally, we demonstrate that 
      FISH can be paired with confocal microscopy to take optical sections of interior 
      regions of the embryo. CONCLUSIONS: FISH is a valid alternative to 
      chromogenic-based ISH for visualizing gene expression in whole mouse embryos. 
      This work provides a framework to add additional fluorescence signals in the 
      mouse such as visualizing both mRNA and protein by pairing the procedure with 
      immunofluorescence.
CI  - Copyright (c) 2013 Wiley Periodicals, Inc.
FAU - Neufeld, Stanley J
AU  - Neufeld SJ
AD  - Department of Biological Sciences, University of Calgary, Calgary, Canada.
FAU - Zhou, Xiaolan
AU  - Zhou X
FAU - Vize, Peter D
AU  - Vize PD
FAU - Cobb, John
AU  - Cobb J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130627
PL  - United States
TA  - Dev Dyn
JT  - Developmental dynamics : an official publication of the American Association of 
      Anatomists
JID - 9201927
RN  - 0 (Hedgehog Proteins)
RN  - 0 (Homeodomain Proteins)
RN  - 0 (Hoxd13 protein, mouse)
RN  - 0 (RNA, Messenger)
RN  - 0 (Shh protein, mouse)
RN  - 0 (Transcription Factors)
SB  - IM
MH  - Animals
MH  - Embryo, Mammalian/cytology/*metabolism
MH  - Gene Expression Regulation, Developmental/*physiology
MH  - Hedgehog Proteins/biosynthesis
MH  - Hindlimb/cytology/embryology
MH  - Homeodomain Proteins/biosynthesis
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Intestines/cytology/embryology
MH  - Mice
MH  - RNA, Messenger/*biosynthesis
MH  - Transcription Factors/biosynthesis
OTO - NOTNLM
OT  - Hoxd13
OT  - Shh
OT  - co-expression
OT  - fluorescence in situ hybridization
OT  - limb development
OT  - mouse
OT  - whole-mount
EDAT- 2013/06/12 06:00
MHDA- 2014/03/19 06:00
CRDT- 2013/06/11 06:00
PHST- 2013/01/24 00:00 [received]
PHST- 2013/05/24 00:00 [revised]
PHST- 2013/05/25 00:00 [accepted]
PHST- 2013/06/11 06:00 [entrez]
PHST- 2013/06/12 06:00 [pubmed]
PHST- 2014/03/19 06:00 [medline]
AID - 10.1002/dvdy.23993 [doi]
PST - ppublish
SO  - Dev Dyn. 2013 Sep;242(9):1094-100. doi: 10.1002/dvdy.23993. Epub 2013 Jun 27.