PMID- 32944909
OWN - NLM
STAT- MEDLINE
DCOM- 20210330
LR  - 20210330
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2214
DP  - 2021
TI  - Investigating the Inner Cell Mass of the Mouse Blastocyst by Combined 
      Immunofluorescence Staining and RNA Fluorescence In Situ Hybridization.
PG  - 157-173
LID - 10.1007/978-1-0716-0958-3_11 [doi]
AB  - Immunofluorescence and RNA fluorescence in situ hybridization (FISH) methods 
      enable the detection of, respectively, proteins and RNA molecules in single 
      cells. Adapted to preimplantation mouse embryos, these techniques allow the 
      investigation of transcriptional dynamics in the first embryonic and 
      extraembryonic lineages and can circumvent the limited amount of starting 
      material. This can as well be coupled to examination of chromatin modification, 
      i.e., histone marks, by immunofluorescence. Here is outlined an 
      immunofluorescence protocol combined to nascent RNA-FISH after immunosurgery of 
      the mouse inner cell mass of the blastocyst to study early changes in 
      transcription and/or histone marks of both primitive endoderm and epiblast cells. 
      The method describes the different steps from coverslips and FISH probe 
      preparation to inner cell mass isolation and immunofluorescence followed by 
      RNA-FISH. Furthermore, this is applicable to earlier developmental stages and 
      other mammalian species provided little technical adjustments.
FAU - Borensztein, Maud
AU  - Borensztein M
AD  - Institut Curie, CNRS UMR3215/ INSERM U934, Paris Sciences & Lettres Research 
      University (PSL), Paris, France. maud.borensztein@curie.fr.
AD  - Institut de Genetique Moleculaire de Montpellier, Univ Montpellier, CNRS, 
      Montpellier, France. maud.borensztein@curie.fr.
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 63231-63-0 (RNA)
SB  - IM
MH  - Animals
MH  - Blastocyst/*cytology/metabolism
MH  - Endoderm/cytology/metabolism
MH  - Fluorescent Antibody Technique/*methods
MH  - Germ Layers/cytology/metabolism
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice/*embryology/genetics
MH  - RNA/*analysis/genetics
MH  - Transcription, Genetic
OTO - NOTNLM
OT  - IF /RNA-FISH
OT  - Immunosurgery
OT  - Inner cell mass
OT  - Nascent transcripts
OT  - Preimplantation embryo
EDAT- 2020/09/19 06:00
MHDA- 2021/03/31 06:00
CRDT- 2020/09/18 05:53
PHST- 2020/09/18 05:53 [entrez]
PHST- 2020/09/19 06:00 [pubmed]
PHST- 2021/03/31 06:00 [medline]
AID - 10.1007/978-1-0716-0958-3_11 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2021;2214:157-173. doi: 10.1007/978-1-0716-0958-3_11.