PMID- 10777692
OWN - NLM
STAT- MEDLINE
DCOM- 20000612
LR  - 20131121
IS  - 0006-291X (Print)
IS  - 0006-291X (Linking)
VI  - 271
IP  - 1
DP  - 2000 Apr 29
TI  - In situ RT-PCR detection of CYP1A mRNA in pharyngeal epithelium and chondroid 
      cells from chemically untreated fish: involvement in vertebrate craniofacial 
      skeletal development?
PG  - 130-7
AB  - Knowledge about the expression sites of cytochrome P450 1A (CYP1A) mRNA is 
      crucial for a better understanding of the physiological function of CYP1A. We 
      investigated the cellular localization of CYP1A mRNA in chemically untreated fish 
      by use of an in situ reverse transcription-polymerase chain reaction (IS RT-PCR) 
      technique. The fathead minnow (Pimephales promelas) was formalin-fixed, and 
      paraffin-embedded. Sections (5 microm) were treated with trypsinogen. Following 
      reverse transcription of CYP1A mRNA, the cDNA was amplified in situ by PCR with 
      specific primers. Detection of the amplicons was accomplished by a second PCR 
      performed with digoxigenin-labeled dUTP. CYP1A mRNA expression was detected in 
      cytoplasm of chondroid cells surrounding hyaline cartilage in gill arches. This 
      result was consistent with that of immunohistochemical analysis with a 
      CYP1A1-specific monoclonal antibody. CYP1A mRNA also was found in stratified 
      squamous epithelium of the pharynx and gill arches, but no staining was detected 
      in those cells by immunohistochemical analysis. The results indicate that IS 
      RT-PCR is an effective/sensitive technique for localizing low level CYP1A 
      expression. In addition, the sites where we identified expression of CYP1A are 
      targets of retinoic acid, sonic hedgehog and Hox genes, suggesting that 
      functional CYP1A in vertebrates could participate in craniofacial skeletal 
      development through involvement in the retinoic acid signaling cascade.
CI  - Copyright 2000 Academic Press.
FAU - Iwata, H
AU  - Iwata H
AD  - Biology Department, Woods Hole Oceanographic Institution, Woods Hole, 
      Massachusetts 02543, USA.
FAU - Stegeman, J J
AU  - Stegeman JJ
LA  - eng
GR  - P42ES07381/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (RNA, Messenger)
RN  - 9035-51-2 (Cytochrome P-450 Enzyme System)
RN  - NQ1SX9LNAU (Digoxigenin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Chondrocytes/*metabolism
MH  - Cloning, Molecular
MH  - Cytochrome P-450 Enzyme System/*genetics/physiology
MH  - Digoxigenin/metabolism
MH  - Epithelium/metabolism
MH  - Female
MH  - Fishes/embryology/*genetics
MH  - Immunohistochemistry
MH  - Liver/metabolism
MH  - Male
MH  - Molecular Sequence Data
MH  - Pharynx/*metabolism
MH  - RNA, Messenger/genetics
MH  - *Reverse Transcriptase Polymerase Chain Reaction
EDAT- 2000/04/25 09:00
MHDA- 2000/06/17 09:00
CRDT- 2000/04/25 09:00
PHST- 2000/04/25 09:00 [pubmed]
PHST- 2000/06/17 09:00 [medline]
PHST- 2000/04/25 09:00 [entrez]
AID - S0006-291X(00)92543-9 [pii]
AID - 10.1006/bbrc.2000.2543 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 2000 Apr 29;271(1):130-7. doi: 
      10.1006/bbrc.2000.2543.