Gestational profile of Na+/H+ exchanger and Cl-/HCO3- anion exchanger mRNA expression in placenta using real-time QPCR.
The onset of maternal blood flow (10-12 weeks gestation) results in increased oxygenation of the placenta. We investigated whether the expressions of Na+/H+ exchanger (NHE) and Cl-/HCO3- anion exchanger (AE), thought to have an important role in maintaining intracellular pH of the syncytiotrophoblast and fetal pH homeostasis, are altered at the same time as this increase in blood flow. Real-time quantitative PCR was used to examine steady state levels of NHE (NHE1, 2, 3) and AE (AE1, 2) mRNA expression in early (6-9 weeks) and late (10-13 weeks) first trimester and full-term (38-40 weeks) placentas. beta-Actin, IF2B and GAPDH mRNA was also measured. None of the genes showed a significant difference in expression between the early and late first trimester groups. However, NHE2 (p < 0.001) and GAPDH (p < 0.05) mRNA expression significantly increased 18- and 3.7-fold between early first trimester and term. In conclusion, this study provides additional evidence that GAPDH is an unsuitable housekeeping gene for normalization of transcript levels in placenta. The expression of NHE and AE in the villous placenta is not altered concomitant with the onset of maternal blood flow. However, NHE2 transcripts appear to be gestationally regulated, which may contribute to changes in NHE activity.
Lacey
H A
HA
Academic Unit of Child Health, University of Manchester, St. Mary's Hospital, Hathersage Road, Manchester M13 0JH, UK. mdsgsh12@man.ac.uk
Nolan
T
T
Greenwood
S L
SL
Glazier
J D
JD
Sibley
C P
CP
D000328:Adult; D027963:Chloride-Bicarbonate Antiporters / Q000235:genetics / Q000378:metabolism*; D002824:Chorionic Villi / Q000378:metabolism*; D005260:Female; D015870:Gene Expression; D020869:Gene Expression Profiling; D005865:Gestational Age; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics / Q000378:metabolism*
D027963:Chloride-Bicarbonate Antiporters; D012333:RNA, Messenger; C081517:SLC9A2 protein, human; D017923:Sodium-Hydrogen Exchangers; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating)