Quantitation of the mRNA levels of Epo and EpoR in various tissues in the ovine fetus.
A partial cDNA of the sheep erythropoietin receptor (EpoR) was obtained and used in real-time PCR to quantitate mRNA levels in placenta, liver and kidney throughout development (term=150 days). This was compared with Epo mRNA levels in the same tissues. Both Epo and EpoR mRNA were present in the placenta throughout gestation at low levels from 66 days onwards and these did not vary throughout gestation. Compared with the expression levels in the placenta, the levels of EpoR gene expression in the liver at 66, 99 and 140 days were, median (range)-288 (120-343), 278 (63-541) and 7 (3-15), respectively, reflecting the disappearance of erythropoiesis after 130 days. Low levels of EpoR gene expression were seen in the kidney at 3 (2-5), 5 (2-7), and 7 (2-10) times that in the placenta at 66, 99, and 140 days, respectively. By hybridization histochemistry the EpoR mRNA was located in the proximal tubular cells of the mesonephros and metanephros at 42 days. Epo mRNA levels in the kidney were 215 (116-867), 528 (113-765) and 46 (15-204) times those in the placenta at 69, 99, and 140 days, respectively. In the liver at the same ages the concentrations of mRNA were lower than in the kidney, the liver/placenta ratios being 50 (11-90), 17 (3-39), 9 (5-14). At 130 days Epo/EpoR levels in the hippocampus were 6+/-3 and 8+/-3 times that in the term placenta, respectively. These studies demonstrate that the ovine placenta expresses the Epo gene from at least 66 days of gestation. However, gene expression levels are very low compared with those in the liver and kidney, and even the hippocampus.
David
R Bruce
RB
Norwegian School of Veterinary Science, Oslo, Norway
Lim
Gaik Bee
GB
Moritz
Karen M
KM
Koukoulas
Irene
I
Wintour
E Marelyn
EM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004921:Erythropoietin / Q000235:genetics* / Q000378:metabolism; D005260:Female; D005333:Fetus / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects*; D006624:Hippocampus / Q000378:metabolism; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000196:embryology / Q000378:metabolism; D008099:Liver / Q000196:embryology / Q000378:metabolism; D008969:Molecular Sequence Data; D009929:Organ Size; D010920:Placenta / Q000196:embryology / Q000378:metabolism; D011247:Pregnancy; D012333:RNA, Messenger / Q000378:metabolism*; D017467:Receptors, Erythropoietin / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012756:Sheep / Q000196:embryology*
D012333:RNA, Messenger; D017467:Receptors, Erythropoietin; D004921:Erythropoietin