Oxygen-dependent modulation of erythropoietin mRNA levels in isolated rat kidneys studied by RNase protection.
Using oligonucleotide primers complementary to conserved regions in the mouse erythropoietin (Epo) gene, a portion of the rat Epo gene was amplified by the polymerase chain reaction to produce a probe suitable for assay of rat Epo mRNA by RNAse protection. The assay, which has sufficient sensitivity to measure to Epo mRNA in unstimulated rat kidneys, was used to demonstrate high amplitude in vitro modulation of Epo mRNA levels in response to changes in perfusate flow rate and oxygen tension in isolated kidneys, thus providing clear evidence that all the necessary events linking changes in oxygen delivery to the modulation of Epo mRNA levels can occur intrarenally.
Ratcliffe
P J
PJ
Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
Jones
R W
RW
Phillips
R E
RE
Nicholls
L G
LG
Bell
J I
JI
D000818:Animals; D004921:Erythropoietin / Q000235:genetics*; D005786:Gene Expression Regulation / Q000187:drug effects*; D066298:In Vitro Techniques; D007668:Kidney / Q000378:metabolism*; D008297:Male; D015345:Oligonucleotide Probes; D010100:Oxygen / Q000494:pharmacology*; D010313:Partial Pressure; D010477:Perfusion; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012260:Ribonucleases; D014158:Transcription, Genetic
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004921:Erythropoietin; D012260:Ribonucleases; D010100:Oxygen