Abstract
The kidney is a main site of erythropoietin production in the body. We developed a new method for the detection of Epo protein by deglycosylation-coupled Western blotting. Detection of deglycosylated Epo enables the examination of small changes in Epo production. Using this method, we investigated the effects of angiotensin II (ATII) on Epo production in the kidney. ATII stimulated the plasma Epo concentration; Epo, HIF2α, and PHD2 mRNA expression in nephron segments in the renal cortex and outer medulla; and Epo protein expression in the renal cortex. In situ hybridization and immunohistochemistry revealed that ATII stimulates Epo mRNA and protein expression not only in proximal tubules but also in collecting ducts, especially in intercalated cells. These data support the regulation of Epo production in the kidney by the renin–angiotensin–aldosterone system (RAS).
Highlights
Erythropoietin (Epo) stimulates erythrocyte production in the bone marrow [1]
Severe hypoxia and severe anemia cause more than a 500-fold increase in the serum Epo concentration [5]
We developed a new Western blotting method and found that deglycosylated Epo is an accurate marker of Epo expression [5,9]
Summary
Hypoxia and anemia stimulate Epo production by the kidney [1,2,3,4]. Severe hypoxia and severe anemia cause more than a 500-fold increase in the serum Epo concentration [5]. Such conditions are rare and most people do not experience them during their lives. The serum Epo concentration in normal people is low, and Epo protein expression in control kidneys has been too low to detect [5,6,7,8]. The detection of small changes in Epo production has become possible
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