A method of coculturing adult rat hepatic parenchymal cells (PC) and stromal cells in a three-dimensional framework of nylon filtration screens or biodegradable polymer meshes was developed in our laboratory. Rat liver stroma, which includes vascular and bile duct endothelial cells, fat-storing cells, fibroblasts, and Kupffer cells, were isolated by gradient centrifugation after in situ liver perfusion and expanded in monolayer culture prior to seeding onto nylon screens or bioresorbable polyglycolic acid (PGA) polymers oriented into a felt-like construct. A second inoculum of freshly isolated PC was applied after the stromal cells became established. Histological analyses revealed that PC proliferation occurred until all available space for expansion within the template was exhausted. These cells retained their rounded morphology, and after 4-5 wk 7-9 "layers" of PC filled the 140-microns deep template. Dioxin-inducible cytochrome P450 activity was detected for up to 58 d in culture, and albumin, fibrinogen, transferrin, and soluble fibronectin were detected in the medium by enzyme-linked immunosorbent assay (ELISA) for 48 d in vitro. Immunohistochemical analysis of sections through the cultures confirmed the presence of these proteins as well as cytokeratin at the cellular level; the extracellular matrix stained for both collagen type III and laminin. Long-term PC proliferation and function were enhanced by the presence of stromal cells as well as by a meshwork template whose geometry allows the interaction of PC with stroma and matrix on several different planes. To permit transplantation, cocultures of hepatic PC and stromal cells were established on PGA felt constructs instead of nylon screens. After approximately 24 d in vitro, these constructs were grafted into sites in the mesentery, omentum, and subcutaneous tissues of adult Long-Evans rats. The growth of hepatocytes after 30 d in situ was evident by histological analysis; grafts of cocultures regenerated a liver-like architecture consisting of sinusoids and putative biliary structures. In addition, PC at these extrahepatic graft sites were positive for albumin, transferrin, and fibrinogen synthesis by immunohistochemistry. Graft survival was enhanced when recipients were subjected to approximately 40% hepatectomy. Hepatic PC:stromal cell cocultures may prove useful in the restoration of liver function either by direct transplantation using PGA or similar templates, or as extracorporeal devices, using nylon screens.

Light microscopic, scanning electron microscopic, and transmission electron microscopic studies of the early developmental stages of chick embryonic bone marrow disclose characteristic associations of the first hematopoietic cells with stromal cells. The first hematopoietic cells, large basophilic cells that we have termed presumptive stem cells, segregate into erythropoietic and granulopoietic regions. Intravascular erythropoietic cells associate with sinusoidal endothelial cells, while granulopoietic cells associate with extravascular reticular cells. Extensive, intimate contacts between erythroid and endothelial cells are maintained, in part, by marginal arrays of microtubules, which promote a flattening of the adherent erythroid cell surface. In addition, cell surface components of opposing cells, visualized by ruthenium red staining, appear to merge and possibly to interact. Granulopoietic cells establish intimate but less extensive associations with reticular cells through cell-surface interactions. Stationary granuloid cells appear to be held in place by small, thin processes emanating from the sheet-like reticular cells. Granuloid cells are capable of moving within the extravascular region, using reticular cell surfaces as a substrate. Intimate associations also occur among granulopoietic cells, the significance of which is unclear. Thus, sinusoidal endothelial cells and reticular cells comprise the critical non-hematopoietic or stromal elements of avian bone marrow, where they have a putative role in segregating presumptive stem cells into erythrocyteic and granulocytic compartments. They serve as an architectual, and possibly regulatory, framework on which hematopoiesis occurs.

Rats were rendered anaemic by a single bleeding or by a single injection of phenylhydrazine. At various times after the onset of anaemia they were nephrectomized and challenged with a 6 h exposure to hypoxia. The erythropoietin titre observed at the end of this hypoxic period was corrected for renal erythropoietin induced by the anaemia alone, and the resulting extrarenal component was compared to total erythropoietin production of nephric rats in response to anaemia plus 6 h hypoxia. Extrarenal erythropoietin production was found to increase from 10.3% in normal rats to 12.5% in moderately anaemic rats to 15.1% in rats with severe bleeding anaemia. In phenylhydrazine-treated rats this extrarenal component was found to be 18.3% possibly due to stimulation of extrarenal erythropoietin by haemolysed red cells. Chronic phenylhydrazine administration resulted in splenomegaly and Kupffer cell hyperactivity but not in any further stimulation of extrarenal erythropoietin production.

The principal function of erythrocytes is the transport of oxygen. Erythropoiesis proceeds at a rate consistent with the demand for oxygen-carrying capacity, and the major regulator of erythrocyte production is erythropoietin. Erythropoietin is produced primarily by the kidney under control of a tissue oxygenation sensor. The recently developed erythropoietin radioimmunoassay should provide a clinically useful tool. Erythrocytosis is a pathologic state characterized by an elevated erythrocyte mass, which may result from increased proliferation of erythroid progenitors due to an intrinsic cellular defect or in response to extrinsic signals. Secondary erythrocytosis results from either physiologically appropriate compensation for inadequate tissue oxygenation or from inappropriate stimulation of erythropoiesis. Erythrocytosis increases oxygen-carrying capacity of the blood, but at high hematocrit levels increased blood viscosity may result in decreased tissue oxygen delivery. Polycythemia vera is a hematopoietic stem cell disease of clonal origin. Initial results from the Polycythemia Rubra Study Group suggest that therapy with chlorambucil is associated with an unacceptably high risk for development of acute leukemia, and 32P is preferred for situations in which phlebotomy alone is insufficient.

Double partial hepatectomy (hepx) evokes an elevation in serum erythropoietin (Ep) levels in anephric hypoxic animals when compared to non-hypoxic or sham hepx controls. But this Ep response is significantly lower than that found in singly hepx, anephric hypoxic rats. Double hepx also induces numerous cytological changes in the liver. Extravascular accumulation of fat, fibrous scarring, localized necroses, and multiple abscesses, as well as decreased vascularity, occur following the second hepx. A humoral factor was detected in the serum of these animals that is capable of inducing hepatic Ep production when injected into normal rats 18 hours before nephrectomy and hypoxia. This factor, termed hepatopoietin (Hp), was previously demonstrated in the venous serum of singly hepx rats. The serum from animals subjected to double partial hepx is not as potent in inducing Ep production as the serum from singly hepx animals. The discrepancies noted between the single and double hepx groups is attributed to the necrotic cytological changes described above.

The site of erythropoietin (Ep) production and/or storage in the rat liver was determined. A guinea pig anti-Ep was produced against purified rat Ep (64,096 +/- 4j064 IU/mg). This antibody was found to be highly specific using rocket immunoelectrophoresis, Ouchterlony gel diffusion methods, and immunoprecipitin reactions as well as Ep neutralization tests (capable of completely neutralizing up to 2,000 IU Ep/mg). This anti-Ep was labeled with either fluorescein for light microscopic study or ferritin for electron microscopy. Kupffer cells showed varying degrees of labeling after hepatectomy alone or hepatectomy combined with nephrectomy and/or hypoxia. Greatest labeling was seen in Kupffer cells of rats that were nephrectomized 48 hr posthepatectomy and kept at ambient pressure. No labeling of hepatocytes or vascular and bile duct endothelium was noted.

The development of a cell culture system that produces erythropoietin (Epo) in a regulated manner has been the focus of much effort. We have screened multiple renal and hepatic cell lines (including MDCK, LLC-PK1, BHK, WRL 68, CLCL, A704, CRFK, A498, ACHN, TCMK-1, LLC-MK2, CaKi-2, HepG2, and Hep3B) for either constitutive or regulated expression of Epo. Only the human hepatoma cell lines, Hep3B and HepG2, made significant amounts of Epo as measured both by radioimmunoassay and in vitro bioassay (as much as 330 milliunits per 10(6) cells in 24 hr). The constitutive production of Epo increased dramatically as a function of cell density in both cell lines. At cell densities less than 3.3 X 10(5) cells per cm2, there was little constitutive release of Epo in the medium (less than 30 milliunits per 10(6) cells in 24 hr). With Hep3B cells grown at low cell densities, a mean 18-fold increase in Epo expression was seen in response to hypoxia and a 6-fold increase was observed in response to incubation in medium containing 50 microM cobalt(II) chloride. At similar low cell densities, Epo production in HepG2 cells could be enhanced an average of about 3-fold by stimulation with either hypoxia or cobalt(II) chloride. Upon such stimulation, both cell lines demonstrated markedly elevated levels of Epo mRNA. Hence, both Hep3B and HepG2 cell lines provide an excellent in vitro system in which to study the physiological regulation of Epo expression.

Hepatic cells in rats were evaluated after subtotal hepatectomy using scintillation scanning with technetium sulfur colloid (TSC), autoradiography, and microstereology techniques. The ability of the liver to accumulate TSC increased during the course of the regeneration as did the labeling of Kupffer and parenchymal cells with tritiated thymidine (3H-tdR). Kupffer to parenchymal cell number ratios and Kupffer cell relative areas were also elevated, attaining peak values at 72 hours post-hepatectomy. This period corresponds to the time of peak erythropoietin (Ep) production in rats with regenerating livers after nephrectomy and exposure to hypoxia. These findings suggest that the Kupffer cell may function as a cellular site of Ep formation.

Erythropoietin (Ep) is a glycoprotein hormone that is responsible for mammalian red blood cell production. Adult rat liver regenerating 48-72 h after hepatectomy (hepx) produces elevated levels of Ep in response to hypoxia when compared to sham-operated, anephric hypoxic controls. A factor, termed hepatopoietin (Hp), found in the serum of hepx rats, is capable of stimulating hepatic Ep production when administered to normal rats 18 h prior to hypoxic exposure. Although the hepatic vein is the most potent source of this factor, Hp can also be demonstrated in the systemic arterial circulation. Bilateral nephrectomy (nephrx) of the donor hepx animal 24 h prior to bleeding abolishes this variation, and highest Ep levels are noted when serum from a hepx and nephrx rat is administered to animals immediatley after nephrx and 18 h before hypoxic exposure. Serum derived from hepx male rats displays a greater ability to evoke hepatic Ep production in normal recipients than serum from similarly treated female rats. Regardless of the sex of the hepx donor, Ep elaboration after hypoxia is highest in male recipients. The results indicate that there is a sexual variation in the production of Hp as well as Ep.

Prostaglandins A2, E1, E2, methylated E2s, and F2 alpha affected erythropoiesis and/or erythropoietin (Ep) production. This action is indicated in the exhypoxic, polycythemic mouse where radioiron incorporations into RBC increased after administration of these compounds. The kidney and liver have been indicated through previous studies, to actively participate in Ep production. The kidney and liver have been indicated through previous studies, to actively participate in Ep production. By the removal of one of these active sites in a murine system treated with prostaglandins it is shown that a response is reflected in Ep levels. Interference of the action of prostaglandins (PG) is altered by the removal of these target sites of Ep production. The erythropoietic responses elicited by PGA2, E1, and perhaps the methylated PGE2s act through the liver whereas PGE2 may operate through a renal pathway for its response. PGF2 alpha reveals no effect on erythropoietic activity and is no different than that observed for vehicle-treated controls. The prostaglandins tested appear to act primarily through the kidney or liver but the possibility exists that some yet undetermined organ site may also be involved.

The perfusion of livers of partially hepatectomized (H mean) rats lends support to earlier findings that the regenerating rat liver is the source of an erythropoietin-inducing hepatic factor (Ep-IHF) which stimulates hepatic production of extrarenal erythropoietin (Ep). Blood plasma was collected from perfused livers of rats that were partially hepatectomized (H mean) 48 to 72 hrs prior to perfusion with whole blood from normal rats. This plasma, when injected into normal rats which were nephrectomized (N mean) and rendered hypoxic 18 hrs after injection, evoked a significant increase in Ep values when compared to blood plasma collected from perfused livers of normal rats. Ep values were significantly higher when regenerating livers were perfused with blood collected from nephrectomized (N mean) rats than when such livers were perfused with blood of normal rats. The highest Ep values resulting from the liver perfusions were obtained when the liver donor and blood donor rats were both H mean and N mean. The results demonstrate that the liver is the principal source of an Ep-inducing factor since perfusion of the liver eliminated other potential tissue sources of activity in the rat. This was achieved by perfusing the livers directly through the portal vein and collecting the perfusate from the hepatic vein, thereby eliminating potential contributions from organs draining into other parts of the systemic circulation. In addition, it was shown that the kidney inhibits the activity and/or production of the Ep-IHF which is evoked by H mean.

Double partial hepatectomy (hepx) evokes an elevation in serum erythropoietin (Ep) levels in anephric hypoxic animals when compared to non-hypoxic or sham hepx controls. But this Ep response is significantly lower than that found in singly hepx, anephric hypoxic rats. Double hepx also induces numerous cytological changes in the liver. Extravascular accumulation of fat, fibrous scarring, localized necroses, and multiple abscesses, as well as decreased vascularity, occur following the second hepx. A humoral factor was detected in the serum of these animals that is capable of inducing hepatic Ep production when injected into normal rats 18 hours before nephrectomy and hypoxia. This factor, termed hepatopoietin (Hp), was previously demonstrated in the venous serum of singly hepx rats. The serum from animals subjected to double partial hepx is not as potent in inducing Ep production as the serum from singly hepx animals. The discrepancies noted between the single and double hepx groups is attributed to the necrotic cytological changes described above.

Erythropoiesis, which is primarily hepatic in the rat during fetal and early neonatal life, shifts almost entirely to the bone marrow in the neonatal-adolescent stage of development. In the adult, extramedullary erythropoiesis has been demonstrated in the liver and spleen under certain pathological conditions when bone marrow red cell production is insufficient. In the present study, erythropoietic foci have been found in young-adult rat liver regenerating 24-72 hr after subtotal hepatectomy. This erythropoiesis is both extravascular and sinusoidal, with some erythroblastic islands noted. The centrolobular hepatic area contains the highest concentration of erythroblasts. Peripheral blood reticulocytosis coincides with the appearance of these cells and this is considered as an indicator of effective erythropoiesis. Liver regenerating after partial hepatectomy produces significant quantities of erythropoietin (Ep) in response to hypoxia. Subtotal hepatectomy may confer upon the adult liver the ability to revert to a fetal-like condition both in its ability to produce Ep and to function as a hematopoietic inductive microenvironment for erythropoiesis.

Evidence is presented for the existence of a factor in renal venous blood, evoked by subtotal hepatectomy (hepx), which inhibits the production of Ep in nephrectomized (nephrx) rats exposed to hypoxia. Significant inhibitory activity is not observed in blood obtained from sites other than the renal vein. This inhibitory effect is believed to be due to the action of a specific renal inhibitory factor (RIF) which reduces the extrarenal (hepatic) Ep response to hypoxia indirectly, by decreasing the production or effectiveness of an antagonistic liver principle, the hepatic erythropoietic factor (HEF). The HEF has previously been shown to augment hepatic Ep production following hypoxia in renally-deficient animals. The RIF has no anti-Ep action and its activity is not influenced by the accumulation of metabolic wastes. A mechanism for a renal-hepatic antagonism in the Ep response to hypoxia is hypothesized.

Erythropoietin (Ep) is produced mainly by the liver and spleen during fetal and neonatal periods and by the kidney during adolescent and adult life. The liver is also an important extrarenal producer of Ep in the hypoxic, anephric adult animal. Subtotal hepatectomy results in a substantial elevation in serum Ep levels at 30-72 hours after hepatectomy in rats subsequently nephrectomized and rendered hypoxic. Ep production is related to the mass of regenerating liver with peak Ep production occurring during times of greatest tissue proliferation. Regenerative and erythropoietic responses to hepatectomy decline with advancing age. Rats undergoing repeated hepatectomies do not recover full liver mass but the initial rate of regeneration increases following each successive hepatectomy. Ep levels decline in anephric hypoxic rats undergoing multiple hepatectomies when compared to sham-operated controls.

The influence of the pancreas on renal and extrarenal erythropoietin (Ep) production and on the elaboration of the hepatic erythropoietic factor (HEF) was studied in these experiments. Insulin was found to elevate Ep levels in the anephric hypoxic rat when compared to controls, whereas glucagon treatment augmented the hepatic Ep response to hypoxia in the subtotally hepatectomized (hepx) animal while lowering it in the renal intact rat. Production of experimental diabetes either through chemical induction by alloxan or following pancreatectomy diminished the Ep response in all groups tested. Treatment with antiglucagon caused an elevation in the Ep response to hypoxia in the intact rat but lowered Ep levels in the hepx animal. In addition, glucagon and a synthetic hepatotrophic agent (L-histidyl L-lysine acetate) stimulated HEF production in the hepx rat, although none of the agents tested were capable of enhancing HEF levels in the intact rat.

Rats were rendered anaemic by a single bleeding or by a single injection of phenylhydrazine. At various times after the onset of anaemia they were nephrectomized and challenged with a 6 h exposure to hypoxia. The erythropoietin titre observed at the end of this hypoxic period was corrected for renal erythropoietin induced by the anaemia alone, and the resulting extrarenal component was compared to total erythropoietin production of nephric rats in response to anaemia plus 6 h hypoxia. Extrarenal erythropoietin production was found to increase from 10.3% in normal rats to 12.5% in moderately anaemic rats to 15.1% in rats with severe bleeding anaemia. In phenylhydrazine-treated rats this extrarenal component was found to be 18.3% possibly due to stimulation of extrarenal erythropoietin by haemolysed red cells. Chronic phenylhydrazine administration resulted in splenomegaly and Kupffer cell hyperactivity but not in any further stimulation of extrarenal erythropoietin production.

Erythropoietin (Ep) is a glycoprotein hormone that is responsible for mammalian red blood cell production. Adult rat liver regenerating 48-72 h after hepatectomy (hepx) produces elevated levels of Ep in response to hypoxia when compared to sham-operated, anephric hypoxic controls. A factor, termed hepatopoietin (Hp), found in the serum of hepx rats, is capable of stimulating hepatic Ep production when administered to normal rats 18 h prior to hypoxic exposure. Although the hepatic vein is the most potent source of this factor, Hp can also be demonstrated in the systemic arterial circulation. Bilateral nephrectomy (nephrx) of the donor hepx animal 24 h prior to bleeding abolishes this variation, and highest Ep levels are noted when serum from a hepx and nephrx rat is administered to animals immediatley after nephrx and 18 h before hypoxic exposure. Serum derived from hepx male rats displays a greater ability to evoke hepatic Ep production in normal recipients than serum from similarly treated female rats. Regardless of the sex of the hepx donor, Ep elaboration after hypoxia is highest in male recipients. The results indicate that there is a sexual variation in the production of Hp as well as Ep.

Erythropoietin (Ep) in large amounts was detected in extracts of renal tissue from hypoxic rats. These extractions were performed by homogenization of kidney tissue in phosphate buffered saline, centrifugation at 3,000 g and collection of the supernate. Male kidneys contained more Ep than did females and the major portion of Ep is located in the renal cortex. Comparison of intrarenal and plasma Ep levels at various times following initiation and cessation of hypoxia appears to be a useful method for studying the kinetics of erythropoietin production and release, and also for studying feedback mechanisms that influence these functions.

Evidence is presented for production, by the subtotally hepatectomized (hepx) rat, of a factor which induces morphological and physiological hyperactivity of the Kupffer (K) cells and increased formation of Ep by the regenerating liver. This factor, originally termed hepatopoietin (Hp), and more recently the hepatic erythropoietic factor (HEF), is detectable in higher concentration in the hepatic venous blood than in blood draining other organs, thus supporting its hepatic origin. Production of the HEF is best related to hyperactivity of the K cells and not to the parenchymal (P) cells. The HEF can be demonstrated by administering serum from hepx donors to normal rats which respond with increased production of Ep when nephrectomized (nephrx) and rendered hypoxic. Removal of the kidneys from hepx donors further augments the Ep response to this serum in recipients. Subtotal hepx also evokes the production of a renal inhibitory factor (RIF) which reduces the ability of the liver to function as an extrarenal source of Ep. This inhibitor is found in renal venous blood and not in blood draining other organs. It is suggested that the RIF reduces the hepatic Ep response to hypoxia by diminishing the production and/or activity of the HEF. The RIF possesses no anti-Ep activity and its appearance and actions are not influenced by accumulation of metabolic wastes (as in the nephrx or ureterally-ligated rat). Erythroblastic nests have been observed in regenerating livers at 24-48 hr after subtotal hepx. It would seem that removal of a considerable part of the liver, which stimulates hepatic regeneration, confers upon this organ an increased ability to produce Ep and to function as a hematopoietic inductive microenvironment (HIM) for erythropoiesis.

Erythropoietin production in response to hypoxic-hypoxia is markedly reduced in the newborn when compared to the adult rat. This response improves steadily with age and reaches adult values at about 4 wk. When animals of the same age are stimulated with anemic-hypoxia, considerably higher levels of erythropoietin are found. The erythropoietin level is proportional to the degree of anemia and independent of the age of the animal. Extraction of erythropoietin from tissue homogenates revealed a parallelism between the plasma and kidney erythropoietin content, while no erythropoietin could be extracted from liver tissue at any age. The lack of response to hypoxia in the newborn appears to be related to the high hemoglobin oxygen affinity during the neonatal period, which facilitates oxygen loading. Newborn rats have a very low intraerythrocytic concentration of 2-3 DPG and a marked shift to the left in the oxygen hemoglobin dissociation curve that slowly increases to adult values at 4 wk of age. The response to anemia on the other hand, appears to be normal and not affected by age or by hemoglobin oxygen affinity. These studies suggest that the newborn rat, when properly stimulated, is able to produce normal amounts of erythropoietin, most likely renal in origin.

Experiments were performed to determine the erythropoietin (Ep) content of homogenates of kidneys and livers of male and female rats of various ages. In all studies, homogenates were adjusted to a concentration of 4 g of tissue per 12 ml of phosphate-buffered-saline, and the stimulus to Ep production consisted of exposure to 0.42 atmosphere for 4 h. The concentration of Ep in kidneys of male rats was about three times that found in those of females and was contained predominantly in the cortical portion of the kidneys. Ep was not detectable in kidneys of rats younger than 3 weeks of age, and reached a maximum concentration after 4 weeks of age. The Ep content of the liver was barely detectable regardless of the age of the rat or its plasma Ep titer; and did not increase significantly by administering angiotensin II or CCl4 (substances which increase extrarenal Ep production).

Plasma titers of erythropoietin (Ep) are known to increase initially during hypoxia and to return then towards prehypoxia values. To find out if this pattern of plasma Ep might be related to changes in the production of the hormone, I have compared plasma with kidney Ep titers in hypoxic rats. Rats were exposed to hypoxia in a hypobaric chamber at 0.42 atm for various time intervals for up to 4 days. Kidney Ep titers were assayed in extracts from kidneys that had been flushed free of blood in situ. It was found that kidneys of normal rats do not store significant amounts of Ep. Kidney Ep titers increased transiently during hypoxia. They reached maximum values after 6h and then declined to almost undetectable levels at continued hypoxia. In the plasma, maximum values were found after 12-18h of hypoxia. Additional studies were done on the effects of discontinuous hypoxia. It was found that, even after 3 days of previous hypoxia exposure, plasma and kidney Ep titers increased again in rats when these were maintained intermittently in normoxia for 18 h. It is concluded that the rise and fall in plasma Ep titers during hypoxia reflect similar changes in kidney Ep production.

The effects of infusing subpressor doses of angiotensin II into hypoxic and anemic rats on plasma Ep levels were determined. The effect was greatest when 5 micrograms of angiotensin II per hour was infused into rats made hypoxic 18 hr after nephrectomy. Infusion of larger amounts of angiotensin II had a lesser effect on extrarenal Ep production than did infusion of 5 micrograms/hr. Infusion of angiotensin II into rats nephrectomized 1 hr prior to exposure to hypoxia affected extrarenal Ep production to a lesser degree than the infusion into rats nephrectomized 18 hr prior to hypoxia. In contrast, administration of carbon tetrachloride per os stimulated extrarenal Ep production only when nephrectomy was performed just prior to exposure to hypoxia. Administration of both CCl4 and angiotensin II to hypoxic anephric rats elevated the plasma Ep level to approximately 1.0 IRP U/ml.

Evidence is presented for the existence of a factor in renal venous blood, evoked by subtotal hepatectomy (hepx), which inhibits the production of Ep in nephrectomized (nephrx) rats exposed to hypoxia. Significant inhibitory activity is not observed in blood obtained from sites other than the renal vein. This inhibitory effect is believed to be due to the action of a specific renal inhibitory factor (RIF) which reduces the extrarenal (hepatic) Ep response to hypoxia indirectly, by decreasing the production or effectiveness of an antagonistic liver principle, the hepatic erythropoietic factor (HEF). The HEF has previously been shown to augment hepatic Ep production following hypoxia in renally-deficient animals. The RIF has no anti-Ep action and its activity is not influenced by the accumulation of metabolic wastes. A mechanism for a renal-hepatic antagonism in the Ep response to hypoxia is hypothesized.