Isolation of an erythropoietic stimulating factor from the serum of anemic chicks

A factor stimulating fowl erythropoiesis in vitro has been isolated from the blood of anemic chick by gel filtration and ion exchange chromatography. This purification procedure yields a preparation whose specific activity is enriched 44 times and which has been relieved of toxic factors present in the crude anemic serum. In vitro erythropoiesis requires addition into the culture medium of erythropoietic stimulating factor and of another factor present in the serum of anemic as well as non-anemic chick. This last factor is lost during purification of the erythropoietic stimulating factor and is not present in fetal calf serum.     

Recovery of erythropoietin from anemic sheep plasma

A method is described for the large-scale preparation of erythropoietin from anemic sheep plasma. DEAE-cellulose and carboxymethylcellulose column chromatography was used to prepare Step II erythropoietin. A total of 168 sheep yielded 499 liters of plasma from which 323,000 IU of Step II erythropoietin was obtained.     

Studies of the development of frog hemopoietic tissue in vitro. I. Spleen culture assay of an erythropoietic factor in anemic frog blood.

A new in vitro technique has been described for demonstrating the presence of an erythropoietic factor in the circulating blood of frogs. The assay system consisted of MC33 medium, erythropoietically active spleen cells from Rana pipiens, and plasma or serum from frogs made anemic via phenylhydrazine or bleeding. The spleen cells, which remain erythropoietically active for up to nine days, were found to incorporate 59Fe, [3H]thymidine, [3H]uridine, and [3H]leucine at a greater rate in the presence of plasma or serum from anemic versus normal frogs. The hormones triiodothyronine, prolactin, and erythropoietin were not effective in eliciting an hemopoietic response. The data presented suggest that the spleen from that adult frog is a major site of erythroid differentiation and maturation.     

Integrin participates in the effect of thyroxine on plasma membrane in immature rat testis

Background

The secretory activity of Sertoli cells (SC) is dependent on ion channel functions and protein synthesis and is critical to ongoing spermatogenesis. The aim of this study was to investigate the mechanism of action associated with a non-metabolizable amino acid [¹⁴C]-MeAIB (α-(methyl-amino)isobutyric acid) accumulation stimulated by T₄ and the role of the integrin receptor in this event, and also to clarify whether the T₄ effect on MeAIB accumulation and on Ca²⁺ influx culminates in cell secretion.

Methods

We have studied the rapid and plasma membrane initiated effects of T₄ by using ⁴⁵Ca²⁺ uptake and [⁴⁵C]-MeAIB accumulation assays, respectively. Thymidine incorporation into DNA was used to monitor nuclear activity and quinacrine to analyze the secretory activity on SC.

Results

The stimulation of MeAIB accumulation by T₄ appears to be mediated by the integrin receptor in the plasma membrane since tetrac and RGD peptide were able to nullify the effect of this hormone. In addition, T₄ increases extracellular Ca²⁺ uptake and Ca²⁺ from intracellular stocks to enhance nuclear activity, but this genomic action seems not to influence SC secretion mediated by T₄. Also, the cytoskeleton and ClC-3 chloride channel contribute to the membrane-associated responses of SC.

Conclusions

T₄ integrin receptor activation ultimately determines the plasma membrane responses on amino acid transport in SC, but it is not involved in calcium influx, cell secretion or the nuclear effect of the hormone.

General significance

The integrin receptor activation by T₄ may take a role in plasma membrane processes involved in the male reproductive system.     

Studies on the erythropoietic cell system in CBA mice after Rauscher virus infection.

Erythropoiesis in CBA mice was studied in Rauscher leukemia virus infected mice using the incorporation of 59Fe into spleen, bone marrow and peripheral blood. Beginning at day 4 an increased uptake into the spleen and a decrease in the bone marrow and the peripheral blood was observed. The increased uptake by the spleen was also found in plethoric mice. The erythropoietin responsive compartment was also enlarged in the spleen of these mice. The The dose-response-curve for erythropoietin was altered 4 days after infection, there was a higher background level of 59Fe incorporation and the response to low doses was better in infected animals. The reticulocytopenia which is usually seen in these mice, was overcome by administration of high doses of erythropoietin. It is concluded that the Rauscher virus acts in a similar manner to erythropoietin, but the erythropoiesis induced is ineffective since the cells do not mature. This maturation deficiency is influenced by administration of exogenous erythropoietin.