Comparative studies of erythroid growth factors

Representative specimens from two classes of Vertebrata Sub-Phyllum, Bufo paracnemis (amphibian) and Gallus domesticus (avian) were made anemic by phenylhydrazine treatment. Appearance of serum factors able to stimulate the proliferation of mammalian erythroid cells was tested. Normal and anemic sera from Gallus domesticus and Bufo paracnemis were fractioned by alcoholic treatment and assayed by the post-hypoxic mice method, showing null uptake of 59Fe. When assayed in semisolid cultures using bone marrow murine cells at different times of incubation (CFU-E and BFU-E colonies), anemia Gallus domesticus serum showed high stimulatory activity, while anemic Bufo paracnemis serum was unable to enhance erythroid proliferation. Gel filtration chromatography of partially purified avian samples on Sephadex G-150 showed three molecular entities responsible for biological activity in vitro, with an apparent molecular weight of 29, 14 and 10 KD respectively. They were submitted to several treatments and then tested for biological activity. All factors were heat stable, sensitive to neuraminidase treatment, while dithiothreitol caused loss of activity on low molecular weight proteins. These results suggest at least under these experimental conditions, the presence of analogous erythroid factors among homeotherms amniotas.