Juaristi J A, Aguirre M V, Guillén E R, Alvarez M A, Brandan N C
Cátedra de Bioquímica, Instituto de Investigaciones Biofarmacológicas, Facultad de Medicina, U.N. del Nordeste, Corrientes, R. Argentina.
Acta Physiol Pharmacol Latinoam. 1989;39(2):133-44.
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.
选取了脊椎动物亚门两个纲的代表性样本,即南美蟾蜍(两栖类)和家鸡(鸟类),通过苯肼处理使其贫血。测试了能够刺激哺乳动物红细胞系细胞增殖的血清因子的出现情况。对家鸡和南美蟾蜍的正常血清及贫血血清进行乙醇处理分级,并采用低氧后小鼠法进行检测,结果显示对59Fe无摄取。在使用骨髓小鼠细胞进行不同孵育时间的半固体培养(CFU-E和BFU-E集落)中进行检测时,家鸡贫血血清显示出高刺激活性,而南美蟾蜍贫血血清则无法增强红细胞增殖。在Sephadex G - 150上对部分纯化的鸟类样本进行凝胶过滤层析,显示有三种分子实体负责体外生物活性,其表观分子量分别为29、14和10千道尔顿。对它们进行了几种处理,然后测试其生物活性。所有因子均耐热,对神经氨酸酶处理敏感,而二硫苏糖醇会导致低分子量蛋白失去活性。这些结果表明,至少在这些实验条件下,恒温羊膜动物中存在类似的红细胞生成因子。