High Performance Liquid Chromatography of Rat Globin Chains of Fetal Liver During the Switch from Embryonic to Adult Hemoglobin

Abstract

A reversed-phase high performance liquid chromatography (HPLC) method for the direct separation of globin chains from cell lysates of peripheral blood and fetal liver from rat fetuses is described. Partial amino acid analysis of the globins eluted from the HPLC columns as well as comparison with the known elution positions of the adult globin chains in carboxymethyl cellulose chromatography indicated that α-chains are eluted first, followed by adult β chains. The last chains to be eluted are considered as embryonic globins because of their absence in adult rats and their rapid disappearance from the liver after the 14th day of gestation. Liver erythroid cells isolated from rat fetuses on day 14 of gestation mainly synthesized α-chains and embryonic globin chains, whereas cells prepared from 16-day old fetuses synthesized almost exclusively α-chains and adult β chains. When the fetal rat liver cells were cultured for 20h with erythropoietin there was a significant stimulation in the synthesis of α-globins and adult β chains but not on the synthesis of embryonic globin chains. It is concluded that HPLC can be useful for the study of rat globin chain synthesis during fetal development, because it separates the globin chains in the three groups of globins, namely alpha, beta and embryonic chains which are important in the switch occurring in the liver.