HbF synthesis during stress erythropoiesis as determined by gamma-mRNA/non-alpha-mRNA quantification.
To determine whether a quantitative relationship exists between globin mRNAs and their translation product during stress erythropoiesis in infants with increased production of fetal hemoglobin (HbF), we measured and compared the relative amounts of the mRNAs of alpha-, beta-, and gamma-globins and their protein synthesis. The synthesis of globin in immature red cells was determined by the incorporation of [3H]leucine, followed by separation and quantification of the polypeptides by C4-reverse phase HPLC. The relative proportions of the mRNAs of the globins were determined by RNase protection assay. A comparison of blood samples from 17 infants expected to have increased production of HbF in relation to their developmental age (five infants of diabetic mothers, two infants with intrauterine growth retardation, one infant with bronchopulmonary dysplasia, and seven infants with cyanotic heart disease) revealed a very significant correlation (r2 = 0.994; p < 0.001) between the ratio of globin mRNAs encoding HbF ([gamma/(gamma + beta)] mRNAs) and the ratios of the de novo synthesis of HbF [gamma/(gamma + beta)]. When only the 10 infants who had increased HbF synthesis are included, the correlation remains unchanged (r2 = 0.997, p < 0.001). The data demonstrated that under conditions of erythropoietic stress, when HbF production is increased, there is a close relationship between the quantification of gamma-globin mRNA and gamma-globin synthesis. The usual methods of determining HbF synthesis can be replaced by globin mRNA determination, which can be performed rapidly with a minimal amount of blood.
Bard
H
H
Department of Pediatrics, University of Montreal, Hospital Sainte-Justine, Canada. bardh@ere.umontreal.ca
Gagnon
C
C
Peri
K G
KG
D000375:Aging; D001997:Bronchopulmonary Dysplasia / Q000097:blood*; D003490:Cyanosis; D004920:Erythropoiesis; D005260:Female; D005312:Fetal Blood; D005317:Fetal Growth Retardation / Q000097:blood*; D005319:Fetal Hemoglobin / Q000235:genetics*; D005914:Globins / Q000235:genetics*; D006330:Heart Defects, Congenital / Q000097:blood*; D006801:Humans; D007231:Infant, Newborn / Q000097:blood*; D011247:Pregnancy; D011254:Pregnancy in Diabetics; D012333:RNA, Messenger / Q000032:analysis*; D012016:Reference Values; D012044:Regression Analysis; D013312:Stress, Physiological; D014158:Transcription, Genetic
D012333:RNA, Messenger; D005914:Globins; D005319:Fetal Hemoglobin