RESUMO
Goat antiserum was prepared against erythrocyte membranes obtained from a man who had X-linked muscular dystrophy. When cross-adsorbed with control membranes and tested on double immunodiffusion plates or by binding to intact erythrocytes suspended in thin-layer agar plates, this antiserum discriminated between membranes from phenotypically normal controls and dystrophic men, and between controls and obligate female dystrophy carriers. It also identified two populations among a small sample of women at risk for being dystrophy carriers.
Assuntos
Antígenos/imunologia , Membrana Eritrocítica/imunologia , Eritrócitos/imunologia , Distrofias Musculares/imunologia , Adolescente , Adulto , Anticorpos/genética , Anticorpos/imunologia , Antígenos/genética , Criança , Membrana Eritrocítica/ultraestrutura , Feminino , Ligação Genética , Humanos , Imunodifusão , Masculino , Pessoa de Meia-Idade , Distrofias Musculares/genética , Distrofias Musculares/patologia , Linhagem , Cromossomo XRESUMO
Inherited electrophoretic variations of hemoglobin, carbonic anhydrase, and glucose-6-phosphate dehydrogenase in individual erythrocytes were separated by electrophoresis in ultrathin agar gels. By staining the electropherograms with specific fluorescein-conjugated antibodies against hemoglobins, relative proportions of two hemoglobins within individual erythrocytes can be estimated. The findings suggested that the intracellular proportions of HbA and HbS in heterozygotes are heterogeneous within a given population of cells. By this method cells containing hemoglobin F (F cells) as well as a minor variant of hemoglobin F were identified. This tool potentially offers an approach to monitoring distribution of inherited variants in individual erythrocytes for a large number of proteins.
Assuntos
Anidrases Carbônicas/genética , Eritrócitos/análise , Variação Genética , Glucosefosfato Desidrogenase/genética , Hemoglobinas/genética , Animais , Eletroforese em Gel de Ágar , Feminino , Imunofluorescência , Humanos , Recém-Nascido , Macaca/genéticaAssuntos
Eletroforese das Proteínas Sanguíneas , Anidrases Carbônicas/sangue , Eletroforese em Acetato de Celulose/instrumentação , Eletroforese/instrumentação , Hemoglobinas/análise , Animais , Eletroforese das Proteínas Sanguíneas/instrumentação , Eletroforese em Acetato de Celulose/métodos , Eritrócitos/enzimologia , Haplorrinos , Humanos , MacacaRESUMO
We present here a method for electrophoretic identification of hemoglobin variants in many individual erythrocytes simultaneously, which is made possible by instruments developed in our laboratory. The system spaces and aligns the erythrocytes along a common origin and prevents 'doubling' of cells. Unambiguous separation of different hemoglobins (Hbs) occurred from single erythrocytes, such as Hbs A, C, F. Staged micrographs showed that hemoglobins of differing mobility from a single cell arise from a single point source of hemoglobin. In principle this capability is applicable to other proteins and could facilitate investigation at a cellular level into a variety of questions pertaining to gene action.
Assuntos
Eletroforese/métodos , Eritrócitos/análise , Hemoglobinas/análise , Eletroforese/instrumentação , Variação Genética , HumanosRESUMO
The development of highly specific fluorescent labelled antibodies against haemoglobin F presented an opportunity to investigate variables which might influence distribution of this haemoglobin among individual erythrocytes. Earlier investigations revealed heterogeneous distribution within healthy individuals and in individuals with sickle cell disease and other haemoglobinopathies. The current study demonstrates quantitatively that there are normal biological determinants of variability in the frequency of F-containing erythrocytes in individuals who have a haemoglobin A electrophoretic phenotype. It also is shown that the frequency of F-containing erythrocytes during the early recovery phase of sickle cell crisis is significantly higher than when there is no recent history of crisis. The mean quantity of ahemoglobin F per F-containing erythrocyte appears to be lower after a crisis than in individuals without recent history of crisis. This suggests that following a crisis there may either be biochemical constraints on haemoglobin F synthesis per erythrocyte precursor cell or else there is limited opportunity for selective removal of low F-containing erythrocytes from the circulation.
Assuntos
Eritrócitos/análise , Hemoglobina Fetal/análise , Variação Genética , Adolescente , Adulto , Anemia Falciforme/sangue , Criança , Feminino , Imunofluorescência , Hemoglobina Falciforme/análise , Humanos , Imunodifusão , Masculino , Fatores de TempoRESUMO
Effects of thyroxine on incorporation of L-serine-C14 into four carbonic anhydrase isozymes (CA II, CA Ia, CA Ib, CA Ic) and hemoglobin were quantified in reticulocytes of Macaca nemestrian in vitro. Response to thyroxine differed significantly between CA Ia and two allelic variants (CA Ib and CA Ic) and the nonallelic isozyme (CA II). The effects of thyroxine on serine incorporation into hemoglobin and three of the carbonic anhydrase isozymes were shown to be nonlinear with thyroxine concentration.