Linked Life Data

9242564

Source:http://linkedlifedata.com/resource/pubmed/id/9242564

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1997-8-25
pubmed:abstractText
Using homologous recombination, both EKLF alleles in murine embryonic stem (ES) cells were inactivated. These EKLF-/- ES cells were capable of undergoing in vitro differentiation to form definitive erythroid colonies that were similar in size and number to those formed by wild-type ES cells. However, the EKLF-/- colonies were poorly hemoglobinized and enucleated erythrocytes in these colonies contained numerous Heinz bodies. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses revealed that adult and embryonic globin genes were appropriately regulated, with the exception of beta h1-globin, which continued to be expressed at a very low level. The ratio of adult beta-globin/alpha-globin mRNA in the mutant ES cells was 1/15 of that in wild-type ES cells. When the EKLF-/- cells were injected into blastocysts, they did not contribute at a detectable level to the mature erythrocyte compartment of the chimeric animals, based on analysis of glucose phosphate isomerase-1 (GPI-1) isozymes and hemoglobins that distinguish ES cell-derived erythrocytes from host blastocyst-derived erythrocytes. In contrast, semiquantitative RT-PCR analysis of RNA from reticulocytes of the same chimeric animals suggested that the ES cell-derived reticulocytes were present at a level of 6% to 8%. This indicated that the EKLF-/- erythrocytes in adult animals must be short-lived, apparently due to the imbalance of beta-versus alpha-globin chains, leading to the precipitation of excess alpha-globin chains to form Heinz bodies. Consistent with this hypothesis, the short life span was ameliorated by introduction into the EKLF-/- ES cells of a human LCR/gamma-globin gene, as evidenced by the presence of ES cell-derived reticulocytes as well as mature erythrocytes in the blood of the chimeric animals.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
AIM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0006-4971
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
90
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
1291-9
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:9242564-Animals, pubmed-meshheading:9242564-Cell Differentiation, pubmed-meshheading:9242564-Chimera, pubmed-meshheading:9242564-DNA-Binding Proteins, pubmed-meshheading:9242564-Erythrocyte Aging, pubmed-meshheading:9242564-Erythroid Precursor Cells, pubmed-meshheading:9242564-Erythropoiesis, pubmed-meshheading:9242564-Gene Expression Regulation, Developmental, pubmed-meshheading:9242564-Gene Targeting, pubmed-meshheading:9242564-Gene Therapy, pubmed-meshheading:9242564-Genes, Switch, pubmed-meshheading:9242564-Genes, Synthetic, pubmed-meshheading:9242564-Globins, pubmed-meshheading:9242564-Humans, pubmed-meshheading:9242564-Kruppel-Like Transcription Factors, pubmed-meshheading:9242564-Liver, pubmed-meshheading:9242564-Mice, pubmed-meshheading:9242564-Mice, Knockout, pubmed-meshheading:9242564-Polymerase Chain Reaction, pubmed-meshheading:9242564-Recombinant Fusion Proteins, pubmed-meshheading:9242564-Regulatory Sequences, Nucleic Acid, pubmed-meshheading:9242564-Reticulocytes, pubmed-meshheading:9242564-Species Specificity, pubmed-meshheading:9242564-Transcription, Genetic, pubmed-meshheading:9242564-Transcription Factors, pubmed-meshheading:9242564-beta-Thalassemia
pubmed:year
1997
pubmed:articleTitle
A shortened life span of EKLF-/- adult erythrocytes, due to a deficiency of beta-globin chains, is ameliorated by human gamma-globin chains.
pubmed:affiliation
Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't