Identification of erythroid-enriched gene expression in the mouse embryonic yolk sac using microdissected cells.
Little is known about the genes that control the embryonic erythroid program. Laser capture microdissection was used to isolate primitive erythroid precursors and epithelial cells from frozen sections of the embryonic day 9.5 yolk sac. The RNA samples were amplified and labeled for hybridization to Affymetrix GeneChip Mouse Genome 430A 2.0 arrays. Ninety-one genes are expressed significantly higher in erythroid than in epithelial cells. Ingenuity pathway analysis indicates that many of these erythroid-enriched genes cluster in highly significant biological networks. One of these networks contains RBTN2/LMO2, SCL/TAL1, and EKLF/KLF1, three of the very few genes required for primitive erythropoiesis. Quantitative real-time polymerase chain reaction was used to verify that platelet factor 4, reelin, thrombospondin-1, and muscleblind-like 1 mRNA is erythroid-enriched. These genes have established roles in development or differentiation in other systems, and are, therefore, good candidates for regulating primitive erythropoiesis. These results provide a catalog of genes expressed during primitive erythropoiesis.
Redmond
Latasha C
LC
Department of Human Genetics, Virginia Commonwealth University, Richmond, Virginia 23298-0035, USA.
Dumur
Catherine I
CI
Archer
Kellie J
KJ
Haar
Jack L
JL
Lloyd
Joyce A
JA
D000818:Animals; D004847:Epithelial Cells / Q000378:metabolism; D041905:Erythroid Cells / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D053263:Gene Regulatory Networks / Q000235:genetics*; D051379:Mice; D042282:Microdissection; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000235:genetics; D000091042:Reelin Protein; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015017:Yolk Sac / Q000196:embryology / Q000378:metabolism*
D009841:Oligonucleotides; D000091042:Reelin Protein; C000717127:RELN protein, human; C000717128:Reln protein, mouse