title: Klf5 regulates lineage formation in the pre-implantation mouse embryo.
abstract: Kruppel-like transcription factors (Klfs) are essential for the induction and maintenance of pluripotency of embryonic stem cells (ESCs), yet little is known about their roles in establishing the three lineages of the pre-implantation embryo. Here, we show that Klf5 is required for the formation of the trophectoderm (TE) and the inner cell mass (ICM), and for repressing primitive endoderm (PE) development. Although cell polarity appeared normal, Klf5 mutant embryos arrested at the blastocyst stage and failed to hatch due to defective TE development. Klf5 acted cell-autonomously in the TE, downstream of Fgf4 and upstream of Cdx2, Eomes and Krt8. In the ICM, loss of Klf5 resulted in reduced expression of pluripotency markers Oct4 and Nanog, but led to increased Sox17 expression in the PE, suggesting that Klf5 suppresses the PE lineage. Consistent with this, overexpression of Klf5 in transgenic embryos was sufficient to suppress the Sox17(+) PE lineage in the ICM. Klf5 overexpression led to a dose-dependent decrease in Sox17 promoter activity in reporter assays in cultured cells. Moreover, in chimeric embryos, Klf5(-/-) cells preferentially contributed to the Sox17(+) PE lineage and Cdx2 expression was not rescued in Klf5(-/-) outer cells. Finally, outgrowths from Klf5(-/-) embryos failed to form an ICM/pluripotent colony, had very few Oct4(+) or Cdx2(+) cells, but showed an increase in the percentage of Sox17(+) PE cells. These findings demonstrate that Klf5 is a dynamic regulator of all three lineages in the pre-implantation embryo by promoting the TE and epiblast lineages while suppressing the PE lineage.
authors.affiliation: Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229-3039, USA.
authors.forename: Suh-Chin J
authors.initials: SC
authors.lastname: Lin
authors.forename: Maqsood A
authors.initials: MA
authors.lastname: Wani
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Whitsett
authors.forename: James M
authors.initials: JM
authors.lastname: Wells
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism*; D000071616:CDX2 Transcription Factor; D002452:Cell Count; D016923:Cell Death; D019070:Cell Lineage; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D003433:Crosses, Genetic; D004475:Ectoderm / Q000166:cytology / Q000378:metabolism; D004707:Endoderm / Q000166:cytology / Q000378:metabolism; D005260:Female; D051519:Fibroblast Growth Factor 4 / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D005838:Genotype; D005855:Germ Layers / Q000166:cytology / Q000378:metabolism; D024002:HMGB Proteins / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D051741:Kruppel-Like Transcription Factors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D008297:Male; D051379:Mice; D055760:SOXF Transcription Factors / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D015415:Biomarkers; D000071616:CDX2 Transcription Factor; C487514:Cdx2 protein, mouse; C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; D024002:HMGB Proteins; D018398:Homeodomain Proteins; C118391:Klf5 protein, mouse; D051741:Kruppel-Like Transcription Factors; D055760:SOXF Transcription Factors; C511606:Sox17 protein, mouse; D014157:Transcription Factors