title: Endoderm complexity in the mouse gastrula is revealed through the expression of spink3.
abstract: Endoderm formation in the mammalian embryo occurs first in the blastocyst, when the primitive endoderm and pluripotent cells resolve into separate lineages, and again during gastrulation, when the definitive endoderm progenitor population emerges from the primitive streak. The formation of the definitive endoderm can be modeled using pluripotent cell differentiation in culture. The differentiation of early primitive ectoderm-like (EPL) cells, a pluripotent cell population formed from embryonic stem (ES) cells, was used to identify and characterize definitive endoderm formation. Expression of serine peptidase inhibitor, Kazal type 3 (Spink3) was detected in EPL cell-derived endoderm, and in a band of endoderm immediately distal to the embryonic-extra-embryonic boundary in pregastrula and gastrulating embryos. Later expression marked a region of endoderm separating the yolk sac from the developing gut. In the embryo, Spink3 expression marked a region of endoderm comprising the distal visceral endoderm, as determined by an endocytosis assay, and the proximal region of the definitive endoderm. This region was distinct from the more distal definitive endoderm population, marked by thyrotropin-releasing hormone (Trh). Endoderm expressing either Spink3 or Trh could be formed during EPL cell differentiation, and the prevalence of these populations could be influenced by culture medium and growth factor addition. Moreover, further differentiation suggested that the potential of these populations differed. These approaches have revealed an unexpected complexity in the definitive endoderm lineage, a complexity that will need to be accommodated in differentiation protocols to ensure the formation of the appropriate definitive endoderm progenitor in the future.
authors.affiliation: Department of Zoology, University of Melbourne , Victoria, Australia .
authors.forename: Hwee Ngee
authors.initials: HN
authors.lastname: Goh
authors.affiliation: The Menzies Research Institute Tasmania, University of Tasmania , Tasmania, Australia .
authors.forename: Peter D
authors.initials: PD
authors.lastname: Rathjen
authors.affiliation: Department of Zoology, University of Melbourne , Victoria, Australia .
authors.forename: Mary
authors.initials: M
authors.lastname: Familari
authors.affiliation: Department of Zoology, University of Melbourne , Victoria, Australia . ; The Menzies Research Institute Tasmania, University of Tasmania , Tasmania, Australia .
authors.forename: Joy
authors.initials: J
authors.lastname: Rathjen
keywords: cellular biology; developmental biology; stem cells