Elsevier

Gene Expression Patterns

Volume 12, Issues 3–4, March–April 2012, Pages 154-163
Gene Expression Patterns

Novel cell surface genes expressed in the stomach primordium during gastrointestinal morphogenesis of mouse embryos

https://doi.org/10.1016/j.gep.2012.01.001Get rights and content

Abstract

The mechanisms of gastrointestinal morphogenesis in mammals are not well understood. This is partly due to the lack of appropriate markers that are expressed with spatiotemporal specificity in the gastrointestinal tract during development. Using mouse embryos, we surveyed markers of the prospective stomach region during gastrointestinal morphogenesis. The initiation of organ bud formation occurs at E10.5 in mice. These primordia for the digestive organs protrude from a tube-like structured endoderm and have their own distinct morphogenesis. We identified 3 cell surface genes – Adra2a, Fzd5, and Trpv6 – that are expressed in the developing stomach region during gastrointestinal morphogenesis using a microarray-based screening. These novel genes will be useful in expanding our understanding of the mechanisms of gastrointestinal development.

Highlights

► 3 Novel cell surface genes expressed in the stomach primordium were identified. ► Adra2a is expressed in the mesenchyme of the mouse stomach primordium at E11.5. ► Fzd5 and Trpv6 are expressed in the epithelial layer of the stomach primordium after E11.5.

Section snippets

Results and discussion

During mouse gastrulation, development of the embryonic endoderm is initiated by an evolutionally conserved gene regulatory network. Nodal, a member of the transforming growth factor-β superfamily, is one of the major growth factors that promote endoderm development. High levels of nodal expression, maintained by a positive autoregulatory loop, induce the conserved network of transcription factors for endoderm development (Shen, 2007). The resulting endodermal cells then morphologically

Embryo collection

Mouse embryos were collected from pregnant ICR mice (E9.5–E17.5) in ice-cold PBS. The embryos were fixed in 4% paraformaldehyde in PBS solution overnight at 4 °C with gentle shaking. The embryos were washed in PBS for 10 min at 4 °C and dehydrated using a series of methanol-PBS washes. The embryos were then stored in 100% methanol at −20 °C until in situ hybridization analysis. Newborn mice were also fixed after being anesthetized. All animal experiments were conducted according to the committee

Acknowledgments

This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (M.A. and A.K.) and a research grant from the Suzuken Memorial Foundation (A.K.).

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      Trpv5, on the contrary, only has reported expression data at E11.5 and E15.5 in mouse (Magdaleno et al., 2006). Data on trpv6 embryonic expression pattern is available only after E9.5 and at 3dpf in zebrafish (Magdaleno et al., 2006; Diez-Roux et al., 2011; Naguchi et al., 2012). Given the importance and versatility of trpv channels, a more comprehensive characterization during development will provide important insight into some of their earlier functions.

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