Characterization of the mid-foregut transcriptome identifies genes regulated during lung bud induction
Section snippets
Developmental lung genes are expressed in the prospective lung region of the foregut
We studied temporal differences in gene expression in mid-foregut tissues containing endoderm and mesoderm cells isolated by microdissection at three developmental stages prior and after lung bud formation (16–20, 21–25, and 26–30 somite stages) (Fig. 1a and b). We determined by real-time RT-PCR (QRT-PCR) whether the tissues to be used in the microarray studies expressed lung and liver marker genes. Twenty-four hours before lung budding, at E8.5, the mid-foregut already expresses lung
Isolation of mid-foregut tissue
We selected based on morphological features, three developmental stages to study gene expression profiles of mid-foregut tissue, containing endoderm and mesoderm cells. The stages are: 16–20 somite embryos, 21–25 somite embryos and 25–30 somite embryos. The earliest samples (16–20 somites) show evidence of primitive thyroid and liver/pancreas budding which provide morphologic limits at the extremes of the mid-foregut field from which the lung is derived. All embryos with greater than 25 somites
Acknowledgements
We thank Drs. Mary C. Williams, Jerome S. Brody, Wellington V. Cardoso, and YuXia Cao for thoughtful comments about the manuscript. Dr. Norman Gerry for his technical support in the microarray data analysis. This work was supported by NHLBI Program Project Grant HL47049, and Doris Duke Charitable Foundation Clinical Scientist Development Award (A.S.).
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