In-vivo inhibition of Fgfr2b signalling in FACS-isolated Sftpc-expressing cells in embryonic mouse lungs from E14.5 to E18.5

ABSTRACT: We are interested in studying the role of FGF/FGFR2b signalling on lung alveolar epithelial bipotent progenitors. Bipotent progenitors give rise to alveolar type 1 and type 2 cells (AECI and AECII, respectively). Previous research has shown that AECII cells isolated from lungs where FGF signalling has been globally inhibited show a gene signature much closer to type 1 cells. Furthermore, the relative number of AECI in experimental lungs was increased compared to control lungs. This suggests FGF signalling promotes AECII differentiation. We want to test the hypothesis that FGF signalling promotes the differentiation of AECII by signalling to bipotent progenitors. To test this hypothesis, we have used a transgenic mouse line (SpcCreERT2_Fgfr2bflox_tomatoflox), which, upon administration of tamoxifen, can be used to knock-out the FGFR2b receptor in SFTPC + cells, while labelling those cells red. We injected mice carrying embryos with and without the Fgfr2bflox allele (experimental and control, respectively) at E14.5 and at E15.5. At this timepoint, SFTPC is a marker for bipotent progenitors. Thus, by administering tamoxifen at this timepoint, bipotent progenitors were targeted to knock-out FGFR2b, and thus inhibit FGF/FGFR2b signalling to these cells. We harvested lungs at E18.5, a timepoint where the differentiation of AECI and AECII is well underway. Harvested lungs were prepared for FACS-based cell sorting. Cells were sorted by gating for total epithelial cells, then RFP + cells (representing bipotent progenitors), then AECI and AECII cells. These cells were collected, and their total RNA isolated for gene array analysis. We will then compare the AECI and AECII gene signatures of isolated cells between experimental and control samples.

ORGANISM(S): Mus musculus

PROVIDER: GSE115879 | GEO | 2021/06/01

REPOSITORIES: GEO