Characterization of a novel fibroblast growth factor 10 (Fgf10) knock-in mouse line to target mesenchymal progenitors during embryonic development.
Fibroblast growth factor 10 (Fgf10) is a key regulator of diverse organogenetic programs during mouse development, particularly branching morphogenesis. Fgf10-null mice suffer from lung and limb agenesis as well as cecal and colonic atresia and are thus not viable. To date, the Mlcv1v-nLacZ-24 transgenic mouse strain (referred to as Fgf10(LacZ)), which carries a LacZ insertion 114 kb upstream of exon 1 of Fgf10 gene, has been the only strain to allow transient lineage tracing of Fgf10-positive cells. Here, we describe a novel Fgf10(Cre-ERT2) knock-in line (Fgf10(iCre)) in which a Cre-ERT2-IRES-YFP cassette has been introduced in frame with the ATG of exon 1 of Fgf10 gene. Our studies show that Cre-ERT2 insertion disrupts Fgf10 function. However, administration of tamoxifen to Fgf10(iCre); Tomato(flox) double transgenic embryos or adult mice results in specific labeling of Fgf10-positive cells, which can be lineage-traced temporally and spatially. Moreover, we show that the Fgf10(iCre) line can be used for conditional gene inactivation in an inducible fashion during early developmental stages. We also provide evidence that transcription factors located in the first intron of Fgf10 gene are critical for maintaining Fgf10 expression over time. Thus, the Fgf10(iCre) line should serve as a powerful tool to explore the functions of Fgf10 in a controlled and stage-specific manner.
German Center for Lung Research, Excellence Cluster Cardio-Pulmonary System, Universities of Giessen and Marburg Lung Center, Giessen, Hessen, Germany. saverio.bellusci@innere.med.uni-giessen.de
Elie
E
El Agha
Denise
D
Al Alam
Gianni
G
Carraro
Breanne
B
MacKenzie
Kerstin
K
Goth
Stijn P
SP
De Langhe
Robert
R
Voswinckel
Mohammad K
MK
Hajihosseini
Virender K
VK
Rehan
Saverio
S
Bellusci
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D047108:Embryonic Development; D005091:Exons; D051526:Fibroblast Growth Factor 10 / Q000235:genetics*; D020868:Gene Silencing; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D016133:Polymerase Chain Reaction; D013629:Tamoxifen / Q000008:administration & dosage
D017931:DNA Primers; C496464:Fgf10 protein, mouse; D051526:Fibroblast Growth Factor 10; D013629:Tamoxifen