AUTHOR=Jones Matthew R. , Lingampally Arun , Dilai Salma , Shrestha Amit , Stripp Barry , Helmbacher Francoise , Chen Chengshui , Chao Cho-Ming , Bellusci Saverio 

TITLE=Characterization of Tg(Etv4-GFP) and Etv5RFP Reporter Lines in the Context of Fibroblast Growth Factor 10 Signaling During Mouse Embryonic Lung Development

JOURNAL=Frontiers in Genetics

VOLUME=10

YEAR=2019

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2019.00178

DOI=10.3389/fgene.2019.00178

ISSN=1664-8021

ABSTRACT=<p>Members of the PEA3 transcription factors are emerging as bone fide targets for fibroblast growth factor (FGF) signaling. Among them, ETV4 and ETV5 appear to mediate FGF10 signaling during early embryonic lung development. In this paper, recently obtained <italic>Tg(Etv4-GFP)</italic> and <italic>Etv5</italic><sup><italic>CreERT</italic>2−<italic>RFP</italic></sup> fluorescent reporter lines were generally characterized during early embryonic development and in the context of FGF10 signaling, in particular. We found that both <italic>Tg(Etv4-GFP)</italic> and <italic>Etv5</italic><sup><italic>CreERT</italic>2−<italic>RFP</italic></sup> were primarily expressed in the epithelium of the lung during embryonic development. However, the expression of <italic>Etv5</italic><sup><italic>CreERT</italic>2−<italic>RFP</italic></sup> was much higher than that of <italic>Tg(Etv4-GFP)</italic>, and continued to increase during development, whereas <italic>Tg(Etv4-GFP)</italic> decreased. The expression patterns of the surrogate fluorescent protein GFP and RFP for ETV4 and ETV5, respectively, agreed with known regions of FGF10 signaling in various developing organs, including the lung, where ETV4-GFP was seen primarily in the distal epithelium and to a lesser extent in the surrounding mesenchyme. As expected, ETV5-RFP was restricted to the lung epithelium, showing a decreasing expression pattern from distal buds to proximal conducting airways. FGF10 inhibition experiments confirmed that both <italic>Etv4</italic> and <italic>Etv5</italic> are downstream of FGF10 signaling. Finally, we also validated that both fluorescent reporters responded to FGF10 inhibition <italic>in vitro</italic>. In conclusion, these two reporter lines appear to be promising tools to monitor FGF10/FGFR2b signaling in early lung development. These tools will have to be further validated at later stages and in other organs of interest.</p>