Leukemia inhibitory factor in rat fetal lung development: expression and functional studies

Conceived and designed the experiments: CN-S EC-D JC-P. Performed the experiments: CN-S PP EC-D FOP RSM. Analyzed the data: CN-S PP JC-P. Wrote the paper: CN-S PP RSM JC-P.Leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) are members of the family of the glycoprotein 130 (gp130)-type cytokines. These cytokines share gp130 as a common signal transducer, which explains why they show some functional redundancy. Recently, it was demonstrated that IL-6 promotes fetal lung branching. Additionally, LIF has been implicated in developmental processes of some branching organs. Thus, in this study LIF expression pattern and its effects on fetal rat lung morphogenesis were assessed. Methodology/Principal Findings LIF and its subunit receptor LIFRα expression levels were evaluated by immunohistochemistry and western blot in fetal rat lungs of different gestational ages, ranging from 13.5 to 21.5 days post-conception. Throughout all gestational ages studied, LIF was constitutively expressed in pulmonary epithelium, whereas LIFRα was first mainly expressed in the mesenchyme, but after pseudoglandular stage it was also observed in epithelial cells. These results point to a LIF epithelium-mesenchyme cross-talk, which is known to be important for lung branching process. Regarding functional studies, fetal lung explants were cultured with increasing doses of LIF or LIF neutralizing antibodies during 4 days. MAPK, AKT, and STAT3 phosphorylation in the treated lung explants was analyzed. LIF supplementation significantly inhibited lung growth in spite of an increase in p44/42 phosphorylation. On the other hand, LIF inhibition significantly stimulated lung growth via p38 and Akt pathways. Conclusions/Significance The present study describes that LIF and its subunit receptor LIFRα are constitutively expressed during fetal lung development and that they have an inhibitory physiological role on fetal lung branching.This project was funded by Fundacao para a Ciencia e a Tecnologia (PTDC/SAU-OBD/108051/2008) and by Seccao de Neonatologia da Sociedade Portuguesa de Pediatria (Grant ZERU 2008). RSM was supported by Fundacao para a Ciencia e a Tecnologia (reference SFRH/BPD/15408/2005). FOP was supported by Fundacao para a Ciencia e a Tecnologia (reference UMINHO/BII/172/2009). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript