A fetal mouse lung slice model for molecular live cell imaging of the pulmonary neuroepithelial body microenvironment

Abstract

The prominent presence of differentiated neuroepithelial bodies (NEBs) from early embryonic development on, with the highest density in the perinatal period, reflects important prenatal functions and involvement in the transition from fetal to neonatal life. We therefore adapted our established ex vivo live cell imaging lung slice model to allow functional studies in fetal mouse lungs.Using Fluo‐4, changes in [Ca2+]i could be monitored, revealing that ATP causes a [Ca2+]i rise in fetal airway epithelial cells. Additional identification of prenatal NEBs with the vital dye 4‐Di‐2‐ASP, showed that extracellular high K+ evoked a reversible [Ca2+]i increase in fetal NEB cells. Fixed lung slices of prenatal mice revealed immunostaining for CGRP in NEBs, and a P2X3 ATP receptor‐expressing population of NEB‐associated nerve terminals, highlighting the possibility to directly link physiology to neurochemistry in this model. Since slices of fetal GAD67‐GFP knock‐in mice show selective GFP fluorescence in NEB cells, use of these animals will definitely improve physiological studies of NEBs.The presented fetal mouse lung slice models offer excellent possibilities for molecular live cell imaging to further unravel the functional significance of the NEB microenvironment during the pre‐ and perinatal period.Grants: IWT (SB 81162); FWO (G.0589.11), Hercules Foundation (AUAH‐09–001); UA (GOA BOF 2007; KP BOF 2011)