The Effects of Maternal Stress on TLR Expression in Neonatal Mus musculus Alveolar Epithelial Cells

Abstract

Prior research has confirmed an association between maternal stress and offspring asthma. Recent studies suggest a role for epithelial expressed toll-like receptors (TLRs) in allergic asthma development. In this study, we were interested in isolating alveolar epithelial cells (AECs) from neonatal mice and analyzing how maternal stress potentially changes the expression of TLRs in the AECs. C57bl/6J strain mice were mated and pregnant dams started treatment at midgestation (E12.5) with free access to Vehicle or CORT-treated drinking water until the litter was born. The litter was sacrificed on postnatal day 7 (7 PND). The lungs were harvested and both AECs (EpCAM+) and immune cells (CD45+) were isolated based on staining with epithelial cell adhesion molecule (EpCAM) , CD45, and CD31 fluorescent antibodies. Here we present our optimized protocol for isolating EpCAM+ and CD45+ cells in 7 PND neonates through fluorescence activated cell sorting (FACS). Further experiments to investigate TLR expression in 7 PND AECs will be conducted. Also, we plan to further explore alveolar immune cell populations in the neonates.