Junctional Adhesion Molecule 3 Expression in the Mouse Airway Epithelium Is Linked to Multiciliated Cells

Clara Maria Mateos-Quiros,Jose Maria Carvajal-Gonzalez,Javier Francisco-Morcillo,Juan Francisco Barrera-Lopez,Selene Diaz-Chamorro,Guadalupe Álvarez-Hernán,Sergio Garrido-Jimenez,Francisco Centeno,Angel Carlos Roman

doi:10.3389/fcell.2021.622515

Clara Maria Mateos-Quiros, Jose Maria Carvajal-Gonzalez + Show 7 more

Open Access

https://doi.org/10.3389/fcell.2021.622515

Copy DOI

Abstract

Tight-junction (TJ) proteins are essential for establishing the barrier function between neighbor epithelial cells, but also for recognition of pathogens or cell migration. Establishing the expression pattern and localization of different TJ proteins will help to understand the development and physiology of the airway. Here we identify that the junctional adhesion molecule 3 (Jam3) expression is restricted to multiciliated cells (MCCs) in the airway epithelium. In vitro, Jam3 expression varies along airway basal stem cell (BSC) differentiation and upon DAPT treatment or IL6 exposure. However, Jam3 is not required for BSC differentiation to specific cell types. In addition, we found that MCC lacking Jam3 display normal cilia morphology and cilia beating frequency with a delay in BB assembly/positioning in MCCs during differentiation. Remarkably, Jam3 in MCC is mostly localized to subapical organelles, which are negative for the apical recycling endosome marker Rab11 and positive for EEA1. Our data show that Jam3 expression is connected to mature MCC in the airway epithelium and suggest a Jam3 role unrelated to its known barrier function.

Highlights

Junctional adhesion molecule 3 (Jam3), together with Jam1 and Jam2, is known as a tight-junction (TJ) component that belongs to the junctional adhesion molecule family of proteins (Hartmann et al, 2020)
A costaining with Jam3 and acetylated tubulin, which accumulates in the cilium axoneme, in a whole-mount trachea showed that Jam3 was co-localizing with multiciliated cells (MCCs) labeled with acetylated tubulin (Figures 1B–B”’)
We noticed that in our whole-mount confocal images, Jam3 expression levels were heterogeneous among MCCs, where we can find a mix of cells with low or high Jam3 levels (Figures 1C–C”)

Summary

Introduction

Junctional adhesion molecule 3 (Jam3), together with Jam and Jam, is known as a tight-junction (TJ) component that belongs to the junctional adhesion molecule family of proteins (Hartmann et al, 2020). Since its discovery in 2004 (Cunningham et al, 2000), various in vitro studies have suggested that Jam has important functions in the assembly of TJs in endothelial and epithelial cells, and in transendothelial and transepithelial migration of immune cells (Bazzoni, 2003; Ebnet, 2017; Hartmann et al, 2020). Jam has been proposed to promote neutrophil migration in vivo and in vitro in endothelial cells (Chavakis et al, 2004; Woodfin et al, 2011). This Jam3-dependent neutrophil migration phenotype was described in vitro with epithelial cells, where Jam is a component of desmosomes (Zen et al, 2004). Interleukin 13 (IL13), which drives asthma symptoms or IL-4 and IL-13 enhanced by IFN-gamma, induces changes in the TJ protein composition so that paracellular permeability becomes increased (Ahdieh et al, 2001; Schmidt et al, 2019)

Methods

Results

Conclusion