Maintenance of the Epithelial Barrier and Remodeling of Cell-Cell Junctions during Cytokinesis

Abstract

Summary Epithelial integrity and barrier function must be maintained during the complex cell shape changes that occur during cytokinesis in vertebrate epithelial tissue. Here, we investigate how adherens junctions and bicellular and tricellular tight junctions are maintained and remodeled during cell division in the Xenopus laevis embryo. We find that epithelial barrier function is not disrupted during cytokinesis and is mediated by sustained tight junctions. Using FRAP, we demonstrate that adherens junction proteins are stabilized at the cleavage furrow by increased tension. We find that vinculin is recruited to the adherens junction at the cleavage furrow, and inhibiting recruitment of vinculin by expressing a dominant negative mutant increases the rate of furrow ingression. Furthermore, we show that cells neighboring the cleavage plane are pulled between the daughter cells, making a new interface between neighbors, and two new tricellular tight junctions flank the midbody following cytokinesis. Our data provide new insight into how epithelial integrity and barrier function are maintained throughout cytokinesis in vertebrate epithelial tissue.

RhoA regulates actin network dynamics during apical surface emergence in multiciliated epithelial cells

“…Epithelial cell turnover poses unique challenges, and simply maintaining a fluid-tight barrier during cell division requires a tight choreography between cells (for example, see Higashi et al, 2016). More complex epithelia face even greater difficulties.…”

Rho GTPases and actomyosin: Partners in regulating epithelial cell-cell junction structure and function

“…FRAP studies have proven to be a powerful tool to tease apart junction protein complexes, their interactions with the cytoskeleton, and how junctions remodel in response to stimuli such as changes in tension (Priya et al, 2013; Ratheesh et al, 2012; Van Itallie et al, 2016; Yonemura et al, 2010). For example, FRAP was recently used to demonstrated that plasticity of cell-cell junction structure and the linkage to actin is important for maintenance of cell-cell adhesion and barrier function when cells undergo cell-cell junction remodeling during cell division in Xenopus embryos (Higashi et al, 2016). This study showed that tension generated by the cytokinetic contractile ring is transmitted to the adherens junction, recruiting Vinculin, and stabilizing the dynamics of adherens junction proteins specifically at the division site (Higashi et al, 2016).…”

“…For example, FRAP was recently used to demonstrated that plasticity of cell-cell junction structure and the linkage to actin is important for maintenance of cell-cell adhesion and barrier function when cells undergo cell-cell junction remodeling during cell division in Xenopus embryos (Higashi et al, 2016). This study showed that tension generated by the cytokinetic contractile ring is transmitted to the adherens junction, recruiting Vinculin, and stabilizing the dynamics of adherens junction proteins specifically at the division site (Higashi et al, 2016). …”

“…Indeed, a recent study in Xenopus epithelial cells, demonstrates implementation of both of these mechanisms. When increased tension was generated by a physiological mechanical input – tension generated by the contractile ring during cell division – embryos exhibited a stabilization of AJ proteins and recruited Vinculin to the division site to strengthen the AJ in response to locally applied tension (Higashi et al, 2016). …”

Living Xenopus oocytes, eggs, and embryos as models for cell division

“…Similarly, local activation of Rho GTPases and cytoskeletal mobilization during cell repair were first described in Xenopus oocytes and embryos (Benink & Bement, 2005; Clark et al, 2009; Mandato & Bement, 2001, 2003) and subsequently demonstrated for repair of many different cell types (Abreu-Blanco, Verboon, & Parkhurst, 2011, 2014; Kono, Saeki, Yoshida, Tanaka, & Pellman, 2012; Lin et al, 2012; McDade, Archambeau, & Michele, 2014; Nakamura, Verboon, & Parkhurst, 2017). More recently, live imaging of early Xenopus embryos has been used to investigate the dynamics of epithelial cell–cell junctions (Higashi, Arnold, Stephenson, Dinshaw, & Miller, 2016; Reyes et al, 2014), formation of cilia (Chung et al, 2014; Kim et al, 2010), and dynamic remodeling of the epithelial layer (Sedzinski, Hannezo, Tu, Biro, & Wallingford, 2016). In short, if a given cellular process occurs within several micrometers of the embryo surface, this model can be used to study it.…”