ADAM13 cleavage of cadherin-11 promotes CNC migration independently of the homophilic binding site

Abstract

The cranial neural crest (CNC) is a highly motile population of cells that is responsible for forming the face and jaw in all vertebrates and perturbing their migration can lead to craniofacial birth defects. Cell motility requires a dynamic modification of cell–cell and cell-matrix adhesion. In the CNC, cleavage of the cell adhesion molecule cadherin-11 by ADAM13 is essential for cell migration. This cleavage generates a shed extracellular fragment of cadherin-11 (EC1-3) that possesses pro-migratory activity via an unknown mechanism. Cadherin-11 plays an important role in modulating contact inhibition of locomotion (CIL) in the CNC to regulate directional cell migration. Here, we show that while the integral cadherin-11 requires the homophilic binding site to promote CNC migration in vivo, the EC1-3 fragment does not. In addition, we show that increased ADAM13 activity or expression of the EC1-3 fragment increases CNC invasiveness in vitro and blocks the repulsive CIL response in colliding cells. This activity requires the presence of an intact homophilic binding site on the EC1-3 suggesting that the cleavage fragment may function as a competitive inhibitor of cadherin-11 adhesion in CIL but not to promote cell migration in vivo.

Cadherins function during the collective cell migration of Xenopus Cranial Neural Crest cells: revisiting the role of E-cadherin

“…The cadherin-11 extracellular domain is cleaved by the metalloproteases ADAM9 and 13 generating a fragment EC 1–3 containing the first 3 cadherin repeats, that is necessary for CNC migration (McCusker et al, 2009). This fragment can only be detected at the onset of migration and the replacement of endogenous cadherin-11 by an engineered uncleavable form of the protein does not support CNC migration, demonstrating that cadherin-11 cleavage is essential for neural crest migration (Abbruzzese et al, 2016; McCusker et al, 2009). Since the homophilic binding domain of EC1–3 is dispensable for CNC migration, it is unlikely that EC1–3 acts as an antagonist of homophilic interactions between CNC cells (Abbruzzese et al, 2016).…”

“…This fragment can only be detected at the onset of migration and the replacement of endogenous cadherin-11 by an engineered uncleavable form of the protein does not support CNC migration, demonstrating that cadherin-11 cleavage is essential for neural crest migration (Abbruzzese et al, 2016; McCusker et al, 2009). Since the homophilic binding domain of EC1–3 is dispensable for CNC migration, it is unlikely that EC1–3 acts as an antagonist of homophilic interactions between CNC cells (Abbruzzese et al, 2016). It is interesting to note that ADAM metalloproteases can also shed N- and E-cadherin extracellular domains in vitro (Najy et al, 2008; Reiss et al, 2005) and that these N-terminal fragments also have biological properties such as activating Receptor Tyrosine Kinases (Cifuentes-Diaz et al, 1994; Najy et al, 2008; Paradies and Grunwald, 1993).…”

“…It is therefore possible that the cadherin-11 EC1–3 fragment may do the same. In addition, the depletion of the extracellular domain of cadherin-11 or the overexpression of the EC1–3 fragment in Xenopus leads to loss of CIL and directionality of the CNC (Abbruzzese et al, 2016; Becker et al, 2013). The intracellular domain of cadherin-11 has also a crucial function for CNC migration.…”

Animal models of craniofacial anomalies

“…Xenopus is a system that is very amenable to molecular and physical manipulation (e.g. microdissection), as well as cellular and biochemical assays (Abbruzzese et al, 2015;Devotta et al, 2016). These are also historically easily coupled with large scale screening (Grammer et al, 2000).…”

Resolving in vivo gene expression during collective cell migration using an integrated RNAscope, immunohistochemistry and tissue clearing method

“…In vivo time-lapse imaging has revealed the dynamics of group cell behaviors in a number of embryo model organisms and included early gonad development (Irizarry and Stathopoulos, 2015; Paksa et al, 2016), gastrulation (Campinho et al, 2013; Omelchenko et al, 2014) hindgut formation (Nerurkar et al, 2017) and craniofacial patterning (Abbruzzese et al, 2016; Powell et al, 2015). However, our understanding of the molecular choreography underlying collective cell migration events has remained challenging due to difficulties with in vivo interrogation of gene and protein expression in neighboring cells and deep within the intact 3D embryo.…”