Cadherin-6B is proteolytically processed during epithelial-to-mesenchymal transitions of the cranial neural crest

Schiffmacher, Andrew T.; Padmanabhan, Rangarajan; Jhingory, Sharon; Taneyhill, Lisa A.

doi:10.1091/mbc.e13-08-0459

Cited by 60 publications

(137 citation statements)

References 66 publications

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“…Since junctions were enhanced when ADAMTS6 was depleted or mutated, ADAMTS6 may degrade them. Indeed, other ADAMs can cleave cadherins37. Disruption of tight junctions was not seen with overexpression of the two ADAMTS6 mutants, confirming that the metalloprotease activity is responsible for tight junction disruption.…”

Section: Discussionmentioning

confidence: 81%

ADAMTS-10 and -6 differentially regulate cell-cell junctions and focal adhesions

Cain

Mularczyk

Singh

et al. 2016

Sci Rep

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ADAMTS10 and ADAMTS6 are homologous metalloproteinases with ill-defined roles. ADAMTS10 mutations cause Weill-Marchesani syndrome (WMS), implicating it in fibrillin microfibril biology since some fibrillin-1 mutations also cause WMS. However little is known about ADAMTS6 function. ADAMTS10 is resistant to furin cleavage, however we show that ADAMTS6 is effectively processed and active. Using siRNA, over-expression and mutagenesis, it was found ADAMTS6 inhibits and ADAMTS10 is required for focal adhesions, epithelial cell-cell junction formation, and microfibril deposition. Either knockdown of ADAMTS6, or disruption of its furin processing or catalytic sites restores focal adhesions, implicating its enzyme activity acts on targets in the focal adhesion complex. In ADAMTS10-depleted cultures, expression of syndecan-4 rescues focal adhesions and cell-cell junctions. Recombinant C-termini of ADAMTS10 and ADAMTS6, both of which induce focal adhesions, bind heparin and syndecan-4. However, cells overexpressing full-length ADAMTS6 lack heparan sulphate and focal adhesions, whilst depletion of ADAMTS6 induces a prominent glycocalyx. Thus ADAMTS10 and ADAMTS6 oppositely affect heparan sulphate-rich interfaces including focal adhesions. We previously showed that microfibril deposition requires fibronectin-induced focal adhesions, and cell-cell junctions in epithelial cultures. Here we reveal that ADAMTS6 causes a reduction in heparan sulphate-rich interfaces, and its expression is regulated by ADAMTS10.

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Section: Discussionmentioning

confidence: 81%

ADAMTS-10 and -6 differentially regulate cell-cell junctions and focal adhesions

Cain

Mularczyk

Singh

et al. 2016

Sci Rep

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“…Protein extraction and immunoblotting was performed as described in (Schiffmacher et al, 2014). Neural tubes electroporated with either the Annexin A6 or control MO were excised 6–8 hours post-electroporation, pooled, pelleted, flash-frozen in liquid nitrogen, and stored at −80 C until needed for immunoblot analysis.…”

Section: Methodsmentioning

confidence: 99%

“…Measurements of each of the two morphological types of neurons (control MO: bipolar; Annexin A6 MO: short projections) were performed using the Zeiss Zen 2.0 software from Z-stacks generated from at least five serial sections through a minimum of five each control MO- and Annexin A6 MO-electroporated embryos. Results are reported as means +/− standard deviation and standard error of the mean, with statistical significance of data established using an unpaired student’s t test (Schiffmacher et al, 2014) (Wu et al, 2014). …”

Section: Methodsmentioning

confidence: 99%

Annexin A6 controls neuronal membrane dynamics throughout chick cranial sensory gangliogenesis

Shah

Schiffmacher

Taneyhill

2017

Developmental Biology

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Cranial sensory ganglia are components of the peripheral nervous system that possess a significant somatosensory role and include neurons within the trigeminal and epibranchial nerve bundles. Although it is well established that these ganglia arise from interactions between neural crest and neurogenic placode cells, the molecular basis of ganglia assembly is still poorly understood. Members of the Annexin protein superfamily play key roles in sensory nervous system development throughout metazoans. Annexin A6 is expressed in chick trigeminal and epibranchial placode cell-derived neuroblasts and neurons, but its function in cranial ganglia formation has not been elucidated. To this end, we interrogated the role of Annexin A6 using gene perturbation studies in the chick embryo. Our data reveal that placode cell-derived neuroblasts with reduced Annexin A6 levels ingress and migrate normally to the ganglionic anlage, where neural crest cell corridors correctly form around them. Strikingly, while Annexin A6-depleted placode cell-derived neurons still express mature neuronal markers, they fail to form two long processes, which are considered morphological features of mature neurons, and no longer innervate their designated targets due to the absence of this bipolar morphology. Moreover, overexpression of Annexin A6 causes some placode cell-derived neurons to form extra protrusions alongside these bipolar processes. These data demonstrate that the molecular program associated with neuronal maturation is distinct from that orchestrating changes in neuronal morphology, and, importantly, reveal Annexin A6 to be a key membrane scaffolding protein during sensory neuron membrane biogenesis. Collectively, our results provide novel insight into mechanisms underscoring morphological changes within placode cell-derived neurons that are essential for cranial gangliogenesis.

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“…Ncad upregulation is thought to promote EMT in several nonneuroepithelial cells (reviewed by Wheelock et al, 2008), but in many cases Ncad expression in epithelial cells does not correlate with EMT (Dady et al, 2012;Knudsen et al, 2005;Maeda et al, 2005;Straub et al, 2011). Interestingly, it has been shown that Cdh6 also undergoes proteolytic cleavage during EMT (Schiffmacher et al, 2014), which may be one of the post-translational mechanisms by which Cdh6 is regulated during EMT. It will be interesting to see whether Cdh6 cleavage fragments also carry signaling abilities similar to Ncad or to other type II cadherins (Cad7 and Cad11), the signaling functions of which promote NCC migration (Kashef et al, 2009;McCusker et al, 2009;Nakagawa and Takeichi, 1998;Vallin et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

Cadherin-6 promotes neural crest cell detachment via F-actin regulation and influences active Rho distribution during epithelial to mesenchymal transition

Clay¹,

Halloran²

2014

Journal of Cell Science

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The epithelial-to-mesenchymal transition (EMT) is a complex change in cell phenotype that is important for cell migration, morphogenesis and carcinoma metastasis. Loss of epithelial cell adhesion and tight regulation of cadherin adhesion proteins are crucial for EMT. Cells undergoing EMT often display cadherin switching, where they downregulate one cadherin and induce expression of another. However, the functions of the upregulated cadherins and their effects on cell motility are poorly understood. Neural crest cells (NCCs), which undergo EMT during development, lose N-cadherin and upregulate Cadherin 6 (Cdh6) prior to EMT. Cdh6 has been suggested to suppress EMT via cell adhesion, but also to promote EMT by mediating pro-EMT signals. Here, we determine novel roles for Cdh6 in generating cell motility during EMT. We use live imaging of NCC behavior in vivo to show that Cdh6 promotes detachment of apical NCC tails, an important early step of EMT. Furthermore, we show that Cdh6 affects spatiotemporal dynamics of F-actin and active Rho GTPase, and that Cdh6 is required for accumulation of F-actin in apical NCC tails during detachment. Moreover, Cdh6 knockdown alters the subcellular distribution of active Rho, which is known to promote localized actomyosin contraction that is crucial for apical NCC detachment. Together, these data suggest that Cdh6 is an important determinant of where subcellular actomyosin forces are generated during EMT. Our results also identify mechanisms by which an upregulated cadherin can generate cell motility during EMT.

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Cadherin-6B is proteolytically processed during epithelial-to-mesenchymal transitions of the cranial neural crest

Cited by 60 publications

References 66 publications

ADAMTS-10 and -6 differentially regulate cell-cell junctions and focal adhesions

ADAMTS-10 and -6 differentially regulate cell-cell junctions and focal adhesions

Annexin A6 controls neuronal membrane dynamics throughout chick cranial sensory gangliogenesis

Cadherin-6 promotes neural crest cell detachment via F-actin regulation and influences active Rho distribution during epithelial to mesenchymal transition

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