Spatial and temporal PCP protein dynamics coordinate cell intercalation during neural tube closure

AbstractPlanar cell polarity (PCP) controls the convergent extension cell movements that drive axis elongation in all vertebrates. Though asymmetric localization of core PCP proteins is central to their function, we currently understand little about PCP protein localization as it relates to the subcellular behaviors that drive convergent extension. Here, we have used high magnification time-lapse imaging to simultaneously monitor cell intercalation behaviors and the localization of the PCP proteins Prickle2 and Vangl2. We observed the expected asymmetric enrichment of PCP proteins, but more interestingly, we also observed tight temporal and spatial correlation of PCP protein enrichment with contractile behavior in cell-cell junctions. These patterns of localization were associated with similar pattern of protein turnover at junctions as assessed by FRAP. In fact, dynamic enrichment of PCP proteins was linked more strongly to junction behavior than to spatial orientation. Finally, recruitment of Prickle2 and Vangl2 to cell-cell junctions was temporally and spatially coordinated with planar polarized oscillations of actomyosin enrichment, and all of these dynamic relationships were disrupted when PCP signaling was manipulated. Together, these results provide a dynamic and quantitative view of PCP protein localization during convergent extension and suggest a complex and intimate link between the dynamic localization of core PCP proteins, actomyosin assembly, and polarized junction shrinking during cell intercalation of the closing vertebrate neural tube.

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Spatial and temporal analysis of PCP protein dynamics during neural tube closure

eLife ◽

10.7554/elife.36456 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 13

Author(s):

Mitchell T Butler ◽

John B Wallingford

Keyword(s):

Protein Dynamics ◽

Neural Tube ◽

Planar Cell Polarity ◽

Protein Localization ◽

Temporal Analysis ◽

Neural Tube Closure ◽

Cell Junctions ◽

Convergent Extension ◽

Turnover Rates ◽

Cell Intercalation

Planar cell polarity (PCP) controls convergent extension and axis elongation in all vertebrates. Although asymmetric localization of PCP proteins is central to their function, we understand little about PCP protein localization during convergent extension. Here, we use quantitative live imaging to simultaneously monitor cell intercalation behaviors and PCP protein dynamics in the Xenopus laevis neural plate epithelium. We observed asymmetric enrichment of PCP proteins, but more interestingly, we observed tight correlation of PCP protein enrichment with actomyosin-driven contractile behavior of cell-cell junctions. Moreover, we found that the turnover rates of junctional PCP proteins also correlated with the contractile behavior of individual junctions. All these dynamic relationships were disrupted when PCP signaling was manipulated. Together, these results provide a dynamic and quantitative view of PCP protein localization during convergent extension and suggest a complex and intimate link between the dynamic localization of core PCP proteins, actomyosin assembly, and polarized junction shrinking during cell intercalation in the closing vertebrate neural tube.

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Mechanical heterogeneity along single cell-cell junctions is driven by lateral clustering of cadherins during vertebrate axis elongation

eLife ◽

10.7554/elife.65390 ◽

2021 ◽

Vol 10 ◽

Author(s):

Robert J Huebner ◽

Abdul Naseer Malmi-Kakkada ◽

Sena Sarikaya ◽

Shinuo Weng ◽

D Thirumalai ◽

...

Keyword(s):

Cell Biology ◽

Planar Cell Polarity ◽

Protein Localization ◽

Tissue Level ◽

Cell Junctions ◽

Convergent Extension ◽

Length Scales ◽

Mechanical Heterogeneity ◽

Multiple Length Scales ◽

Cell Cell

Morphogenesis is governed by the interplay of molecular signals and mechanical forces across multiple length scales. The last decade has seen tremendous advances in our understanding of the dynamics of protein localization and turnover at sub-cellular length scales, and at the other end of the spectrum, of mechanics at tissue-level length scales. Integrating the two remains a challenge, however, because we lack a detailed understanding of the subcellular patterns of mechanical properties of cells within tissues. Here, in the context of the elongating body axis of Xenopus embryos, we combine tools from cell biology and physics to demonstrate that individual cell-cell junctions display finely-patterned local mechanical heterogeneity along their length. We show that such local mechanical patterning is essential for the cell movements of convergent extension and is imparted by locally patterned clustering of a classical cadherin. Finally, the patterning of cadherins and thus local mechanics along cell-cell junctions are controlled by Planar Cell Polarity signaling, a key genetic module for CE that is mutated in diverse human birth defects.

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Mechanical heterogeneity along single cell-cell junctions is driven by lateral clustering of cadherins during vertebrate axis elongation

10.1101/2020.02.11.944033 ◽

2020 ◽

Cited By ~ 1

Author(s):

Robert J. Huebner ◽

Abdul Naseer Malmi-Kakkada ◽

Sena Sarikaya ◽

Shinuo Weng ◽

D. Thirumalai ◽

...

Keyword(s):

Cell Biology ◽

Planar Cell Polarity ◽

Protein Localization ◽

Tissue Level ◽

Cell Junctions ◽

Convergent Extension ◽

Length Scales ◽

Mechanical Heterogeneity ◽

Multiple Length Scales ◽

Cell Cell

AbstractMorphogenesis is governed by the interplay of molecular signals and mechanical forces across multiple length scales. The last decade has seen tremendous advances in our understanding of the dynamics of protein localization and turnover at sub-cellular length scales, and at the other end of the spectrum, of mechanics at tissue-level length scales. Integrating the two remains a challenge, however, because we lack a detailed understanding of the subcellular patterns of mechanical properties of cells within tissues. Here, in the context of the elongating body axis of a vertebrate embryo, we combine tools from cell biology and physics to demonstrate that individual cell-cell junctions display finely-patterned local mechanical heterogeneity along their length. We show that such local mechanical patterning is essential for the cell movements of convergent extension and is imparted by locally patterned clustering of a classical cadherin. Finally, the patterning of cadherins and thus local mechanics along cell-cell junctions are controlled by Planar Cell Polarity signaling, a key genetic module for CE that is mutated in diverse human birth defects.

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Severe neural tube defects due to failure of closure initiation can arise without abnormality of neuroepithelial convergent extension

10.1101/2021.07.04.451044 ◽

2021 ◽

Author(s):

Oleksandr Nychyk ◽

Gabriel L Galea ◽

Matteo J Mole ◽

Dawn Savery ◽

Nicholas Greene ◽

...

Keyword(s):

Neural Tube ◽

Planar Cell Polarity ◽

Gene Mutations ◽

Initiation Site ◽

Positive Control ◽

Severe Form ◽

Neural Tube Closure ◽

Null Alleles ◽

Convergent Extension ◽

Partial Loss

Planar cell polarity (PCP) signalling is vital for initiation of neural tube closure in mice, with diminished convergent extension (CE) cell movements leading to a severe form of neural tube defect (NTD), termed craniorachischisis (CRN). Some human NTDs are also associated with PCP gene mutations, but affected individuals are generally heterozygous, whereas PCP homozygosity or compound heterozygosity is needed to produce CRN in mice. This suggests human NTDs may involve other genetic or environmental factors, that interact with partial loss of PCP function. We found that reduced sulfation OF glycosaminoglycans (GAGs) interacts with heterozygosity for the Lp allele of Vangl2 (a core PCP gene), to cause CRN in mice. Here, we hypothesised that this GAG-PCP interaction may regulate convergent extension movements, and hence lead to severe NTDs in the context of only partial loss of PCP function. Both Lp and null alleles of Vangl2 gave similar findings. Culture of E8.5 embryos in the presence of chlorate (a GAG sulfation inhibitor), or enzymatic cleavage of GAG chains, led to failure of NT closure initiation in the majority of Lp/+ embryos, whereas few +/+ littermates exhibited CRN. The chlorate effect was rescued by exogenous sulphate. Surprisingly, DiO labeling of the embryonic node demonstrated no abnormality of midline axial extension in chlorate-treated Lp/+ embryos that developed CRN. In contrast, positive control Lp/Lp embryos displayed severe convergent extension defects in this assay. Morphometric analysis of the closure initiation site revealed abnormalities in the size and shape of somites that flank the closing neural tube in chlorate-treated Lp/+ embryos. We conclude that severe NTDs involving failure of closure initiation can arise by a mechanism other than faulty neuroepithelial convergent extension. Matrix-mediated expansion of somites, flanking the closing neural tube, may be required for closure initiation.

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Convergent extension, planar-cell-polarity signalling and initiation of mouse neural tube closure

Development ◽

10.1242/dev.000380 ◽

2007 ◽

Vol 134 (4) ◽

pp. 789-799 ◽

Cited By ~ 185

Author(s):

P. Ybot-Gonzalez ◽

D. Savery ◽

D. Gerrelli ◽

M. Signore ◽

C. E. Mitchell ◽

...

Keyword(s):

Cell Polarity ◽

Neural Tube ◽

Planar Cell Polarity ◽

Neural Tube Closure ◽

Convergent Extension ◽

Tube Closure ◽

Planar Cell Polarity Signalling

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Update on the Role of the Non-Canonical Wnt/Planar Cell Polarity Pathway in Neural Tube Defects

Cells ◽

10.3390/cells8101198 ◽

2019 ◽

Vol 8 (10) ◽

pp. 1198 ◽

Cited By ~ 5

Author(s):

Wang ◽

Marco ◽

Capra ◽

Kibar

Keyword(s):

Cell Polarity ◽

Neural Tube ◽

Neural Tube Defects ◽

Planar Cell Polarity ◽

Neural Tube Closure ◽

Convergent Extension ◽

Complex Genetics ◽

Pcp Signaling ◽

Canonical Wnt

Neural tube defects (NTDs), including spina bifida and anencephaly, represent the most severe and common malformations of the central nervous system affecting 0.7–3 per 1000 live births. They result from the failure of neural tube closure during the first few weeks of pregnancy. They have a complex etiology that implicate a large number of genetic and environmental factors that remain largely undetermined. Extensive studies in vertebrate models have strongly implicated the non-canonical Wnt/planar cell polarity (PCP) signaling pathway in the pathogenesis of NTDs. The defects in this pathway lead to a defective convergent extension that is a major morphogenetic process essential for neural tube elongation and subsequent closure. A large number of genetic studies in human NTDs have demonstrated an important role of PCP signaling in their etiology. However, the relative contribution of this pathway to this complex etiology awaits a better picture of the complete genetic architecture of these defects. The emergence of new genome technologies and bioinformatics pipelines, complemented with the powerful tool of animal models for variant interpretation as well as significant collaborative efforts, will help to dissect the complex genetics of NTDs. The ultimate goal is to develop better preventive and counseling strategies for families affected by these devastating conditions.

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Global analysis of cell behavior and protein localization dynamics reveals region-specific functions for Shroom3 and N-cadherin during neural tube closure

10.1101/2021.01.19.427312 ◽

2021 ◽

Author(s):

Austin T. Baldwin ◽

Juliana Kim ◽

John B. Wallingford

Keyword(s):

Neural Tube ◽

Cell Biology ◽

Protein Localization ◽

Global Analysis ◽

Time Lapse ◽

Tissue Level ◽

Neural Tube Closure ◽

Cell Behaviors ◽

Regional Specificity ◽

Tube Closure

AbstractFailures of neural tube closure are common and serious birth defects, yet we have a poor understanding of the interaction of genetics and cell biology during neural tube closure. Additionally, mutations that cause neural tube defects (NTDs) tend to affect anterior or posterior regions of the neural tube but rarely both, indicating a regional specificity to NTD genetics. To better understand the regional specificity of cell behaviors during neural tube closure, we analyzed the dynamic localization of actin and N-cadherin via high-resolution tissue-level time-lapse microscopy during Xenopus neural tube closure. To investigate the regionality of gene function, we generated mosaic mutations in shroom3, a key regulator or neural tube closure This approach elucidates new differences between cell behaviors during cranial/anterior and spinal/posterior neural tube closure, provides mechanistic insight into the function of shroom3 and demonstrates the ability of tissue-level imaging and analysis to generate cell-biological mechanistic insights into neural tube closure.

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