Defining domain-specific orientational order in the desmosomal cadherins

Dean, William F.; Mattheyses, Alexa L.

doi:10.1016/j.bpj.2022.10.009

Cited by 8 publications

(15 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To demonstrate key features of the software and the utility of an object-oriented analysis approach, we began by reanalyzing a previously published desmosome dataset (7). Desmosomes are intercellular adhesive junctions responsible for maintaining mechanical integrity in epithelia and cardiac muscle (19).…”

Section: Resultsmentioning

confidence: 99%

“…To demonstrate key features of the software and the utility of an object-oriented analysis approach, we began by reanalyzing a previously published desmosome dataset (7).…”

Section: Object-based Analysis Of Punctate Structuresmentioning

confidence: 99%

“…FPM techniques come in many flavors (2), all of which are based upon the anisotropic nature of fluorescence absorption and emission (3). It is possible to study the organization of fluorescently labeled molecules microscopically by either modulating the orientation of the excitation polarization (4-7), separating the emission into polarized components (8, 9), or both (10, 11). We focus here on the excitation-resolved modality, which can be configured on a standard widefield microscope by simply placing a polarization modulating device in the illumination path.…”

Section: Introductionmentioning

confidence: 99%

“…Excitation-resolved FPM has provided novel insights into the architecture of cell-cell junctions (6, 7), the ordering of septin filaments in budding yeast (4, 5, 12), and the orientation of integrin traction forces (13), among others. However, full utilization of this technique has been hindered by the substantial image processing and mathematical analysis required to interpret the acquired data.…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

OOPS: Object-Oriented Polarization Software for analysis of fluorescence polarization microscopy images

Dean,

Nawara,

Albert

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Most essential cellular functions are performed by proteins assembled into larger complexes. Fluorescence Polarization Microscopy (FPM) is a powerful technique that goes beyond traditional imaging methods by allowing researchers to measure not only the localization of proteins within cells, but also their orientation or alignment within complexes or cellular structures. FPM can be easily integrated into standard widefield microscopes with the addition of a polarization modulator. However, the extensive image processing and analysis required to interpret the data have limited its widespread adoption. To overcome these challenges and enhance accessibility, we introduce OOPS (Object-Oriented Polarization Software), a MATLAB-based analysis tool tailored for FPM data. This work highlights the distinctive features of our software, which empower researchers to efficiently manage large datasets; detect and analyze individual structures; conduct population assessments based on morphology, intensity, and polarization-specific parameters; and create publication-quality visualizations, all within a user-friendly graphical interface. Importantly, OOPS is adaptable to various sample types, labeling techniques, and imaging setups, facilitating in-depth analysis of diverse polarization-sensitive samples. Here, we demonstrate the power and versatility of our approach by applying OOPS to FPM images of both punctate and filamentous structures. OOPS is freely available under the GNU GPL 3.0 license and can be downloaded athttps://github.com/Mattheyses-Lab/OOPS.

show abstract

Section: Resultsmentioning

confidence: 99%

“…To demonstrate key features of the software and the utility of an object-oriented analysis approach, we began by reanalyzing a previously published desmosome dataset (7).…”

Section: Object-based Analysis Of Punctate Structuresmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

OOPS: Object-Oriented Polarization Software for analysis of fluorescence polarization microscopy images

Dean,

Nawara,

Albert

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Through a variety of techniques, we and others have shown that cadherin ectodomains in mature desmosomes are organized into highly ordered assemblies [ 16–21 ], yet the relationship between cadherin architecture and desmosome function remains unclear. Given that the cadherins form the adhesive interface and provide a binding scaffold for plaque proteins, one possibility is that desmosomes rely on a precise arrangement of cadherins to supply adhesive strength and mechanical stability to the junction.…”

Section: Introductionmentioning

confidence: 99%

Dsg2 ectodomain organization increases throughout desmosome assembly

Dean,

Albert,

Nawara

et al. 2024

Cell Adhesion & Migration

Self Cite

View full text Add to dashboard Cite

Desmosomes are intercellular junctions that regulate mechanical integrity in epithelia and cardiac muscle. Dynamic desmosome remodeling is essential for wound healing and development, yet the mechanisms governing junction assembly remain elusive. While we and others have shown that cadherin ectodomains are highly organized, how this ordered architecture emerges during assembly is unknown. Using fluorescence polarization microscopy, we show that desmoglein 2 (Dsg2) ectodomain order gradually increases during 8 h of assembly, coinciding with increasing adhesive strength. In a scratch wound assay, we observed a similar increase in order in desmosomes assembling at the leading edge of migratory cells. Together, our findings indicate that cadherin organization is a hallmark of desmosome maturity and may play a role in conferring adhesive strength.

show abstract

Unraveling the geometry of complex protein organizations by polarized fluorescence imaging

Brasselet

2022

Biophysical Journal

View full text Add to dashboard Cite

Defining domain-specific orientational order in the desmosomal cadherins

Cited by 8 publications

References 52 publications

OOPS: Object-Oriented Polarization Software for analysis of fluorescence polarization microscopy images

OOPS: Object-Oriented Polarization Software for analysis of fluorescence polarization microscopy images

Dsg2 ectodomain organization increases throughout desmosome assembly

Unraveling the geometry of complex protein organizations by polarized fluorescence imaging

Contact Info

Product

Resources

About