A systematic fluorescently tagged protein toolkit for dissecting apical extracellular matrix patterning in C. elegans
- 1Department of Molecular Cell and Developmental Biology, University of California Santa Cruz, Santa Cruz, CA 95064
- 2Department of Cell and Developmental Biology, School of Biological Sciences, University of California San Diego, La Jolla, CA 92093
Abstract
Barrier epithelia are shielded from the external environment by their apical extracellular matrices (aECMs). The molecular complexity of aECMs has challenged understanding of their organization in vivo. To define the molecular architecture of a model aECM we generated a toolkit of 105 fluorescently tagged aECM components using gene editing in C. elegans, focusing on proteins secreted by the epidermis to form the collagen-rich cuticle. We developed efficient pipelines for modular protein tagging and rapid fluorophore swapping. Most tagged collagens were functional and exhibited exquisitely specific patterning across stages, cell types, and matrix substructures. We define multiple reference markers for key substructures including the little-understood cortical layer, as well as the helical crossed fiber arrays that function as a hydrostatic skeleton to maintain organismal shape. We further tagged 30 members of key aECM protein classes including proteases, protease inhibitors, and lipid transporters. Our standardized markers will allow dissection of the mechanistic basis of aECM spatiotemporal patterning in vivo.
Barrier epithelia are protected by apical extracellular matrices (aECMs), but their complex organization remains poorly understood in vivo. This study generates a toolkit of 105 fluorescently tagged aECM components and defines reference markers for key substructures, enabling analysis of spatiotemporal extracellular matrix patterning.
Related
- Dissecting the pro-neurogenic effects of monoaminergic medications used to treat depression: a systematic review and meta-analysis
- An atypical cysteine-type protein domain regulates cuticular extracellular matrix assembly in nematodes
- Extracellular Vesicle Protein Cargo Reveals Potential Long-Term Biological Impact of Long COVID
- A TurboID-based proximity labelling method for detecting extracellular protein interactions in plants
- Cellular Prion Protein Coupling to ADAM‐mediated Release of Small Extracellular Vesicles: Pathological Implications in Prostate Cancer Cells
BibTeX
@article{ragle2026systematic,
title = {{A systematic fluorescently tagged protein toolkit for dissecting apical extracellular matrix patterning in C. elegans}},
author = {James Matthew Ragle and Murugesan Pooranachithra and Guinevere E. Ashley and Emma Cadena and Batia Blank and Katelyn Kang and Cincy Chen and Aditree R. Bhowmick and Soraya Hawk Mercado and Tabatha E. Wells and John C Clancy and Andrew D. Chisholm and Jordan D. Ward},
journal = {PLOS Biology},
year = {2026},
month = oct,
doi = {10.1371/journal.pbio.3004059},
url = {https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004059},
note = {CC BY},
}
RIS
TY - JOUR
TI - A systematic fluorescently tagged protein toolkit for dissecting apical extracellular matrix patterning in C. elegans
AU - James Matthew Ragle
AU - Murugesan Pooranachithra
AU - Guinevere E. Ashley
AU - Emma Cadena
AU - Batia Blank
AU - Katelyn Kang
AU - Cincy Chen
AU - Aditree R. Bhowmick
AU - Soraya Hawk Mercado
AU - Tabatha E. Wells
AU - John C Clancy
AU - Andrew D. Chisholm
AU - Jordan D. Ward
PY - 2026
DA - 2026/10/14
JO - PLOS Biology
DO - 10.1371/journal.pbio.3004059
UR - https://journals.plos.org/plosbiology/article?id=10.1371/journal.pbio.3004059
AB - Barrier epithelia are shielded from the external environment by their apical extracellular matrices (aECMs). The molecular complexity of aECMs has challenged understanding of their organization in vivo. To define the molecular architecture of a model aECM we generated a toolkit of 105 fluorescently tagged aECM components using gene editing in C. elegans, focusing on proteins secreted by the epidermis to form the collagen-rich cuticle. We developed efficient pipelines for modular protein tagging and rapid fluorophore swapping. Most tagged collagens were functional and exhibited exquisitely specific patterning across stages, cell types, and matrix substructures. We define multiple reference markers for key substructures including the little-understood cortical layer, as well as the helical crossed fiber arrays that function as a hydrostatic skeleton to maintain organismal shape. We further tagged 30 members of key aECM protein classes including proteases, protease inhibitors, and lipid transporters. Our standardized markers will allow dissection of the mechanistic basis of aECM spatiotemporal patterning in vivo. Barrier epithelia are protected by apical extracellular matrices (aECMs), but their complex organization remains poorly understood in vivo. This study generates a toolkit of 105 fluorescently tagged aECM components and defines reference markers for key substructures, enabling analysis of spatiotemporal extracellular matrix patterning.
SN - 1545-7885
SN - 1544-9173
ER -