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ECB-ART-55233
Int J Mol Sci 2026 Jul 15;2714:. doi: 10.3390/ijms27146292.
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Proteomic Profiling of Density-Gradient Fractions Reveals Candidate Cell Surface Markers in Subpopulations of Starfish Asterias rubens Coelomocytes and Coelomic Epithelial Cells.

Shabelnikov SV, Sharlaimova NS, Bobkov DE, Mittenberg AG, Petukhova O.


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Cell surface markers are essential for lineage tracing and for understanding cellular mechanisms of regeneration, yet they remain largely unavailable for echinoderms. Using the starfish Asterias rubens as a model for physiological coelomocyte renewal, we combined Percoll density-gradient fractionation with label-free LC-MALDI proteomics to identify candidate surface proteins that distinguish subpopulations of coelomocytes and coelomic epithelium (CE) cells. However, LC-MALDI is prone to signal variability, sample depletion and nonlinear response. To address the quantitative limitations of LC-MALDI, we implemented a chi-square test with an empirically derived probability density function, which we validated on control mixtures, achieving high precision. Four fractions enriched in distinct cell types were analyzed: coelomocyte fractions 1 (roundish cells) and 4 (large agranulocytes that form cell nets) and CE fractions 1 (type 2 small CE cells) and 4 (ciliated cells), together with a weakly attached CE subpopulation enriched in small putative progenitor cells. LC-MALDI identified 714 protein groups, of which 160 showed significant differential abundance across fractions. Functional enrichment revealed actin cytoskeleton and integrin signaling in coelomocytes, and catenin-related terms in CE-W. Among the 101 predicted membrane proteins, 31 candidate cell surface markers were selected. These include integrins α8β1 and α9β1, three-finger proteins, scavenger receptors, and low-density lipoprotein receptor-related proteins, as well as metallopeptidases. This work provides the first proteome-scale set of candidate surface markers for starfish cell subpopulations, enabling future lineage-tracing studies to resolve the origin of coelomocytes from the coelomic epithelium.

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