Lectins identify distinct populations of coelomocytes in Strongylocentrotus purpuratus

Liao, Wen-Yun; Fugmann, Sebastian D.

doi:10.1371/journal.pone.0187987

Cited by 13 publications

(7 citation statements)

References 23 publications

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“…Coelomocytes are present throughout all adult tissues in S. purpuratus in addition to the CF and the fluid of the water vascular system (29, 36, 38) in which the red spherule cells can be observed as they circulate through the tube feet of S. purpuratus (LCS, personal observation). However, protein markers that identify all coelomocyte subsets are generally unknown and/or reagents are few (87). Consequently, it is difficult to demonstrate whether newly proliferated cells within tissues are coelomocytes, or, if they are coelomocytes, that they proliferated in the tissues in which they are observed.…”

Section: Resultsmentioning

confidence: 99%

The Axial Organ and the Pharynx Are Sites of Hematopoiesis in the Sea Urchin

et al. 2019

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Background: The location of coelomocyte proliferation in adult sea urchins is unknown and speculations since the early 1800s have been based on microanatomy and tracer uptake studies. In adult sea urchins ( Strongylocentrotus purpuratus ) with down-regulated immune systems, coelomocyte numbers increase in response to immune challenge, and whether some or all of these cells are newly proliferated is not known. The gene regulatory network that encodes transcription factors that control hematopoiesis in embryonic and larval sea urchins has not been investigated in adults. Hence, to identify the hematopoietic tissue in adult sea urchins, cell proliferation, expression of phagocyte specific genes, and expression of genes encoding transcription factors that function in the conserved regulatory network that controls hematopoiesis in embryonic and larval sea urchins were investigated for several tissues. Results: Cell proliferation was induced in adult sea urchins either by immune challenge through injection of heat-killed Vibrio diazotrophicus or by cell depletion through aspiration of coelomic fluid. In response to either of these stimuli, newly proliferated coelomocytes constitute only about 10% of the cells in the coelomic fluid. In tissues, newly proliferated cells and cells that express SpTransformer proteins (formerly Sp185/333) that are markers for phagocytes are present in the axial organ, gonad, pharynx, esophagus, and gut with no differences among tissues. The expression level of genes encoding transcription factors that regulate hematopoiesis show that both the axial organ and the pharynx have elevated expression compared to coelomocytes, esophagus, gut, and gonad. Similarly, an RNAseq dataset shows similar results for the axial organ and pharynx, but also suggests that the axial organ may be a site for removal and recycling of cells in the coelomic cavity. Conclusions: Results presented here are consistent with previous speculations that the axial organ may be a site of coelomocyte proliferation and that it may also be a center for cellular removal and recycling. A second site, the pharynx, may also have hematopoietic activity, a tissue that has been assumed to function only as part of the intestinal tract.

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Section: Resultsmentioning

confidence: 99%

The Axial Organ and the Pharynx Are Sites of Hematopoiesis in the Sea Urchin

et al. 2019

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“…However, fixation does not offer the isolation of live cells. The unavailability of antibodies targeting specific cell surface antigens of sea urchin coelomocytes limits the use of FACS based cell separation 24,25 . The results from a recent study indicate that lectin-based FACS could serve as a flow cytometry based strategy to purify distinct purple sea urchin (S. purpuratus) coelomocytes 25 .…”

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confidence: 99%

“…The unavailability of antibodies targeting specific cell surface antigens of sea urchin coelomocytes limits the use of FACS based cell separation 24,25 . The results from a recent study indicate that lectin-based FACS could serve as a flow cytometry based strategy to purify distinct purple sea urchin (S. purpuratus) coelomocytes 25 . Lectins are known to target glycosylation patterns presented by individual cell types [26][27][28][29] and this interaction could thus be utilised to define different immune cell populations.…”

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confidence: 99%

“…The glycosylation pattern usually reflects different cellular states and therefore, may alter lectin interaction. For example, Solanum tuberosum lectin (STL) fails to bind phagocytic cells of sea urchins settled onto glass slides, but binds successfully when the cells are in solution 25 . The RSCs of sea urchins are shown to possess a dynamic array of cellular states 31 .…”

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confidence: 99%

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Autofluorescence mediated red spherulocyte sorting provides insights into the source of spinochromes in sea urchins

Hira

Wolfson

Andersen

et al. 2020

Sci Rep

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Red spherule cells (RSCs) are considered one of the prime immune cells of sea urchins, but their detailed biological role during immune responses is not well elucidated. Lack of pure populations accounts for one of the major challenges of studying these cells. In this study, we have demonstrated that live RSCs exhibit strong, multi-colour autofluorescence distinct from other coelomocytes, and with the help of fluorescence-activated cell sorting (FACS), a pure population of live RSCs was successfully separated from other coelomocytes in the green sea urchin, Strongylocentrotus droebachiensis. This newly developed RSCs isolation method has allowed profiling of the naphthoquinone content in these cells. With the use of ultra high-performance liquid chromatography, UV absorption spectra, and highresolution tandem mass spectrometry, it was possible to identify sulphated derivatives of spinochrome C, D, E and spinochrome dimers, which suggests that the RSCs may play an important biological role in the biogenesis of naphthoquinone compounds and regulating their bioactivity.

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“…Direct comparison of echinoid coelomocyte types identified by FC with those of Asteroidea is difficult given the known variability of coelomocyte types among echinoderm classes (7); (22) and by the diversity of protocols, for instance, the specific lectins-labelling method used by Liao and collaborators (2017) (26).…”

Section: Coelomocyte Fractionation Using Flow Cytometrymentioning

confidence: 99%

A combined characterization of coelomic fluid cell types in the spiny starfishMarthasterias glacialis– inputs from flow cytometry and imaging

Oliveira

Guatelli

Martı́nez

et al. 2020

Preprint

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Coelomocytes is a generic name for a collection of cellular morphotypes, present in many coelomate animals, that has been reported as highly variable across echinoderm classes. The roles attributed to the major types of the free circulating cells present in the coelomic fluid of echinoderms include immune response, phagocytic digestion and clotting. The main aim of the present study is the thorough characterization of coelomocytes present in the coelomic fluid of Marthasterias glacialis (class Asteroidea) through the combined use of flow cytometry (FC) and fluorescence plus transmission electron microscopy. Two coelomocyte populations (here named P1 and P2) were identified by flow cytometry and subsequently studied in terms of abundance, morphology, ultrastructure, cell viability and cell cycle profiles. Ultrastructurally, P2 diploid cells showed two main morphotypes, similar to phagocytes and vertebrate thrombocytes, whereas the small P1 haploid cellular population was characterized by a low mitotic activity, relatively undifferentiated cytotype and a high nucleus/cytoplasm ratio. These last cells resemble stem-cell types present in other animals. P1 and P2 cells differ also in cell viability and cell cycle profiles. Additionally, two other morphotypes were only detected by fluorescence microscopy and a third one when using a combined microscopy/FC approach.

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Lectins identify distinct populations of coelomocytes in Strongylocentrotus purpuratus

Cited by 13 publications

References 23 publications

The Axial Organ and the Pharynx Are Sites of Hematopoiesis in the Sea Urchin

The Axial Organ and the Pharynx Are Sites of Hematopoiesis in the Sea Urchin

Autofluorescence mediated red spherulocyte sorting provides insights into the source of spinochromes in sea urchins

A combined characterization of coelomic fluid cell types in the spiny starfishMarthasterias glacialis– inputs from flow cytometry and imaging

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