Latest Publications

The Diverse Transformer (Trf) Protein Family in the Sea Urchin <em>Paracentrotus lividus</em> Acts through a Collaboration between Cellular and Humoral Immune Effector Arms

Fri, 07/02/2021 - 03:00

Int J Mol Sci. 2021 Jun 22;22(13):6639. doi: 10.3390/ijms22136639.

ABSTRACT

Sea urchins are long-living marine invertebrates with a complex innate immune system, which includes expanded families of immune receptors. A central immune gene family in sea urchins encodes the Transformer (Trf) proteins. The Trf family has been studied mainly in the purple sea urchin Strongylocentrotus purpuratus. Here, we explore this protein family in the Mediterranean Sea urchin Paracentrotus lividus. The PlTrf genes and predicted proteins are highly diverse and show a typical Trf size range and structure. Coelomocytes and cell-free coelomic fluid from P. lividus contain different PlTrf protein repertoires with a shared subset, that bind specifically to E. coli. Using FACS, we identified five different P. lividus coelomocyte sub-populations with cell surface PlTrf protein expression. The relative abundance of the PlTrf-positive cells increases sharply following immune challenge with E. coli, but not following challenge with LPS or the sea urchin pathogen, Vibrio penaeicida. Phagocytosis of E. coli by P. lividus phagocytes is mediated through the cell-free coelomic fluid and is inhibited by blocking PlTrf activity with anti-SpTrf antibodies. Together, our results suggest a collaboration between cellular and humoral PlTrf-mediated effector arms in the P. lividus specific immune response to pathogens.

PMID:34206148 | PMC:PMC8268236 | DOI:10.3390/ijms22136639

Categories: pubmed

Glyconectin Cell Adhesion Epitope, beta-d-GlcpNAc3S-(13)-alpha-l-Fucp, Is Involved in Blastulation of Lytechinus pictus Sea Urchin Embryos

Fri, 07/02/2021 - 03:00

Molecules. 2021 Jun 30;26(13):4012. doi: 10.3390/molecules26134012.

ABSTRACT

Glycans, as the most peripheral cell surface components, are the primary candidates to mediate the initial steps of cell recognition and adhesion via glycan-glycan binding. This molecular mechanism was quantitatively demonstrated by biochemical and biophysical measurements at the cellular and molecular level for the glyconectin 1 β-d-GlcpNAc3S-(1→3)-α-l-Fucp glycan structure (GN1). The use of adhesion blocking monoclonal antibody Block 2 that specifically recognize this epitope showed that, besides Porifera, human colon carcinoma also express this structure in the apical glycocalyx. Here we report that Block 2 selectively immune-precipitate a Mr 580 × 103 (g580) acidic non-glycosaminoglycan glycan from the total protein-free glycans of Lytechinus pictus sea urchin hatched blastula embryos. Immuno-fluorescence confocal light microscopy and immunogold electron microscopy localized the GN1 structure in the apical lamina glycocalyx attachments of ectodermal cells microvilli, and in the Golgi complex. Biochemical and immune-chemical analyses showed that the g580 glycan is carrying about 200 copies of the GN1 epitope. This highly polyvalent g580 glycan is one of the major components of the glycocalyx structure, maximally expressed at hatched blastula and gastrula. The involvement of g580 GN1 epitope in hatched blastula cell adhesion was demonstrated by: (1) enhancement of cell aggregation by g580 and sponge g200 glycans, (2) inhibition of cell reaggregation by Block 2, (3) dissociation of microvilli from the apical lamina matrix by the loss of its gel-like structure resulting in a change of the blastula embryonal form and consequent inhibition of gastrulation at saturating concentration of Block 2, and (4) aggregation of beads coated with the immune-purified g580 protein-free glycan. These results, together with the previous atomic force microscopy measurements of GN1 binding strength, indicated that this highly polyvalent and calcium ion dependent glycan-glycan binding can provide the force of 40 nanonewtons per single ectodermal cell association of microvilli with the apical lamina, and conservation of glycocalyx gel-like structure. This force can hold the weight of 160,000 cells in sea water, thus it is sufficient to establish, maintain and preserve blastula form after hatching, and prior to the complete formation of further stabilizing basal lamina.

PMID:34209220 | PMC:PMC8271808 | DOI:10.3390/molecules26134012

Categories: pubmed

Cnidarian-bilaterian comparison reveals the ancestral regulatory logic of the β-catenin dependent axial patterning

Wed, 06/30/2021 - 03:00

Nat Commun. 2021 Jun 29;12(1):4032. doi: 10.1038/s41467-021-24346-8.

ABSTRACT

In animals, body axis patterning is based on the concentration-dependent interpretation of graded morphogen signals, which enables correct positioning of the anatomical structures. The most ancient axis patterning system acting across animal phyla relies on β-catenin signaling, which directs gastrulation, and patterns the main body axis. However, within Bilateria, the patterning logic varies significantly between protostomes and deuterostomes. To deduce the ancestral principles of β-catenin-dependent axial patterning, we investigate the oral-aboral axis patterning in the sea anemone Nematostella-a member of the bilaterian sister group Cnidaria. Here we elucidate the regulatory logic by which more orally expressed β-catenin targets repress more aborally expressed β-catenin targets, and progressively restrict the initially global, maternally provided aboral identity. Similar regulatory logic of β-catenin-dependent patterning in Nematostella and deuterostomes suggests a common evolutionary origin of these processes and the equivalence of the cnidarian oral-aboral and the bilaterian posterior-anterior body axes.

PMID:34188050 | PMC:PMC8241978 | DOI:10.1038/s41467-021-24346-8

Categories: pubmed

Involvement of Huntingtin in Development and Ciliary Beating Regulation of Larvae of the Sea Urchin, <em>Hemicentrotus pulcherrimus</em>

Wed, 06/02/2021 - 03:00

Int J Mol Sci. 2021 May 12;22(10):5116. doi: 10.3390/ijms22105116.

ABSTRACT

The multiple functions of the wild type Huntington's disease protein of the sea urchin Hemicentrotus pulcherrimus (Hp-Htt) have been examined using the anti-Hp-Htt antibody (Ab) raised against synthetic oligopeptides. According to immunoblotting, Hp-Htt was detected as a single band at around the 350 kDa region at the swimming blastula stage to the prism larva stage. From the 2-arm pluteus stage (2aPL), however, an additional smaller band at the 165 kDa region appeared. Immunohistochemically, Hp-Htt was detected in the nuclei and the nearby cytoplasm of the ectodermal cells from the swimming blastula stage, and the blastocoelar cells from the mid-gastrula stage. The Ab-positive signal was converged to the ciliary band-associated strand (CBAS). There, it was accompanied by several CBAS-marker proteins in the cytoplasm, such as glutamate decarboxylase. Application of Hp-Htt morpholino (Hp-Htt-MO) has resulted in shortened larval arms, accompanied by decreased 5-bromo-2-deoxyuridin (BrdU) incorporation by the ectodermal cells of the larval arms. Hp-Htt-MO also resulted in lowered ciliary beating activity, accompanied by a disordered swirling pattern formation around the body. These Hp-Htt-MO-induced deficiencies took place after the onset of CBAS system formation at the larval arms. Thus, Hp-Htt is involved in cell proliferation and the ciliary beating pattern regulation signaling system in pluteus larvae.

PMID:34066037 | PMC:PMC8151597 | DOI:10.3390/ijms22105116

Categories: pubmed

When site matters: Metabolic and behavioural responses of adult sea urchins from different environments during long-term exposure to seawater acidification

Mon, 05/31/2021 - 03:00

Mar Environ Res. 2021 Jul;169:105372. doi: 10.1016/j.marenvres.2021.105372. Epub 2021 May 20.

ABSTRACT

CO2-driven ocean acidification (OA) affects many aspects of sea urchin biology. However, even in the same species, OA effects are often not univocal due to non-uniform exposure setups or different ecological history of the experimental specimens. In the present work, two groups of adult sea urchins Paracentrotus lividus from different environments (the Lagoon of Venice and a coastal area in the Northern Adriatic Sea) were exposed to OA in a long-term exposure. Animals were maintained for six months in both natural seawater (pHT 8.04) and end-of-the-century predicted condition (-0.4 units pH). Monthly, physiological (respiration rate, ammonia excretion, O:N ratio) and behavioural (righting, sheltering) endpoints were investigated. Both pH and time of exposure significantly influenced sea urchin responses, but differences between sites were highlighted, particularly in the first months. Under reduced pH, ammonia excretion increased and O:N decreased in coastal specimens. Righting and sheltering were impaired in coastal animals, whereas only righting decreased in lagoon ones. These findings suggested a higher adaptation ability in sea urchins from a more variable environment. Interestingly, as the exposure continued, animals from both sites were able to acclimate. Results revealed plasticity in the physiological and behavioural responses of sea urchins under future predicted OA conditions.

PMID:34058626 | DOI:10.1016/j.marenvres.2021.105372

Categories: pubmed