Gene expression and regulation of molecules involved in pharynx inflammatory response induced by LPS in Ciona intestinalis

In the ascidian Ciona intestinalis, the pharynx (hemopoietic organ) connects the external environment to the gastrointestinal system for two main activities, respiration and food collection, potentially exposing the ascidian to high concentrations of pathogenic microorganisms. Recently, evidence in C. intestinalis has indicated that the pharynx is involved in the inflammatory reaction induced by lipopolysaccharide (LPS) injection into the body wall. Immune-related genes such as cytokines, galectins, pro-PO, CAP are expressed in pharynx hemocytes and are up-regulated by the inflammatory agent LPS. Studies of the expression pattern of the immune gene clearly show that in C. intestinalis, as in vertebrates, immune gene expression can be regulated through Alternative Polyadenylation (APA) Mechanism and GAIT element al 3\u2019UTR

Molecular characterisation, evolution and expression analysis of g-type lysozymes in Ciona intestinalis

Lysozyme is an important defense molecule of the innate immune system. Known for its bactericidal properties, lysozyme catalyzes the hydrolysis of b-(1,4)-glycosidic bonds between the N-acetyl glucosamine and N-acetyl muramic acid in the peptidoglycan layer of bacterial cell walls. In this study, the complete coding sequence of four g-type lysozymes were identified in Ciona intestinalis. Phylogenetic analysis and modelling supported the hypothesis of a close relationship with the vertebrate g-type lysozymes suggesting that the C. intestinalis g-type lysozyme genes (CiLys-g1, Cilys-g2, CiLys-g3, CiLys-g4) share a common ancestor in the chordate lineage. Protein motif searches indicated that C. intestinalis gtype lysozymes contain a GEWL domain with a GXXQ signature, typical of goose lysozymes. Quantitative Real-Time PCR analysis results showed that transcripts are expressed in various tissues from C. intestinalis. In order to determine the involvement of C. intestinalis g-type lysozymes in immunity, their expression was analyzed in the pharynx, showing that transcripts were significantly up-regulated in response to a challenge with lipopolysaccharide (LPS). These data support the view that CiLys g-type are molecules with potential for immune defense system against bacterial infection

Bottom-Up Control of Macrobenthic Communities in a Guanotrophic Coastal System

Soft bottom macrobenthic communities were studied seasonally in three coastal ponds (Marinello ponds, Italy) at increasing distances from a gull (Larus michahellis) colony to in- vestigate the effect of seabird-induced eutrophication (i.e. guanotrophication) on macro- benthic fauna.We hypothesized that enhanced nutrient concentration and organic load caused by guano input significantly alter the trophic and sedimentological condition of ponds, affecting benthic fauna through a bottom-up control. The influence of a set of envi- ronmental features on macrobenthic assemblages was also tested. Overall, the lowest macrobenthic abundances and functional group diversity were found in deeper sites, espe- cially in the pond characterised by severe guanotrophication, where the higher disturbance resulted in a decline in suspension feeders and carnivores in favour of deposit feeders. An increase in opportunistic/tolerant taxa (e.g. chironomid larvae and paraonids) and totally azoic sediments were also found as an effect of the harshest environmental conditions, re- sulting in a very poor ecological status. We conclude that macrobenthic assemblages of the Marinello coastal system display high spatial variability due to a synergistic effect of trophic status and the geomorphological features of the ponds. Themacrobenthic response to gua- notrophication, which was a clear decrease in abundance, diversity and trophic functional groups, was associated with the typical response to severe eutrophication, magnified by the geomorphological features

The Atlantic blue crab Callinectes sapidus spreading in the Tyrrhenian sea: evidence of an established population in the Stagnone di Marsala (Sicily, southern Italy)

The blue crab Callinectes sapidus is a portunid brachyuran native of the Atlantic coasts of America. In the last decades, the species has greatly expanded its range in invaded areas, currently including the Atlantic coasts of Morocco, the Mediterranean Sea, the Black Sea as well as the eastern Atlantic coasts of Europe from Portugal to France, Belgium, and Germany. In the present study, several specimens of C. sapidus were collected in May, June and July 2021 in the Stagnone di Marsala, a semi-enclosed basin located in the north-western coast of Sicily (southern Italy, Mediterranean Sea) characterized by low hydrodynamics and water exchange with the adjacent open sea and high salinity conditions. Juveniles at diff erent instar stages and ovigerous females were repeatedly captured, suggesting the presence of an established population of the blue crab, despite the peculiar environmental conditions characterizing the basin. These fi ndings are discussed in the context of the current expansion of the species along the African coasts of the Mediterranean Sea

A new Leucoagaricus species of section Piloselli (Agaricales, Agaricaceae) from Spain

The new species Leucoagaricus variicolor is described from a public park in Zaragoza, Spain, based on both morphological and molecular characters. Illustrations of fresh basidiomata in situ and of the main macro- and micromorphological features are added. Leucoagaricus variicolor belongs to section Piloselli and is compared with similar taxa

Ciona intestinalis galectin (CiLgals-a and CiLgals-b) genes are differentially expressed in endostyle zones and challenged by LPS

Immunohistochemical and in situ hybridization assays were performed to answer the question whether the endostyle, that is the initial gastro-intestinal trait of Ciona intestinalis pharynx, is involved in galectin (CiLgals-a and CiLgals-b) production during the pharynx inflammatory response to LPS inoculation. Specific anti-CiLgal-a and anti-CiLgals-b antibodies, and oligonucleotide probes, that mark inflammatory hemocytes inside the pharynx vessels and vessel epithelium as shown by a previous paper, were assayed on endostyle histological sections. For the first time, we show that galectins are produced by endostyle zones, and both CiLgals-a and eb genes are upregulated by LPS. CiLgals-a and CiLgals-b are constitutively expressed in the endostyle zone 2 and 3, respectively, both genes are upregulated by LPS in the zone 2, and CiLgals-b in the zone 3 and 4. The antibody-reacting material contained in intracellular and extracellular large vesicles suggest an unexpected vesicle-dependent transporting mechanism of galectins not provided with signal peptide. Differential expression and gene upregulation in not-treated and LPStreated specimens, support the role of endostyle galectins both in filter feeding and defense responses

In the ovary of Ciona intestinalis (Type A), immune-related galectin and phenoloxidase genes are differentially expressed by the follicle accessory cells

Riboprobes (in situ hybridization) and antibodies (immunohistochemistry), previously used to show the upregulation of Ciona intestinalis (Type A) galectins (CiLgals-a, CiLgals-b) and phenoloxidase (CinPO2) immune-related genes, were tested on histological sections of the ovary. The ovarian follicles are composed of oocytes encased by follicular cells (FCs) and test cells (TCs). Results show the transcription upregulation of both CiLgals and CinPO2 genes in the vitellogenic FCs, conversely distinct cytolocalization of the proteins are shown. At vitellogenic stage, the CiLgals are localized in the FCs, in the oocyte cytoplasm, and close to the germinal vesicle (GV), whereas the CinPO2 was never identified in the FCs. In a presumptive advanced phase and at the post-vitellogenic stage the TCs appear to be labelled by the CinPO2 riboprobe, and the protein identified by the antibody suggesting an mRNA transcytosis process from FCs. At post-vitellogenic stage the CiLgals mainly enrich the GV nucleoplasm, whereas the CinPO2 is contained in TCs and in the ooplasm but never found in the GV. This finding sheds new light on a former paper in which TCs were reported to be the only CinPO2-producing cells in the ovarian follicle. Finally, CiLgals and CinPO2 genes transcription and proteins production seem to be associated with accessory cells during their differentiation from vitellogenic to post-vitellogenic stage. The present findings promote further research on the early upregulation of immune-related genes, and the potential multifunctional role of the produced proteins. In addition further insight on the accessory cells involvement in ascidian oogenesis are reported

Isolation and expression of a novel MBL-like collectin cDNA enhanced by LPS injection in the body wall of the ascidian Ciona intestinalis

Collectins are a family of calcium-dependent lectins that are characterized by their collagen-like domains. Considerable interest has been focused on this class of proteins because of their ability to interact with components of the complement system activating a cascade of events responsible for the activation of the innate immune system. A differential screening between LPS-challenged and na\uefve Ciona intestinalis has been performed allowing the isolation of a full length cDNA encoding for a 221 AA protein. In silico analysis has shown that this polypeptide displays protein domains with similarities to mannose-binding lectins. A phylogenetic analysis suggested that C. intestinalis MBL has evolved early as a prototype of vertebrate MBL. Real-time PCR assay demonstrated that this gene is strongly activated after LPS injection in the tunica. In situ hybridization performed in LPS-induced animals has shown that this gene is expressed in granular amoebocytes and large granules hemocytes in the inflamed body wall tissue. Finally, an antimicrobial activity of the C. intestinalis MBL has been demonstrate