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The polian vesicle and coelom of sea cucumber Apostichopus japonicus were full of coelomic fluid in which many types of coelomocytes with different functions were suspended. Our previous work has indicated the differences of coelomocytes between two sites mainly in subtype proportion, non-specific immune enzymes activities and several immune-related genes expression levels in healthy A. japonicus. However, the functional similarities and differences of coelomic fluid in two sites including the coelom and polian vesicle after pathogenic infection still remain unclear. Here, we investigated the changes of the total coelomocyte density (TCD) and differential coelomocyte density (DCD) after pathogen infection by Vibrio splendidus in coelom and polian vesicle. After infected by V. splendidus, the TCD in the coelom and polian vesicle rapidly declined at 12 h, and then the TCD in the coelom showed a stably ascending trend, while the TCD in the polian vesicle reached a peak at 24 h post infection (hpi), and then showed a continuously decline trend from 24 hpi to 72 hpi followed by a slow elevation until recovering the normal level from 72 hpi to 96 hpi. Then the activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD) were determined to evaluate the response of cell-free coelomic fluid to V. splendidus infection. The activities of ACP, AKP and CAT showed similar trends in the coelom and polian vesicle. The SOD activity significantly increased in the polian vesicle, whereas it exhibited a decreasing trend in the coelom. Finally, the expression profiles of nine immune-related genes including Aj-MyD88, Aj-IRAK4, Aj-i-Lys, Aj-Rel, Aj-p50, Aj-DMBT1, Aj-CDC, Aj-Rrp15 and Aj-Fibrinogen C were detected after V. splendidus challenge. The results suggested all the detected genes were significantly up-regulated both in the coelom and polian vesicle, and the expression levels of these genes in two sites shared similar trends except Aj-MyD88 and Aj-DMBT1. This research provides a new insight into the differentially immune roles of coelomic fluid and coelomocytes in polian vesicle and coelom response to bacterial infections and supplements comprehensive resources for better understanding the innate immune response of A. japonicus.

Copyright of Journal of Dalian Ocean University is the property of Journal of Dalian Ocean University Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Coelomic fluid surrounds the internal organs of asteroid echinoderms (asteroids, otherwise known as sea stars or starfish) and plays an essential role in the immune system, as well as in the transport of respiratory gases, nutrients, waste products, and reproductive mediators. Due to its importance in physiology and accessibility for nonlethal diagnostic sampling, coelomic fluid of asteroids provides an excellent sample matrix for health evaluations and can be particularly useful in disease and mortality investigations. This is especially important in light of recent increases in the number of affected individuals and species, larger geographic scope, and increased observed frequency of sea star wasting events compared with historic accounts of wasting. This review summarizes the current knowledge about coelomocytes, the effector cell of the asteroid immune system; coelomic fluid electrolytes, osmolality, acid–base status and respiratory gases, and microbiota; and genomic, transcriptomic, and proteomic investigations of coelomic fluid. The utility of coelomic fluid analysis for assessing stressor responses, diseases, and mortality investigations is considered with knowledge gaps and future directions identified. This complex body fluid provides an exciting opportunity to increase our understanding of this unique and ecologically important group of animals. [ABSTRACT FROM AUTHOR]

Marine animals in the wild are often difficult to access, so they are studied in captivity. However, the implicit assumption that physiological processes of animals in artificial environments are not different from those in the wild has rarely been tested. Here, we investigate the extent to which an animal is impacted by captivity by comparing global gene expression in wild and captive crown‐of‐thorns starfish (COTS). In a preliminary analysis, we compared transcriptomes of three external tissues obtained from multiple wild COTS with a single captive COTS maintained in aquaria for at least 1 week. On average, an astonishingly large 24% of the coding sequences in the genome were differentially expressed. This led us to conduct a replicated experiment to test more comprehensively the impact of captivity on gene expression. Specifically, a comparison of 13 wild with 8 captive COTS coelomocyte transcriptomes revealed significant differences in the expression of 20% of coding sequences. Coelomocyte transcriptomes in captive COTS remain different from those in wild COTS for more than 30 days and show no indication of reverting back to a wild state (i.e. no evidence of acclimation). Genes upregulated in captivity include those involved in oxidative stress and energy metabolism, whereas genes downregulated are involved in cell signalling. These changes in gene expression indicate that being translocated and maintained in captivity has a marked impact on the physiology and health of these echinoderms. This study suggests that caution should be exercised when extrapolating results from captive aquatic invertebrates to their wild counterparts. [ABSTRACT FROM AUTHOR]

• Pathological changes showed a chlorine substitution-dependent toxicity. • Highly chlorinated PCBs had lower accumulation capacities to earthworms. • Lowly chlorinated PCBs were less toxic to Eisenia fetida. Polychlorinated biphenyls (PCBs) with different chlorine substitution patterns often coexist in e-waste-processing sites. However, the single and combined toxicity of PCBs to soil organisms and the influence of chlorine substitution patterns remain largely unknown. Herein, we evaluated the distinct in vivo toxicity of PCB28 (a trichlorinated PCB), PCB52 (a tetrachlorinated PCB), PCB101 (a pentachlorinated PCB), and their mixture to earthworm Eisenia fetida in soil, and looked into the underlining mechanisms in an in vitro test using coelomocytes. After a 28-days exposure, all PCBs (up to 10 mg/kg) were not fatal to earthworms, but could induce intestinal histopathological changes and microbial community alterations in the drilosphere system, along with a significant weight loss. Notably, pentachlorinated PCBs with a low bioaccumulation ability showed greater inhibitory effects on the growth of earthworm than lowly chlorinated PCBs, suggesting that bioaccumulation was not the main determinant of chlorine substitution-dependent toxicity. Furthermore, in vitro assays showed that the highly chlorinated PCBs induced a high-percentage apoptosis of eleocytes in the coelomocytes and significantly activated antioxidant enzymes, indicating that the distinct cellular vulnerability to lowly/highly chlorinated PCBs was the main contributor to the PCBs toxicity. These findings emphasize the specific advantage of using earthworms in the control of lowly chlorinated PCBs in soil due to their high tolerance and accumulation ability. [Display omitted] [ABSTRACT FROM AUTHOR]

Earthworms (Eisenia fetida) were exposed to individual and binary mixture of imidacloprid (IMI) and dinotefuran (DIN) at 0.05 and 0.5mg/kg for 28 days to investigate their bioaccumulation, transformation and toxicity. IMI was more easily absorbed by earthworms than DIN, and worms didn't accumulate or generate toxic metabolites. The obvious accumulation of neonicotinoids during later period caused significant neural dysfunction, especially when exposed to high-concentration IMI. Meanwhile, oxidative stress indicated by decreased SOD/CAT activity (33.2%-68.1%) and increased MDA (38.4%-55.0%) was induced by binary exposure with high-concentration IMI. By contrast, coelomocytes responded earlier and more strongly than oxidative responses. Coelomocytes' viability and mitochondrial membrane potential were inhibited (23.6%-91.7%) mainly by IMI and binary exposure. Coelomocytes' lactate dehydrogenase activity exerted a fluctuating pattern, suggesting irregular disturbance on cellular functions. This study highlights the role of coelomocytes and the need to consider binary/multiple scenarios and transformation of neonicotinoids in their risk assessment to earthworms., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Development of multicellular organisms requires well-orchestrated interplay between cell-intrinsic transcription factors and cell-cell signaling. One set of highly conserved transcription factors that plays diverse roles in development is the SoxC group. C. elegans contains a sole SoxC protein, SEM-2. SEM-2 is essential for embryonic development, and for specifying the sex myoblast (SM) fate in the postembryonic mesoderm, the M lineage. We have identified a novel partial loss-of-function sem-2 allele that has a proline to serine change in the C-terminal tail of the highly conserved DNA-binding domain. Detailed analyses of mutant animals harboring this point mutation uncovered new functions of SEM-2 in the M lineage. First, SEM-2 functions antagonistically with LET-381, the sole C. elegans FoxF/C forkhead transcription factor, to regulate dorsoventral patterning of the M lineage. Second, in addition to specifying the SM fate, SEM-2 is essential for the proliferation and diversification of the SM lineage. Finally, SEM-2 appears to directly regulate the expression of hlh-8 , which encodes a basic helix-loop-helix Twist transcription factor and plays critical roles in proper patterning of the M lineage. Our data, along with previous studies, suggest an evolutionarily conserved relationship between SoxC and Twist proteins. Furthermore, our work identified new interactions in the gene regulatory network (GRN) underlying C. elegans postembryonic development and adds to the general understanding of the structure-function relationship of SoxC proteins.

A representative echinoderm, the sea cucumber Apostichopus japonicus has a special regeneration mechanism. The sea cucumber has no specific immune tissues or organs. Coelomocytes of sea cucumbers are involved in their cellular and humoral immunity. In this study, expression profiles of the main immune-related factors in sea cucumber coelomocytes were detected during coelomocyte regeneration after evisceration in A. japonicus. Immune-related factors Cu/Zn superoxide dismutase (Cu/Zn SOD), Catalase (CAT), C-type lysozyme (C-LYZ), Cathepsin D (CTSD), Melanotransferrin (MTF), Toll-like receptor (TLR), C-type lectin (C-LCT), Complement 3 (C3), Myeloid differentiation factor 88 (MYD88), Nuclear factor kappa-B (NF-κB), NF-κB inhibitor (IKB), TNF receptor-associated factor 6 (TRAF6), Peroxiredoxins (PRX), Nitric oxide synthase(NOS), Caspase-2 (CASP-2), Phenoloxidase (PO), and Glutathione Peroxidase(GPX), Heat shock protein 70 (HSP70) were detected by real-time fluorescent quantitative PCR at different time points during regeneration. The main immune-related genes in sea cucumber coelomocytes were significantly differentially expressed after evisceration, and an upregulation was observed for the majority of the considered genes. In summary, the discharge of viscera had a significant effect on expression of immune-related genes of sea cucumber coelomocytes. The expression level of each gene had a certain correlation with the sea cucumber regeneration process. The results provide reference data for the immune response of coelomocytes during regeneration.

A representative echinoderm, the sea cucumber Apostichopus japonicus has a special regeneration mechanism. The sea cucumber has no specific immune tissues or organs. Coelomocytes of sea cucumbers are involved in their cellular and humoral immunity. In this study, expression profiles of the main immune-related factors in sea cucumber coelomocytes were detected during coelomocyte regeneration after evisceration in A. japonicus. Immune-related factors Cu/Zn superoxide dismutase (Cu/Zn SOD), Catalase (CAT), C-type lysozyme (C-LYZ), Cathepsin D (CTSD), Melanotransferrin (MTF), Toll-like receptor (TLR), C-type lectin (C-LCT), Complement 3 (C3), Myeloid differentiation factor 88 (MYD88), Nuclear factor kappa-B (NF-κB), NF-κB inhibitor (IKB), TNF receptor-associated factor 6 (TRAF6), Peroxiredoxins (PRX), Nitric oxide synthase(NOS), Caspase-2 (CASP-2), Phenoloxidase (PO), and Glutathione Peroxidase(GPX), Heat shock protein 70 (HSP70) were detected by real-time fluorescent quantitative PCR at different time points during regeneration. The main immune-related genes in sea cucumber coelomocytes were significantly differentially expressed after evisceration, and an upregulation was observed for the majority of the considered genes. In summary, the discharge of viscera had a significant effect on expression of immune-related genes of sea cucumber coelomocytes. The expression level of each gene had a certain correlation with the sea cucumber regeneration process. The results provide reference data for the immune response of coelomocytes during regeneration. [ABSTRACT FROM AUTHOR]

To characterize the immune functions of the coelomic fluid of echiuran worms (Urechis unicinctus), their cellular composition and biochemical index were initially analyzed. Meanwhile, the phagocytic activity of coelomocytes and the impact of pathogenic infection immuno-related enzymes were explored. Lastly, coelomocyte density and humoral immune factor levels were detected under pathogenic conditions. The results revealed three major coelomocyte types in the coelomic fluid, comprising granular cells, hyaline cells and thrombocytes. Granulosa cells exhibited ability to adsorb and phagocytose with bacteria, whereas hyaline cells and thrombocytes manifested no adsorption and phagocytosis abilities. Additionally, numerous immune factors (such as albumin, alanine aminotransferase, and alkaline phosphatase) were detected in the coelomic fluid. Following pathogenic stimulation, the coelomocyte density and level of humoral immune factors (SOD, LSZ, ACP, and AKP) were changed. Collectively, the results suggested that humoral immune factors and coelomocytes were sensitive to environmental factors. Taken together, our study indicated that coelomocytes, in conjunction with immune factors in the coelomic fluid, formed an immune barrier in U. unicinctus. • The prime coelomocytes types were granular cells, hyaline cells, and thrombocytes. • Granulosa cells exhibited adsorption and phagocytosis abilities with bacteria. • The humoral immune factors levels were altered after pathogen stimulation. • The coelomocytes and immune factors support the defense system of U. unicinctus. [ABSTRACT FROM AUTHOR]

Lysosomes, the major membrane-bound degradative organelles, have a multitude of functions in eukaryotic cells. Lysosomes are the terminal compartments in the endocytic pathway, though they display highly dynamic behaviors, fusing with each other and with late endosomes in the endocytic pathway, and with the plasma membrane during regulated exocytosis and for wound repair. After fusing with late endosomes, lysosomes are reformed from the resulting hybrid organelles through a process that involves budding of a nascent lysosome, extension of the nascent lysosome from the hybrid organelle, while remaining connected by a membrane bridge, and scission of the membrane bridge to release the newly formed lysosome. The newly formed lysosomes undergo cycles of homotypic fusion and fission reactions to form mature lysosomes. In this study, we used a forward genetic screen in Caenorhabditis elegans to identify six regulators of lysosome biology. We show that these proteins function in different steps of lysosome biology, regulating lysosome formation, lysosome fusion, and lysosome degradation.

The immune system of the sea urchin species Paracentrotus lividus is highly complex and, as yet, poorly understood. P. lividus coelomocytes mediate immune response through phagocytosis and encapsulation of non-self particles, in addition to the production of antimicrobial molecules. Despite this understanding, details of exactly how these processes occur and the mechanisms which drive them are still in need of clarification. In this study, we show how the bacterial lipopolysaccharides (LPS) is able to induce a stress response which increases the levels of the heat shock proteins HSP70 and HSP90 only a few hours after treatment. This study also shows that LPS treatment increases the expression of the β-thymosin-derivated protein paracentrin, the precursor of antimicrobial peptides. • Immune response in Paracentrotus lividus after stimulation with LPS. • Echinoderm immune response and HSP90 and HSP70 expression. • β-Thymosin modulation following LPS treatment. [ABSTRACT FROM AUTHOR]

Copyright of Journal of Agricultural Science & Technology (1008-0864) is the property of Journal of Agricultural Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract Sea cucumber Apostichopus japonicus rely on the efficient innate immune mechanisms against invaders, in which the consumption and regeneration of coelomocytes take place at the same time. In the present study, histological features of putative hematopoietic tissues (HPTs) including the rete mirabile, the respiratory tree, the polian vesicle and the coelomic epithelium were characterized. The distribution of transcription factor GATA1 in coelomocytes and putative HPTs was examined by immunohistochemistry. In addition, cell proliferation using EdU labeling and coelomocyte distribution in different tissues using monoclonal antibody labeling were analyzed to further confirm the HPTs. The results showed that two homologs of GATA1 were detected with molecular weight of 43 and 90 kDa in coelomocytes, rete mirabile, respiratory tree and polian vesicle, whereas no signals were detected in the coelomic epithelium. A few cells were detected to be EdU-positive for coelomocytes, which accounted for approximately 9.5%. In the rete mirabile and the respiratory tree, the EdU signals were strong in cells of the tube wall. In the polian vesicle, numerous EdU-positive cells were detected in the cyst wall. In the coelomic epithelium, little EdU signaling was detected. Immunohistochemistry analysis by mAb 3F6 against A. japonicus coelomocytes showed that positive signals were observed in the tube wall of the rete mirabile, respiratory tree, cyst wall of the polian vesicle and in the coelomocyte antrum of coelomic epithelium. These results suggest that the rete mirabile, respiratory tree and polian vesicle are the HPTs of A. japonicus. Highlights • Rete mirabile, respiratory tree, and polian vesicle were suggested as HPTs. • Two homologs of transcription factor GATA1 were detected in the HPTs. • EdU signals were strong in coelomocytes and HPTs. [ABSTRACT FROM AUTHOR]

Titanium dioxide nanoparticles (TiO2 NPs) are manufactured worldwide. Once they arrive in the soil environment, they can endanger living organisms. Hence, monitoring and assessing the effects of these nanoparticles is required. We focus on the Eisenia andrei earthworm immune cells exposed to sublethal concentrations of TiO2 NPs (1, 10, and 100 µg/mL) for 2, 6, and 24 h. TiO2 NPs at all concentrations did not affect cell viability. Further, TiO2 NPs did not cause changes in reactive oxygen species (ROS) production, malondialdehyde (MDA) production, and phagocytic activity. Similarly, they did not elicit DNA damage. Overall, we did not detect any toxic effects of TiO2 NPs at the cellular level. At the gene expression level, slight changes were detected. Metallothionein, fetidin/lysenin, lumbricin and MEK kinase I were upregulated in coelomocytes after exposure to 10 µg/mL TiO2 NPs for 6 h. Antioxidant enzyme expression was similar in exposed and control cells. TiO2 NPs were detected on coelomocyte membranes. However, our results do not show any strong effects of these nanoparticles on coelomocytes at both the cellular and molecular levels.

The concurrent existence of cadmium (Cd) and ciprofloxacin (CIP) in agricultural soils is very common, but presents a challenge to soil organisms. As more attention has been paid to the effect of toxic metals on the migration of antibiotic resistance genes, the critical role of the gut microbiota in CIP-modifying Cd toxicity in earthworms remains unclear. In this study, Eisenia fetida was exposed to Cd and CIP alone or in combination at environmentally relevant concentrations. Cd and CIP accumulation in earthworm increased as their respective spiked concentrations increased. In fact, Cd accumulation increased by 39.7% when 1 mg/kg CIP was added; however, the addition of Cd did not affect CIP uptake. Compared with exposure to Cd alone, a greater ingestion of Cd following combined exposure to Cd and 1 mg/kg CIP resulted in greater oxidative stress and energy metabolism disturbances in earthworms. The reactive oxygen species (ROS) contents and apoptosis rate of coelomocytes were more sensitive to Cd than these biochemical indicators. In fact, 1 mg/kg Cd induced the derivation of ROS. Similarly, the toxicity of Cd (5 mg/kg) to coelomocytes was promoted by CIP (1 mg/kg), ROS content in coelomocytes and the apoptosis rate increased by 29.2% and 113.1%, respectively, due to increased Cd accumulation. Further investigation of the gut microorganisms revealed that the decreased abundance of Streptomyces strains (known as Cd accumulation taxa) could be a critical factor for enhanced Cd accumulation and greater Cd toxicity to earthworms following exposure to both Cd and CIP; this was because this microorganism group was eliminated by the simultaneous ingestion of CIP. This study stressed the role of gut microorganisms in altering the toxicity of Cd and CIP combined contamination in soil organisms. More attention should be paid to the ecological risks of such combined contamination in soils. [Display omitted] • Cd disturbed oxidative redox, energy metabolism, coelomocyte state of earthworm. • CIP enhance Cd toxicity to earthworm by promoting Cd uptake. • CIP eliminates Cd-resistant Streptomyces strains in gut to lead more Cd uptake. • Response of coelomocyte are sensitive to Cd stress than biochemical indicators. [ABSTRACT FROM AUTHOR]

Molybdenum disulfide (MoS 2 ) nanosheets are increasingly applied in several fields, but effective and accurate strategies to fully characterize potential risks to soil ecosystems are lacking. We introduce a coelomocyte-based in vivo exposure strategy to identify novel adverse outcome pathways (AOPs) and molecular endpoints from nontransformed (NTMoS 2 ) and ultraviolet-transformed (UTMoS 2 ) MoS 2 nanosheets (10 and 100 mg Mo/L) on the earthworm Eisenia fetida using nontargeted lipidomics integrated with transcriptomics. Machine learning-based digital pathology analysis coupled with phenotypic monitoring was further used to establish the correlation between lipid profiling and whole organism effects. As an ionic control, Na 2 MoO 4 exposure significantly reduced (61.2-79.5%) the cellular contents of membrane-associated lipids (glycerophospholipids) in earthworm coelomocytes. Downregulation of the unsaturated fatty acid synthesis pathway and leakage of lactate dehydrogenase (LDH) verified the Na 2 MoO 4 -induced membrane stress. Compared to conventional molybdate, NTMoS 2 inhibited genes related to transmembrane transport and caused the differential upregulation of phospholipid content. Unlike NTMoS 2 , UTMoS 2 specifically upregulated the glyceride metabolism (10.3-179%) and lipid peroxidation degree (50.4-69.4%). Consequently, lipolytic pathways were activated to compensate for the potential energy deprivation. With pathology image quantification, we report that UTMoS 2 caused more severe epithelial damage and intestinal steatosis than NTMoS 2 , which is attributed to the edge effect and higher Mo release upon UV irradiation. Our results reveal differential AOPs involving soil sentinel organisms exposed to different Mo forms, demonstrating the potential of liposome analysis to identify novel AOPs and furthermore accurate soil risk assessment strategies for emerging contaminants.

C-type lectins (CTLs) are Ca 2+ dependent carbohydrate-binding proteins that share structural homology in their carbohydrate-recognition domains (CRDs). In the present study, a novel CTL was identified from sea cucumber Apostichopus japonicus (named as Aj CTL-2). The deduced amino acid sequence of Aj CTL-2 was homologous to CTLs from other animals with the identities ranging from 33% to 40%. It contained a canonical signal peptide at the N-terminus, a low density lipoprotein receptor class A (LDLa), a C1r/C1s/Uegf/bone morphogenetic protein 1 (CUB), and a CRD with two motifs Glu-Pro-Asn (EPN) and Trp-Asn-Asp (WND) in Ca 2+ binding site 2. The mRNA transcripts of Aj CTL-2 were extensively expressed in all the tested tissues including respiratory tree, muscle, gut, coelomocyte, tube-foot, body wall and gonad, and the highest expression level of Aj CTL-2 in coelomocyte was about 4.2-fold ( p  < 0.05) of that in body wall. The mRNA expression level of Aj CTL-2 in coelomocyte increased significantly after Vibrio splendidus stimulation, and dramatically peaked at 12 h, which was 206.4-fold ( p  < 0.05) of that in control group. Aj CTL-2 protein was mainly detected in cytoplasm of coelomocyte by immunofluorescence. The recombinant Aj CTL-2 (r Aj CTL-2) displayed binding activity to d -galactose independent of Ca 2+ , while the binding activity to other tested pathogen-associated molecular patterns (PAMPs) including lipopolysaccharide (LPS), peptidoglycan (PGN), and mannose (Man) could not be detected. Surface plasmon resonance (SPR) analysis further revealed the high binding specificity and moderate binding affinity of r Aj CTL-2 to d -galactose (KD = 4.093 × 10 −6  M). After r Aj CTL-2 was blocked by its polyclonal antibody, the binding activity to d -galactose could not be detected by using a blocking ELISA (B-ELISA). Moreover, r Aj CTL-2 could bind various microorganisms including V. splendidus , V. anguillarum , Staphylococcus aureus , Bifidobacterium breve and Yarrowia lipolytica with the strongest binding activity to B. breve . These results collectively suggested that Aj CTL-2 was a member of CTL superfamily (CTLs) with preferential binding of d -galactose and participated in the immune response of sea cucumber. [ABSTRACT FROM AUTHOR]

Sea cucumber, Apostichopus japonicus , is one of the most important holothurian species cultured in China. Severe evisceration induced by various natural and artificial factors commonly occurs during transport and culture of A. japonicus. Evisceration causes higher mortality and lower yield. Along with the visceral regeneration process, sea cucumbers also regenerate coelomocytes in order to recover immune function. In this study, evisceration of A. japonicus was induced by intracoelomic injection of 0.35 M KCl. Regeneration of coelomocytes was investigated by time course cell counting as well as detection of DNA replication by the EdU labeling technique. Coelomic fluid volume was restored to the pre-evisceration level within 2 h after evisceration. Total coelomocyte count (TCC) reached a peak at 6 h post-evisceration, followed decreased and then increased with a slight fluctuation, restored to the pre-evisceration level at 35 d post-evisceration. The change in different subtypes of coelomocytes was consistent with that of total coelomocytes. However, there were some variations in the regeneration of coelomocyte subtypes. At the end of the study, only the counts of amoebocytes and morula cells recovered to the pre-evisceration level. DNA replication assay showed EdU-positive cells accounted for 9.5% before evisceration and 4.7% at 6 h post-evisceration. However, the percentage of EdU-positive cells significantly increased, reaching 18.6% at 3 d after evisceration, then declined. Therefore, we analyzed the observed increase in coelomocytes at 6 h post-evisceration, which may be due to coelomocyte migration from the water-vascular system into the coelom rather than de novo cell proliferation. [ABSTRACT FROM AUTHOR]

Sea urchin is one of marine animals with high economic and great scientific research values. Axial organ is a glandular organ that has been presumed as coelomocytes origin site. In this paper, two monoclonal antibodies (3G10 and 6B3) against coelomocytes of sea urchin Strongylocentrotus intermedius were developed by hybridoma technique. The mAbs were characterized by indirect immunofluorescence assay test (IIFAT), flow cytometry (FCM) and western blot assay. Results showed that mAb 3G10 recognized a protein of a molecular weight of 17 kDa in the spherule cells, while mAb 6B3 reacted with a protein of a molecular weight of 35 kDa in the phagocytes. Furthermore, specificity analysis revealed that the two mAbs could react with the coelomocytes of sea urchin S. nudus and Hemicentrotus pulcherrimus , but not with those of other common echinoderms including sea cucumber Apostichopus japonicus and starfish Asterias rollestoni. To determine whether the coelomocytes exist in the axial organ of sea urchin, the IIFAT assays were carried out based on the two mAbs. Result showed that positive fluorescence signals were distributed in the organ. It was revealed that the axial organ was rich in coelomocytes, which suggests that the organ may play as a producing source or reservoir in the ontogenesis of coelomocytes of sea urchin. [ABSTRACT FROM AUTHOR]

Proper development of a multicellular organism relies on well-coordinated regulation of cell fate specification, cell proliferation and cell differentiation. The C. elegans postembryonic mesoderm provides a useful system for uncovering factors involved in these processes and for further dissecting their regulatory relationships. The single Spalt-like zinc finger containing protein SEM-4/SALL is known to be involved in specifying the proliferative sex myoblast (SM) fate. We have found that SEM-4/SALL is sufficient to promote the SM fate and that it does so in a cell autonomous manner. We further showed that SEM-4/SALL acts through the SoxC transcription factor SEM-2 to promote the SM fate. SEM-2 is known to promote the SM fate by inhibiting the expression of two BWM-specifying transcription factors. In light of recent findings in mammals showing that Sall4, one of the mammalian homologs of SEM-4, contributes to pluripotency regulation by inhibiting differentiation, our work suggests that the function of SEM-4/SALL proteins in regulating pluripotency versus differentiation appears to be evolutionarily conserved. [ABSTRACT FROM AUTHOR]

Lysosomes, the major membrane-bound degradative organelles, have a multitude of functions in eukaryotic cells. Lysosomes are the terminal compartments in the endocytic pathway, though they display highly dynamic behaviors, fusing with each other and with late endosomes in the endocytic pathway, and with the plasma membrane during regulated exocytosis and for wound repair. After fusing with late endosomes, lysosomes are reformed from the resulting hybrid organelles through a process that involves budding of a nascent lysosome, extension of the nascent lysosome from the hybrid organelle, while remaining connected by a membrane bridge, and scission of the membrane bridge to release the newly formed lysosome. The newly formed lysosomes undergo cycles of homotypic fusion and fission reactions to form mature lysosomes. In this study, we used a forward genetic screen in Caenorhabditis elegans to identify six regulators of lysosome biology. We show that these proteins function in different steps of lysosome biology, regulating lysosome formation, lysosome fusion, and lysosome degradation. [ABSTRACT FROM AUTHOR]

Echinoderms have many types of coelomocytes, which have been known to form aggregates immediately after they are removed from the coelom. To assess the roles that each type of coelomocyte plays in aggregate formation, cellular components of coelomocyte aggregates of the Japanese sea cucumber, Apostichopus japonicus , were investigated. The coelomocytes were tentatively classified into 12 types based on May-Grunwald/Giemsa staining. After the coelom was incubated for 30 min or 6 h, the aggregates were disaggregated completely with 200 mM EDTA. Differential counts of the dissociated cells indicated that the largest component of the aggregates was amoebocytes (67.8%) and the second-largest component of the aggregates incubated 30 min was a type of basophilic granulocyte. In the 6h-incubated aggregates, the fraction of amoebocytes decreased to 59.0%, while that of lymphoid cells significantly increased, which suggests that lymphoid cells participate in late-stage aggregation. After 24-h incubation, only a portion of the aggregated cells could be disaggregated with EDTA. After 48 h, most of the cells could not be detached from the aggregates. Microscopy of frozen sections of the aggregates after 6-h incubation revealed that amoebocytes constructed a mesh-like structure to which other types of cells adhered. After 48 h, the borders of the cells and the intracellular granules were not recognizable. In time-lapse microscopy, the aggregates were observed to move on a glass slide, which suggests that aggregates can “crawl” on the intraluminal surface of the coelom toward, for example, injured regions in the body of the sea cucumber. [ABSTRACT FROM AUTHOR]

A comparative analysis of the morphology of in vitro flattened coelomocytes of two starfish species, Aphelasterias japonica and Patiria pectinifera (Echinodermata: Asteroidea), and hemocytes of the bivalve Callista brevisiphonata (Mollusca: Bivalvia) was performed using a number of nonlinear parameters including several types of fractal dimensions and lacunarities. The visually 'chaotic' shapes of in vitro flattened hemocytes and coelomocytes of the studied marine invertebrate species significantly differ in a number of nonlinear parameters. This fact allows numerical description of the morphology of hemolymph cells of the studied animals and gives grounds to assume a species specificity of the biological differences that influence the morphology of in vitro flattened cells. [ABSTRACT FROM AUTHOR]

The tube-within-tube body plan of earthworms is appropriate for studying the interactions of microorganisms with the immune system of body cavities such as the digestive tract and coelom. This study aims to describe the immune response on the molecular and cellular level in the coelomic cavity and the gut of the earthworm Eisenia andrei after experimental microbial challenge by administering two bacterial strains ( Escherichia coli and Bacillus subtilis ) or yeast Saccharomyces cerevisiae to the environment. The changes in mRNA levels of defense molecules (pattern recognition receptor CCF, lysozyme, fetidin/lysenins) in the coelomocytes and gut tissue were determined by quantitative PCR. The immune response at a cellular level was captured in histological sections, and the expression of CCF was localized using in situ hybridization. Coelomocytes respond to the presence of bacteria in the coelomic cavity by increasing the mRNA levels of defense molecules, especially CCF. The immune response in gut tissue is less affected by microbial stimulation because the epithelial cells of gut exhibit basically strong mRNA synthesis of ccf as a defense against the continuous microbial load in the gut lumen. The cellular immune response is mediated by coelomocytes released from the mesenchymal lining of the coelomic cavity. These combined immune mechanisms are necessary for the survival of earthworms in the microbially rich environment of soil. [ABSTRACT FROM AUTHOR]

Metchnikoff’s use of sea star larvae to observe encapsulation and phagocytosis, which was followedmuch later by allograft rejection kinetics, revealed that echinoderms had an innate immune system thatwas lacking of adaptive attributes. Larval sea urchins mount defenses in response to contact withmicrobes, which are mediated by phagocytic blastocoelar cells and pigment cells. In the adult, thecoelomocytes mediate immune responses through phagocytosis and encapsulation of foreign particles inaddition to degranulation of antimicrobial molecules. Molecular analysis of immune functions in the seaurchin has demonstrated a complement system that appears to have multiple alternative pathways andseveral activators of the lectin pathway, but may be missing the terminal pathway. Other genes andproteins involved in the sea urchin immunity include expanded sets of lectins, proteins with scavengerreceptor cysteine-rich repeats, Toll-like receptors and associated signalling proteins. A vast array ofproteins belonging to the 185/333 family are expressed in coelomocytes in response to lipopolysaccharideand show a surprising level of diversity. The sea urchin innate immune system has a number of largegene families with unexpected complexities and elevated levels of diversification.

Three proteins with PO-like activities in the coelomocytes of sea urchin Strongylocentrotus nudus were identified using electrophoretic method and named as SnPO1, SnPO2 and SnPO3 according to their molecular mass from high to low. The SnPOs were characterized for substrate specificity and the effects of temperature, pH, divalent metal ions and inhibitors on PO activities. They showed oxidative activities to L-3,4-dihydroxyphenylalanine. ( l -DOPA), dopamine and hydroquinone, but failed to oxidize tyrosine, which illustrated the three proteins had laccase-like PO activities. The optimum temperature for the activities of SnPO1, SnPO2 and SnPO3 was 75 °C, 70 °C, 40 °C, and the optimum pH was 7.0, 9.0, 8.0, respectively. The SnPOs were notably activated after being incubated in boiled water for 60 min, suggesting that the three proteins are thermophilic. The activity of SnPO1 was greatly enhanced by Cu 2+ , Mn 2+ and Fe 2+ and inhibited by Pb 2+ , Cd 2+ , EDTA, DETC, sodium sulfite and ascorbic acid, but SnPO2 and SnPO3 were not obviously affected by Pb 2+ and Cd 2+ , suggesting the three proteins are copper-containing, and the catalytic properties of SnPO1 might be different from those of SnPO2 and SnPO3. Taken together, SnPO1, SnPO2 and SnPO3 might play different roles in the immune and physiological processes of S. nudus . [ABSTRACT FROM AUTHOR]

Despite the lack of knowledge of their exact effects, peloids (natural muds) are widely applied in clinical treatment and prevention of different diseases, especially in rheumatic and gynecological disorders or skin diseases. Primarily we have information on their inorganic components, but only limited data are available on the organic components and nothing on their mechanism of chemical action. The objective of the present study was to detect the DNA-damaging effects (possible genotoxic effect) of peloid samples using the single-cell comet assay on Long Evans rat lymphocytes, human lymphocytes, and Eisenia fetida coelomocytes. Rat and human lymphocytes were exposed to the in toto peloid samples, in vitro. The Eisenia cells were extracted from the coelom of animals kept in the intact peloid sample. An indicator derived from the DNA fluorescence intensity was used in the statistical evaluation. The predominantly organic (Hévíz) sample showed a significant alteration from the negative control in several cases, while the inorganic (Kolop) applied did not. A higher quantity of organic compounds may have an important role in the emergence of DNA damage. The results revealed that medical muds have not only positive health effects but can also contain substances with potential human toxicity risk. Our research provides essential steps towards the creation of a toxicity profile and the possible safe use of peloids as medicinal therapy. [ABSTRACT FROM AUTHOR]

The essential mechanism within immune systems is the recognition of pathogens and parasites by the immune system cells, which attach to their targets and destroy them. Glycans are fundamental macromolecular components of all cells, and are important in the vertebrate immunity. But, glycans have been investigated rarely in coelomocytes of echinoids. Hence, the aim of this study is to determine the monosaccharides which form glycan chains on the sea urchin immune system cells, coelomocytes, via analytical and lectin histochemistry methods. The study material is the coelomocytes obtained from adult sea urchin Paracentrotus lividus . In order to analyze the monosaccharides with the Capillary Liquid Chromatography Electrospray Ionization Tandem Mass Spectrometry (CapLC-ESI-MS/MS) system, the samples underwent hydrolysation, reacetylation and derivatization steps. In order to determine the monosaccharides with the lectin histochemistry, the cells were incubated with fluorescein isothiocyanate (FITC) conjugated PNA, HPA, WGA-suc, WGA, and PSL lectins and then photographed with the fluorescence microscope. As a result of the CapLC-ESI-MS/MS analysis; mannose, ribose, N-acetylglucosamine, glucose, N-acetylgalactosamine, galactose, arabinose, xylose and fucose monosaccharides were detected. A peak area calculation analysis revealed the most prevalent saccharides as glucose, galactose and fucose, respectively. Lectin histochemistry came out with higher intensity emission signals obtained from the FITC-conjugated lectin WGA, which is specific to N-acetylglucosamine and sialic acid in comparison to the emission obtained from the sialic acid unspecific WGA-suc lectin. This finding indicates the existence of sialic acid within coelomocytes. Fluorescent emissions from other lectins were detected at lower levels. Determination of the monosaccharides which form glycan chains of the sea urchin coelomocytes and elucidating their similarities among other invertebrate and vertebrate systems is vital in terms of understanding the uncovered complex features of the immune systems of higher vertebrates. [ABSTRACT FROM AUTHOR]

The polian vesicle and coelom of sea cucumber Apostichopus japonicus were full of coelomic fluid in which many types of coelomocytes with different functions were suspended. Our previous work has indicated the differences of coelomocytes between two sites mainly in subtype proportion, non-specific immune enzymes activities and several immune-related genes expression levels in healthy A. japonicus . However, the functional similarities and differences of coelomic fluid in two sites including the coelom and polian vesicle after pathogenic infection still remain unclear. Here, we investigated the changes of the total coelomocyte density (TCD) and differential coelomocyte density (DCD) after pathogen infection by Vibrio splendidus in coelom and polian vesicle. After infected by V. splendidus , the TCD in the coelom and polian vesicle rapidly declined at 12 h, and then the TCD in the coelom showed a stably ascending trend, while the TCD in the polian vesicle reached a peak at 24 h post infection (hpi), and then showed a continuously decline trend from 24 hpi to 72 hpi followed by a slow elevation until recovering the normal level from 72 hpi to 96 hpi. Then the activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD) were determined to evaluate the response of cell-free coelomic fluid to V. splendidus infection. The activities of ACP, AKP and CAT showed similar trends in the coelom and polian vesicle. The SOD activity significantly increased in the polian vesicle, whereas it exhibited a decreasing trend in the coelom. Finally, the expression profiles of nine immune-related genes including Aj-MyD88, Aj-IRAK4, Aj-i-Lys, Aj-Rel, Aj-p50, Aj-DMBT1, Aj-CDC, Aj-Rrp15 and Aj-Fibrinogen C were detected after V. splendidus challenge. The results suggested all the detected genes were significantly up-regulated both in the coelom and polian vesicle, and the expression levels of these genes in two sites shared similar trends except Aj-MyD88 and Aj-DMBT1 . This research provides a new insight into the differentially immune roles of coelomic fluid and coelomocytes in polian vesicle and coelom response to bacterial infections and supplements comprehensive resources for better understanding the innate immune response of A. japonicus ., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 The Author(s).)

During rearing in hatcheries and transportation to restocking sites, sea cucumbers are often exposed to air for several hours, which may depress their non-specific immunity and lead to mass mortality. We performed transcriptome analysis of Apostichopus japonicus coelomocytes after air exposure to identify stress-related genes and pathways. After exposure to air for 1 h, individuals were re-submerged in aerated seawater and coelomocytes were collected at 0, 1, 4, and 16 h (B, H1, H4, and H16, respectively). We identified 6148 differentially expressed genes, of which 3216 were upregulated and 2932 were downregulated. Many genes involved in the immune response, antioxidant defense, and apoptosis were highly induced in response to air exposure. Enrichment analysis of Gene Ontology terms showed that the most abundant terms in the biological process category were oxidation-reduction process, protein folding and phosphorylation, and receptor-mediated endocytosis for the comparison of H1 vs. B, H4 vs. H1, and H16 vs. H4, respectively. Kyoto Eecyclopedia of Genes and Genomes enrichment analysis showed that six pathways related to the metabolism of proteins, fats, and carbohydrates were shared among the three comparisons. These results indicated that sea cucumbers regulate the expression of genes related to the antioxidant system and energy metabolism to resist the negative effects of air exposure stress. These findings may be applied to optimize juvenile sea cucumber production, and facilitate molecular marker-assisted selective breeding of an anoxia-resistant strain. • In air exposure stress, sea cucumber regulates anti-oxidation system to maintain basic life. • In sea cucumber, immune related genes were activated to repair air exposure stress injury. • The expression of hif - 1α increased in A. japonicus coelomocytes under air exposure stress. • hsp70 expression increased to protect the sea cucumber from air exposure stress. • Apoptosis process may be a important mechanism to protect sea cucumber from air exposure stress. [ABSTRACT FROM AUTHOR]