RESUMO
Due to their tissue structure similar to mammalian skin and their close evolutionary relationship with chordates, holothurians (Echinodermata: Holothuroidea) are particularly interesting for studies on wound healing. However, previous studies dealing with holothuroid wound healing have had limited approaches, being restricted to tissue repair or perivisceral immune response. In this study, we combined tissue, cellular and humoral parameters to study the wound healing process of Holothuria grisea. The immune responses of the perivisceral coelom were assessed by analyzing the number, proportion and viability of coelomocytes and the volume and protein concentration of the coelomic fluid. Additionally, the morphology of the healing tissue and number of coelomocytes in the connective tissue of different body wall layers were examined over 30 days. Our results showed that perivisceral reactions started 3 h after injury and decreased to baseline levels within 24 h. In contrast, tissue responses were delayed, beginning after 12 h and returning to baseline levels only after day 10. The number of coelomocytes in the connective tissue suggests a potential cooperation between these cells during wound healing: phagocytes and acidophilic spherulocytes act together in tissue clearance/homeostasis, whereas fibroblast-like and morula cells cooperate in tissue remodeling. Finally, our results indicate that the major phases observed in mammalian wound healing are also observed in H. grisea, despite occurring at a different timing, which might provide insights for future studies. Based on these data, we propose a model that explains the entire healing process in H. grisea.
Assuntos
Holothuria , Cicatrização , Animais , Cicatrização/fisiologia , Holothuria/fisiologia , Imunidade HumoralRESUMO
BACKGROUND: Molecules and cytokines can be targeted in cancer therapy. Transforming growth factor-beta (TGF-ß) is a cytokine that acts on protein kinase receptors in the plasma membrane. The signaling pathway of TGF-ß can trigger the phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) pathway, a signal transduction pathway important in cancer growth and development. However, this PI3K/AKT cascade can be inhibited by phosphatase and tensin homolog (PTEN) tumor suppressor genes. AIM: To determine the inhibitory effect of Holothuria scabra methanol extract (HSE) on breast cancer growth through the TGF-ß/PI3K pathways and PTEN tumor suppressor gene on a breast cancer (BC) mice model. MATERIALS AND METHODS: Female C57BL6 mice were subcutaneously injected with carcinogen DMBA 1 mg/kg body weight (BW) and fed a high-fat diet (HFD). Mice were randomly divided into five groups (n = 6): negative control (NC) administered with a standard diet, positive control (PC) administered with DMBA and HFD, and three treatment groups (T1, T2, and T3) treated with HSE doses of 0.33, 0.66, and 0.99 g/kg BW for 12 weeks. TGF-ß concentration in the blood serum of mice was assessed by ELISA and the PIK3CA and PTEN gene expression by qRT-PCR. RESULTS: The treatment with HSE resulted in a significant decrease in TGF-ß concentrations in the blood sera of treatment groups T1 (35.31 ± 17.33), T2 (43.31 ± 17.42), and T3 (48.67 ± 20.94) pg/mL compared to the PC group (162.09 ± 11.60) pg/mL (p < 0.001). However, only HSE at a dose of 0.99 g/kg BW decreased the PIK3CA gene expression (p = 0.026), and at a dose of 0.66 g/kg BW increased the PTEN expression up to 4.93-fold. CONCLUSION: HSE is capable of inhibiting the TGF-ß/PIK3CA pathway and increasing the PTEN gene expression.
Assuntos
PTEN Fosfo-Hidrolase , Fosfatidilinositol 3-Quinases , Extratos Vegetais , Transdução de Sinais , Fator de Crescimento Transformador beta , Animais , PTEN Fosfo-Hidrolase/metabolismo , Feminino , Fator de Crescimento Transformador beta/metabolismo , Camundongos , Transdução de Sinais/efeitos dos fármacos , Extratos Vegetais/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Holothuria/química , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Metanol/química , Modelos Animais de Doenças , Camundongos Endogâmicos C57BL , Humanos , Proliferação de Células/efeitos dos fármacosRESUMO
Holothuria scabra, a commercially valuable yet ecologically vulnerable tropical holothuroid, has experienced a severe decline in its wild populations, especially in China. Genomic resources are crucial for the development of effective genomic breeding projects and stock conservation strategies to restore these natural populations. Until now, a high-quality, chromosome-level reference genome for H. scabra has not been available. Here, we employed Oxford Nanopore and Hi-C sequencing technologies to assemble and annotate a high-quality, chromosome-level reference genome of H. scabra. The final genome comprised 31 scaffolds with a total length of 1.19 Gb and a scaffold N50 length of 53.52 Mb. Remarkably, 1,191.67 Mb (99.95%) of the sequences were anchored to 23 pseudo-chromosomes, with the longest one spanning 79.75 Mb. A total of 34,418 protein-coding genes were annotated in the final genome, with BUSCO analysis revealing 98.01% coverage of metazoa_odb10 genes, marking a significant improvement compared to the previous report. These chromosome-level sequences and annotations will provide an essential genomic basis for further investigation into molecular breeding and conservation management of H. scabra.
Assuntos
Cromossomos , Genoma , Holothuria , Anotação de Sequência Molecular , Animais , Holothuria/genética , ChinaRESUMO
Aggregative α-synuclein and incurring oxidative stress are pivotal cascading events, leading to dopaminergic (DAergic) neuronal loss and contributing to clinical manifestations of Parkinson's disease (PD). Our previous study demonstrated that 2-butoxytetrahydrofuran (2-BTHF), isolated from Holothuria scabra (H. scabra), could inhibit amyloid-ß aggregation and its ensuing toxicity, which leads to Alzheimer's disease. In the present study, we found that 2-BTHF also attenuated the aggregative and oxidative activities of α-synuclein and lessened its toxicity in a transgenic Caenorhabditis elegans (C. elegans) PD model. Such worms treated with 100 µM of 2-BTHF showed substantial reductions in α-synuclein accumulation and DAergic neurodegeneration. Mechanistically, 2-BTHF, at this concentration, significantly decreased aggregation of monomeric α-synuclein and restored locomotion and dopamine-dependent behaviors. Molecular docking exhibited potential bindings of 2-BTHF to HSF-1 and DAF-16 transcription factors. Additionally, 2-BTHF significantly increased the mRNA transcripts of genes encoding proteins involved in proteostasis, including the molecular chaperones hsp-16.2 and hsp-16.49, the ubiquitination/SUMOylation-related ubc-9 gene, and the autophagy-related genes atg-7 and lgg-1. Transcriptomic profiling revealed an additional mechanism of 2-BTHF in α-synuclein-expressing worms, which showed upregulation of PPAR signaling cascades that mediated fatty acid metabolism. 2-BTHF significantly restored lipid deposition, upregulated the fat-7 gene, and enhanced gcs-1-mediated glutathione synthesis in the C. elegans PD model. Taken together, this study demonstrated that 2-BTHF could abrogate aggregative and oxidative properties of α-synuclein and attenuate its toxicity, thus providing a possible therapeutic application for the treatment of α-synuclein-induced PD.
Assuntos
Caenorhabditis elegans , Furanos , Holothuria , Estresse Oxidativo , alfa-Sinucleína , Animais , Humanos , alfa-Sinucleína/metabolismo , Animais Geneticamente Modificados , Caenorhabditis elegans/efeitos dos fármacos , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Modelos Animais de Doenças , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/metabolismo , Furanos/farmacologia , Simulação de Acoplamento Molecular , Estresse Oxidativo/efeitos dos fármacos , Doença de Parkinson/metabolismo , Doença de Parkinson/tratamento farmacológicoRESUMO
With the continuous rise of the sea cucumber aquaculture industry in China, the tropical sea cucumber aquaculture industry is also improving. However, research on the gut microorganisms of tropical sea cucumbers in captivity is scarce. In this study, high-throughput sequencing methods were used to analyze the gut microbial composition of Stichopus monotuberculatus and Holothuria scabra in the dry season and wet season of artificial environments. The results showed that 66 phyla were obtained in all samples, of which 59 phyla were obtained in the dry season, and 45 phyla were obtained in the wet season. The Tax4Fun analysis showed that certain gut bacterial communities affect the daily metabolism of two sea cucumber species and are involved in maintaining gut microecological balance in the gut of two sea cucumber species. In addition, compared with differences between species, PCoA and UPGMA clustering analysis showed the gut prokaryotes of the same sea cucumber species varied more in different seasons, indicating that the influence of environment was higher than the feeding choices of sea cucumbers under relatively closed conditions. These results revealed the gut bacterial community composition of S. monotuberculatus and H. scabra and the differences in gut bacterial structure between two sea cucumber species in different seasons were compared, which would provide the foundation for tropical sea cucumber aquaculture in the future.
Assuntos
Bactérias , Microbioma Gastrointestinal , Pepinos-do-Mar , Estações do Ano , Animais , Microbioma Gastrointestinal/genética , Bactérias/classificação , Bactérias/genética , Pepinos-do-Mar/microbiologia , Pepinos-do-Mar/genética , Aquicultura , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Holothuria/microbiologia , Holothuria/genética , Stichopus/microbiologia , Stichopus/genética , RNA Ribossômico 16S/genéticaRESUMO
BACKGROUND: The white teatfish, Holothuria fuscogilva, is widely distributed in coastal areas, including waters around coral reefs and seagrasses in the Indo-Pacific. In Kenya, the species is distributed in shallow reefs with higher landings reported from the Vanga-Shimoni-Gazi seascape on the Kenyan south coast. Despite its high exploitation for export and its vulnerable and endangered statuses under IUCN and CITES respectively, Kenya's H. fuscogilva populations and how they may have been impacted by the fishing pressure have not been studied. METHODS: We estimated the genetic diversity and structure of H. fuscogilva population conveniently sampled from three sites in Kenyan south coast using the mitochondrial cytochrome oxidase subunit I (COI) gene sequences. We recorded 30 haplotypes with 43 polymorphic sites across the population. Furthermore, we estimated an overall high haplotype diversity and low nucleotide diversity of estimates of h = 0.970 ± 0.013 and π = 0.010 ± 0.001 respectively. CONCLUSIONS: These preliminary findings suggest several population outcomes, among them a fit population, which require confirming with more comprehensive study to inform strategies for the sustainable exploitation and management of the species.
Assuntos
Holothuria , Animais , Holothuria/genética , Quênia , Variação Genética/genética , Genética Populacional , Genes Mitocondriais , Haplótipos/genética , DNA Mitocondrial/genéticaRESUMO
With global climate changing, hypo-salinity events are increasing in frequency and duration because of continuous rainfall and freshwater inflow, which causes reduced cytosolic osmolarity and cellular stress responses in aquatic animals. Sea cucumbers are considered stenohaline because they lack osmoregulatory organs and are vulnerable to salinity fluctuations. In this study, we performed multiple biochemical assays, de novo transcriptomics, and widely targeted metabolomics to comprehensively explore the osmoregulatory mechanisms and physiological responses of sea cucumber Holothuria moebii to hypo-osmotic stress, which is a representative specie that is frequently exposed to hypo-saline intertidal zones. Our results found that H. moebii contracted their ambulacral feet and oral tentacles, and the coelomic fluid ion concentrations were reduced to be consistent with the environment. The microvilli of intestines and respiratory trees underwent degeneration, and the cytoplasm exhibited swelling and vacuolation. Moreover, the Na+, K+, and Cl- concentrations and Na+/K+-ATPase activity were significantly reduced under hypo-osmotic stress. The decrease in protein kinase A activity and increase in 5'-AMP level indicated a significant inhibition of the cAMP signaling pathway to regulate ion concentrations. And small intracellular organic molecules (amino acids, nucleotides and their derivatives) also play crucial roles in osmoregulation through oxidative deamination of glutamate, nucleotide catabolism, and nucleic acid synthesis. Moreover, lysosomes and peroxisomes removed oxidative damage, whereas antioxidant metabolites, such as N-acetyl amino acids and glutathione, were increased to resist oxidative stress. With prolonged hypo-osmotic stress, glycerophospholipid metabolism was enhanced to maintain membrane stability. Furthermore, acyl-CoA-binding protein activity was significantly inhibited, and only a small amount of acylcarnitine was significantly accumulated, which indicated a disruption in energy metabolism. PPAR signaling pathway and choline content were up-regulated to promote fatty acid metabolism under hypo-osmotic stress. Overall, our results provide new insights into the osmoregulatory mechanisms and physiological responses of sea cucumbers to hypo-osmotic stress.
Assuntos
Antioxidantes , Metabolismo Energético , Holothuria , Osmorregulação , Pressão Osmótica , Animais , Holothuria/fisiologia , Antioxidantes/metabolismo , SalinidadeRESUMO
Abstract Introduction: The sea cucumber Holothuria (Halodeima) grisea Selenka, 1867 is a common echinoderm in intertidal regions along the Brazilian coast, which recently became the focus of unreported and unregulated fisheries. This study was carried out in sandy-rocky substrates at Armação do Itapocoroy, Penha, Santa Catarina (26o47' S; 48o36' W), near its southern limit of geographic distribution. Objective: To determine the densities (individuals*m-2) of Holothuria (H.) grisea within a spatial-temporal perspective as well as to determine biometric and growth characteristics of the population. Methods: Two-meter wide transects perpendicular to the coastline were carried out in winter and spring 2019 and in summer and spring 2020, in periods of spring low-tides. In each sampling occasion the total number of specimens of H. grisea were determined, and a group of 90 organisms was submitted to in situ biometrics (weight, length and width), and immediately returned alive to their habitat. Results: The densities of H. (H.) grisea were significantly higher in the subtidal sector and lower in the upper intertidal sector with no indication of significant differences among sampling campaigns. Depth was the primary factor explaining the observed density patterns and rugosity of the substrate was secondary but also important. The body length ranged from 5.2 to 22.5 cm, whereas the weight varied from 6.0 to 230 g. The mean and modal lengths were 12.54 and 13 cm, respectively. Approximately 75 % of the population sampled was between 10 and 14 cm and the average weight was 60 g. Estimates from von Bertalanffy growth function indicate that the youngest sea cucumber was one year-old, and the oldest had approximately two and a half years. Conclusions: This is the first study to determine biometric parameters for H. (H.) grisea in southern Brazil and the first one to estimate growth and age estimates for a wild population of this species. The densities recorded in the present study were lower than those previously reported for this region, suggesting anthropic influence.
Resumen Introducción: El pepino de mar Holothuria (Halodeima) grisea Selenka, 1867 es un equinodermo común en las regiones intermareales a lo largo de la costa brasileña, que recientemente se convirtió en foco de pesquerías no declaradas y no reguladas. Este estudio se realizó en sustratos arenosos-rocosos en Armação do Itapocoroy, Penha, Santa Catarina (26o47' S; 48o36' W), cerca del límite sur de su distribución geográfica. Objetivo: Determinar las densidades (individuos*m-2) de Holothuria (H.) grisea dentro de una perspectiva espacio-temporal así como determinar las características biométricas y de crecimiento de la población. Métodos: Se realizaron transectos de dos metros de ancho perpendiculares a la línea de costa en invierno y primavera de 2019 y en verano y primavera de 2020, en periodos de bajamar sicigia. En cada ocasión de muestreo se determinó el número total de especímenes de H. (H.) grisea, y se sometió un grupo de 90 organismos a biometría in situ (peso, longitud y ancho), e inmediatamente se los devolvieron vivos a su hábitat. Resultados: Las densidades de H. (H.) grisea fueron significativamente más altas en el sector submareal y más bajas en el sector intermareal superior sin indicios de diferencias significativas entre las campañas de muestreo. La profundidad fue el factor principal que explica los patrones de densidad observados y la rugosidad del sustrato fue secundaria pero también importante. La longitud del cuerpo varió de 5.2 a 22.5 cm, mientras que el peso varió de 6.0 a 230 g. Las longitudes media y modal fueron 12.54 y 13 cm, respectivamente. Aproximadamente el 75 % de la población muestreada midió entre 10 y 14 cm y el peso promedio fue de 60 g. Estimados de la función de crecimiento de von Bertalanffy indican que el ejemplar más joven presentaba un año de edad, mientras el más viejo presentaba cerca de dos años y medio. Conclusiones: Este es el primer estudio que determina parámetros biométricos para una población de H. (H.) grisea en el sur de Brasil y el primero en estimar el crecimiento y edades para una población salvaje de esta especie. Las densidades registradas en el presente estudio fueron inferiores a las reportadas previamente para esta región sugiriendo la ocurrencia de influencia antrópica.
Assuntos
Animais , Dinâmica Populacional , Holothuria/crescimento & desenvolvimento , Estudos de AmostragemRESUMO
Fungi associated with the marine echinoderm, Holothuria scabra, produces extracellular enzymes and bioactive metabolites, and mycoviruses that could be used for biotechnological and pharmaceutical applications. The species identification based on molecular and morphological characteristics classified the culturable fungi into twenty-three genera belonging to eight orders, Chaetothyriales, Eurotiales, Hypocreales, Mucorales, Mycosphaerellales, Onygenales, Pleosporales and Venturiales, from four classes, Eurotiomycetes, Dothideomycetes, Mucoromycetes and Sordariomycetes of the two phyla Ascomycota and Mucoromycota. The most frequent genera were Aspergillus (relative frequency, 45.30%) and Penicillium (relative frequency, 22.68%). The Menhinick species richness and Shannon species diversity indices were 1.64 and 2.36, respectively, indicating a high diversity of fungi. An enzymatic production test revealed that sixteen isolates could produce proteases and amylases at different levels. The presence of mycoviruses was detected in eight isolates with different genomic profiles. Thirty-two of the 55 isolates produced antimicrobial metabolites which had an inhibitory effect on various microbial pathogens. Most of these active isolates were identified as Aspergillus, Penicillium and Trichoderma. Notably, Aspergillus terreus F10M7, Trichoderma harzianum F31M4 and T. harzianum F31M5 showed the most potent activity against both Gram-positive and Gram-negative bacteria and human pathogenic fungi. Our study represents the first report of the mycobiota associated with the marine echinoderm Holothuria scabra.
Assuntos
Ascomicetos , Holothuria , Penicillium , Pepinos-do-Mar , Animais , Humanos , Holothuria/microbiologia , Antibacterianos/metabolismo , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Fungos , Ascomicetos/genética , Penicillium/genética , FilogeniaRESUMO
BACKGROUND: Sea cucumbers exhibit a remarkable ability to regenerate damaged or lost tissues and organs, making them an outstanding model system for investigating processes and mechanisms of regeneration. They can also reproduce asexually by transverse fission, whereby the anterior and posterior bodies can regenerate independently. Despite the recent focus on intestinal regeneration, the molecular mechanisms underlying body wall regeneration in sea cucumbers still remain unclear. RESULTS: In this study, transverse fission was induced in the tropical sea cucumber, Holothuria leucospilota, through constrainment using rubber bands. Histological examination revealed the degradation and loosening of collagen fibers on day-3, followed by increased density but disorganization of the connective tissue on day-7 of regeneration. An Illumina transcriptome analysis was performed on the H. leucospilota at 0-, 3- and 7-days after artificially induced fission. The differential expression genes were classified and enriched by GO terms and KEGG database, respectively. An upregulation of genes associated with extracellular matrix remodeling was observed, while a downregulation of pluripotency factors Myc, Klf2 and Oct1 was detected, although Sox2 showed an upregulation in expression. In addition, this study also identified progressively declining expression of transcription factors in the Wnt, Hippo, TGF-ß, and MAPK signaling pathways. Moreover, changes in genes related to development, stress response, apoptosis, and cytoskeleton formation were observed. The localization of the related genes was further confirmed through in situ hybridization. CONCLUSION: The early regeneration of H. leucospilota body wall is associated with the degradation and subsequent reconstruction of the extracellular matrix. Pluripotency factors participate in the regenerative process. Multiple transcription factors involved in regulating cell proliferation were found to be gradually downregulated, indicating reduced cell proliferation. Moreover, genes related to development, stress response, apoptosis, and cell cytoskeleton formation were also involved in this process. Overall, this study provides new insights into the mechanisms of whole-body regeneration and uncover potential cross-species regenerative-related genes.
Assuntos
Holothuria , Pepinos-do-Mar , Animais , Pepinos-do-Mar/genética , Holothuria/genética , Regeneração/genética , Perfilação da Expressão Gênica , Fatores de Transcrição/genéticaRESUMO
A sulfated polysaccharide (AG) was extracted and isolated from the sea cucumber H. fuscopunctata, consisting of GlcNAc, GalNAc, Gal, Fuc and lacking any uronic acid residues. Importantly, several chemical depolymerization methods were used to elucidate the structure of the AG through a bottom-up strategy. A highly sulfated galactose (oAG-1) and two disaccharides labeled with 2,5-anhydro-D-mannose (oAG-2, oAG-3) were obtained from the deaminative depolymerized product along with the structures of the disaccharide derivatives (oAG-4~oAG-6) identified from the free radical depolymerized product, suggesting that the repeating building blocks in a natural AG should comprise the disaccharide ß-D-GalS-1,4-D-GlcNAc6S. The possible disaccharide side chains (bAG-1) were obtained with mild acid hydrolysis. Thus, a natural AG may consist of a keratan sulfate-like (KS-like) glycosaminoglycan with diverse modifications, including the sulfation types of the Gal residue and the possible disaccharide branches α-D-GalNAc4S6S-1,2-α/ß-L-Fuc3S linked to the KS-like chain. Additionally, the anticoagulant activities of the AG and its depolymerized products (dAG1-9) were evaluated in vitro using normal human plasma. The AG could prolong activated partial thromboplastin time (APTT) in a dose-dependent manner, and the activity potency was positively related to the chain length. The AG and dAG1-dAG3 could prolong thrombin time (TT), while they had little effect on prothrombin time (PT). The results indicate that the AG could inhibit the intrinsic and common coagulation pathways.
Assuntos
Holothuria , Pepinos-do-Mar , Animais , Humanos , Sulfato de Queratano/química , Holothuria/química , Pepinos-do-Mar/química , Polissacarídeos/farmacologia , Polissacarídeos/química , Dissacarídeos , Anticoagulantes/químicaRESUMO
The overexploitation of the western Pacific Ocean has expanded the sea cucumber fishery into new regions to supply the Asian market. In 2013, sea cucumbers were removed from the Croatian marine protected species list, and commercial fishery took place for a short period (2017-2018) in the Eastern Adriatic Sea. However, holothuroid species are difficult to distinguish. Holothuria mammata is a species that has rarely been reported in this region and strongly resembles the common species Holothuria tubulosa. This is the first study to assess the genetic diversity of sea cucumbers in the Adriatic Sea using genetic barcoding of the mitochondrial gene cytochrome c oxidase subunit 1 (COI). Specimens for barcoding were collected from the northern and central Adriatic, along with a specimen that had been previously identified as H. sp. cf. mammata based on its morphological characteristics. While genetic analyses showed identified this specimen as H. tubulosa, 30% of the collected specimens were genetically identified as H. mammata. These results call into question the historically accepted sea cucumber assemblage in the Adriatic Sea, which regarded H. mammata as a rare species and generally disregarded its presence in large census studies. Such species distribution data are extremely important in developing and monitoring a sustainable fishery.
Assuntos
Holothuria , Pepinos-do-Mar , Animais , Pepinos-do-Mar/genética , Holothuria/genética , Oceano PacíficoRESUMO
The sea cucumber Holothuria leucospilota is an economically and ecologically important tropical species. Following development into juveniles, H. leucospilota undergoes a color change from white to black, involving a pigmentation process for over a period of several months. In this study, a combination of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and Next-Generation sequencing (NGS) were employed to investigate the changes in metabolomic and transcriptomic profiles during pigmentation in H. leucospilota juveniles. The metabolomic analysis identified a total of 341 metabolites, of which 52 were found to be differentially regulated (P < 0.05 and VIP > 1), with 27 being upregulated in white individuals and 25 in black individuals. Additionally, 632 differentially expressed genes (DEGs) were identified, with 380 genes upregulated in white samples and 252 genes upregulated in black samples. Interestingly, the melanin content and tyrosinase transcript levels did not display significant differences between the two groups. Metabolomic data suggested the involvement of the linoleic acid metabolic pathway in pigmentation. Transcriptomic analysis, coupled with realtime PCR validation, revealed a decrease in the transcript levels of digestive enzymes like α-amylase, maltase-glucoamylase, and trehalase after the juveniles changed to black. Furthermore, the mRNA expressions of major yolk proteins showed a decline, indicating a shift in the accumulation of protein nutrient sources. Overall, our findings suggest that during the pigmentation process in H. leucospilota, no significant changes were observed in the classical melanin pathway, while notable alterations were observed in their nutritional status. This study provides valuable insights into the regulatory mechanisms of pigmentation in marine organisms.
Assuntos
Holothuria , Pepinos-do-Mar , Humanos , Animais , Pepinos-do-Mar/genética , Holothuria/genética , Estado Nutricional , Transcriptoma , Cromatografia Líquida , Melaninas/genética , Espectrometria de Massas em Tandem , Perfilação da Expressão Gênica , Pigmentação/genética , MetabolomaRESUMO
The tropical sea cucumber Holothuria scabra is naturally found in the Indo-West Pacific. However, due to their commercial value, natural H. scabra populations have declined significantly in recent years, resulting in its status as an endangered species. Surveys of H. scabra resource pose a challenge due to its specific characteristics, such as sand-burrowing behavior. To overcome this problem, our study established a convenient and feasible method for assessing H. scabra resources using environmental DNA (eDNA) monitoring technology. First, H. scabra-specific TaqMan primers and probe were designed based on its cox1 gene, followed by the development of an eDNA monitoring method for H. scabra in two separate sea areas (Xuwen and Daya Bay). The method was subsequently employed to investigate the distribution of H. scabra and assess the effects of aquaculture stock enhancement through juvenile releasing in the Weizhou Island sea area. The H. scabra eDNA monitoring approach was found to be more appropriate and credible than traditional methods, and a positive impact of stocking on H. scabra populations was observed. In summary, this is the first report to quantify eDNA concentration in a Holothuroidea species, and it provides a convenient and accurate method for surveying H. scabra resources. This study introduces novel concepts for eDNA-based detection of endangered marine benthic animals and monitoring their population distribution and abundance.
Assuntos
DNA Ambiental , Holothuria , Pepinos-do-Mar , Animais , Holothuria/genética , Pepinos-do-Mar/genética , DNA Ambiental/genéticaRESUMO
Extracellular vesicles (EVs), carriers of molecular signals, are considered a critical link in maintaining homeostasis in mammals. Currently, there is growing interest in studying the role of EVs, including exosomes (subpopulation of EVs), in animals of other evolutionary levels, including marine invertebrates. We have studied the possibility of obtaining appropriate preparations of EVs from whole-body extract of holothuria Eupentacta fraudatrix using a standard combination of centrifugation and ultracentrifugation. However, the preparations were heavily polluted, which did not allow us to conclude that they contained vesicles. Subsequent purification by FLX gel filtration significantly reduced the pollution but did not increase vesicle concentration to a necessary level. To detect EVs presence in the body of holothurians, we used transmission electron microscopy of ultrathin sections. Late endosomes, producing the exosomes, were found in the cells of the coelom epithelium covering the gonad, digestive tube and respiratory tree, as well as in the parenchyma cells of these organs. The study of purified homogenates of these organs revealed vesicles (30-100 nm) morphologically corresponding to exosomes. Thus, we can say for sure that holothurian cells produce EVs including exosomes, which can be isolated from homogenates of visceral organs.
Assuntos
Exossomos , Vesículas Extracelulares , Holothuria , Pepinos-do-Mar , Animais , Evolução Biológica , Vesícula , MamíferosRESUMO
The sea cucumber Holothuria leucospilota, a nutritive and commercial marine species, has a high protein and low lipid content. To date, the mechanisms underlying gender determination and differentiation in sea cucumbers remain unclear. Identifying gender-specific molecular markers is an effective method of revealing the genetic basis of gender determination and differentiation. The inability to distinguish between male and female individuals causes reproductive efficiency to decline in aquaculture. In this study, we used the gonads of the sea cucumber H. leucospilota as samples to conduct the experiment. The differentially abundant metabolites (DAMs) detected by liquid chromatography-mass spectrometry were enriched in pathways associated with prolactin metabolism, insulin metabolism, hypoxia-inducible factor-1 signaling, and calcium signaling. At the transcriptome level, Illumina sequencing was performed on H. leucospilota, demonstrating that gender-specific expression genes were enriched in the retinoic acid-inducible gene I-like receptor signaling pathway, C-type lectin receptor signaling pathway, alpha-linolenic acid metabolism, and ether lipid metabolism by the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. By analyzing the common pathways between DAMs and differentially expressed genes, we found that gender-related genes of H. leucospilota were mostly enriched in the necroptosis pathway and the cysteine and methionine metabolism pathways. According to the common pathways, uch-sc1 and uch-sc2 are male-specific expression genes, and uch-sc3 and bhmt are female-specific expression genes at the mRNA level. These results provide information on gender differences in H. leucospilota.
Assuntos
Holothuria , Pepinos-do-Mar , Humanos , Animais , Feminino , Masculino , Transcriptoma , Metaboloma , Sinalização do CálcioRESUMO
The sea cucumber body wall was subjected to enzymatic hydrolysis using papain. The relationship between the enzyme concentration (1-5% w/w protein weight) and hydrolysis time (60-360 min) and the degree of hydrolysis (DH), yield, antioxidant activities, and antiproliferative activity in a HepG2 liver cancer cell line was determined. The surface response methodology showed that the optimum conditions for the enzymatic hydrolysis of sea cucumber were a hydrolysis time of 360 min and 4.3% papain. Under these conditions, a 12.1% yield, 74.52% DH, 89.74% DPPH scavenging activity, 74.92% ABTS scavenging activity, 39.42% H2O2 scavenging activity, 88.71% hydroxyl radical scavenging activity, and 9.89% HepG2 liver cancer cell viability were obtained. The hydrolysate was produced under optimum conditions and characterized in terms of its antiproliferative effect on the HepG2 liver cancer cell line.
Assuntos
Holothuria , Neoplasias Hepáticas , Pepinos-do-Mar , Animais , Humanos , Antioxidantes/farmacologia , Antioxidantes/química , Hidrólise , Pepinos-do-Mar/química , Papaína , Peróxido de Hidrogênio , Neoplasias Hepáticas/tratamento farmacológico , Linhagem Celular , Hidrolisados de Proteína/farmacologia , Hidrolisados de Proteína/químicaRESUMO
Bisphenols (BPs) are among emerging pollutants that have been frequently detected in different compartments of marine ecosystems and elicited great concern due to their potential toxicity to marine organisms. This work aimed to investigate the toxicity of bisphenol A (BPA) and bisphenol S (BPS) on oxidative stress markers, neurotoxicity and histopathological alterations in sea cucumbers (Holothuria poli). The results showed that exposure to 200 µg/L of BPA and BPS produced oxidative stress, neurotoxicity in the digestive tract and respiratory tree, and several types of histopathological lesions in tissues of the respiratory tree of the sea cucumber, posing a health hazard to this aquatic organism. In addition, BPA has greater effects than BPS on the generation of oxidative stress marked by the inductions of catalase (CAT), glutathione S-transferase (GST) and malondialdehyde (MDA) levels and neurotoxicity shown by the decrease in acetylcholinesterase activity (AChE). The respiratory tree of sea cucumbers might be an appropriate tissue for assessing CAT, MDA and AChE activity levels, which are reliable biomarkers that may be useful in marine biomonitoring studies. Evaluation of histopathological lesions of the respiratory tree suggests that BPA and BPS and their mixture cause various tissue alterations that may be associated with oxidative stress damage and neurotoxicity. In conclusion, this study showed that oxidative stress (CAT and MDA) and neurotoxicity (AChE) markers, as well as respiratory tree lesions, are sensitive biomarkers for the assessment of BPA and BPS toxicity in sea cucumbers.
Assuntos
Holothuria , Pepinos-do-Mar , Animais , Acetilcolinesterase , Ecossistema , BiomarcadoresRESUMO
The present work assess the bioconcentration kinetics of atenolol (ATN) and carbamazepine (CBZ) in common marine organisms including Holothuria tubulosa, Anemonia sulcata and Actinia equina under controlled laboratory conditions. CBZ exhibited higher uptake and excretion rates resulting higher bioconcentration factor (BCF) (41-537 L/kg for CBZ vs 7-50 L/kg for ATN) although both are below the limits established by the European Union (EU). The measured BCF using kinetic data showed some differences with those predicted using the concentrations measured at the steady-state, probably explained because the steady state was not ready reached. The animal-specific BCF followed the order of Holothuria tubulosa > Actinia equina > Anemonia sulcata for ATN while was the opposite for CBZ. The study highlighted between-tissues differences in the digestive tract and the body wall of the Holothuria tubulosa. The work presented is the first to model bioconcentration of ATN and CBZ in holothurian and anemone animal models.
Assuntos
Holothuria , Poluentes Químicos da Água , Animais , Bioacumulação , Atenolol , Organismos Aquáticos , Carbamazepina , Preparações Farmacêuticas , Poluentes Químicos da Água/toxicidade , Poluentes Químicos da Água/análiseRESUMO
The assessment of exposure to the antidepressant venlafaxine and its major metabolite o-desmethylvenlafaxine in Holothuria tubulosa, Anemonia sulcata and Actinia equina is proposed. A 28-day exposure experiment (10 µg/L day) followed by a 52-day depuration period was conducted. The accumulation shows a first-order kinetic process reaching an average concentration of 49,125/54342 ng/g dw in H. tubulosa and 64,810/93007 ng/g dw in A. sulcata. Venlafaxine is considered cumulative (BCF > 2000 L/kg dw) in H. tubulosa, A. sulcata and A. equina respectively; and o-desmethylvenlafaxine in A. sulcata. Organism-specific BCF generally followed the order A. sulcata > A. equina > H. tubulosa. The study revealed differences between tissues in metabolizing abilities in H. tubulosa this effect increases significantly with time in the digestive tract while it was negligible in the body wall. The results provide a description of venlafaxine and o-desmethylvenlafaxine accumulation in common and non-target organisms in the marine environment.