RESUMEN
Effectsof microwave vacuum drying (MVD) on moisture migration, microstructure, and rehydration of sea cucumber were investigated in this paper. Vacuum condition avoided the exposure of sea cucumber to high temperature. Low-field nuclear magnetic resonance relaxation results revealed that the peaks of three water components in sea cucumber shifted to short relaxation time during MVD process, and the peak area of major water component-immobilized water-decreased significantly due to water evaporation. Magnetic resonance imaging found that the water in the internal layer of sea cucumber body wall was first removed due to the internal heating of microwave, and then the water in the outer layer. Higher microwave power could promote the moisture transfer motion during drying process, and shorten the drying time. Porous microstructure was observed by Cryo scanning electronic microscope images in sea cucumber dried with microwave power of 200 and 250 W, which might be responsible for high values of rehydration ratio and water holding capacity. High microwave power caused the increase of amino acids content, but had no significant effect on the change of saponins content. In addition, excellent prediction models of moisture ratio have been developed by partial least squares regression analysis based on transverse relaxation data, which proved the feasibility of low-field nuclear magnetic resonance to monitor moisture changes of sea cucumber during MVD process. PRACTICAL APPLICATION: Effects of microwave vacuum drying (MVD) on moisture migration, microstructure, and rehydration of sea cucumber were investigated. Understanding the impacts of MVD drying on water status, texture, and nutritional characteristics of sea cucumber is important to improve the processing quality of dried sea cucumber.
Asunto(s)
Fluidoterapia/métodos , Microondas , Pepinos de Mar/química , Agua/química , Animales , Desecación/métodos , Calefacción , Pepinos de Mar/fisiología , Pepinos de Mar/efectos de la radiación , VacioRESUMEN
The sea cucumber Apostichopus japonicus (Selenka)is a typical nocturnal echinoderm, which is believed to be almost completely dependent on light intensity for the regulation of endogenous rhythms. Under conditions of high light intensity, this species shows clear evidence of light avoidance behavior, seeking out shaded areas of reef in which to reside. In this study, we performed RNA-Seq analysis to examine the tentacle transcriptome of A. japonicus specimens that had been subjected to dark and light (5 min and 1 h) conditions. We specifically focused on detecting genes involved in opsin-based light perception, including opsins and members of phototransduction-related pathways. On the basis of comparisons with both vertebrate and invertebrate phototransduction pathways, we determined that components of two of the main metazoan phototransduction pathways were altered in response to illumination. Among the key phototransduction-related genes in tentacles, we identified retinol dehydrogenase, members of the dehydrogenase/reductase family, and myosin III, and also detected a pair of visual pigment-like receptors, peropsin and peropsin-like, the homologous genes of which are believed to have the same function but show opposite expression patterns in response to different light environments. In general, the up-regulation of key genes in sea cucumber exposed to illumination indicated that the tentacles can respond to differences in the light environment at the molecular level.
Asunto(s)
Estructuras Animales/metabolismo , Regulación de la Expresión Génica/efectos de la radiación , Fototransducción/genética , Pepinos de Mar/genética , Transcriptoma/efectos de la radiación , Estructuras Animales/efectos de la radiación , Animales , Perfilación de la Expresión Génica , Pepinos de Mar/efectos de la radiaciónRESUMEN
Sea cucumber body-wall melting occurs under certain circumstances. We have shown that apoptosis but not autolysis plays a critical role in the initial stage. However, it is still unclear how apoptosis is triggered in this process. In this study, we examined the levels of reactive oxygen species (ROS), the levels of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax) proteins, the depolarization of mitochondrial transmembrane potentials, and cytochrome c (Cyt c) release during sea cucumber melting induced by ultraviolet (UV) exposure. We also investigated the contribution of caspase in this process by injecting a pan-caspase inhibitor. Our data showed that UV exposure stimulates ROS production, dysfunction of mitochondria, and the release of Cyt c in sea cucumber coelomic fluid cells and body walls. We found a decrease of Bcl-2 and increase of Bax in the mitochondria after UV exposure. We also demonstrated that these changes are associated with elevated caspase-9 and -3 activity. Finally, our data showed that the inhibition of caspases-9 and -3 using an inhibitor suppresses UV-induced sea cucumber melting. These results suggest that apoptosis during sea cucumber melting is mediated by mitochondrial dysfunction and follows the activation of the caspase-signaling pathway. This study presents a novel insight into the mechanism of sea cucumber melting.
Asunto(s)
Caspasas/metabolismo , Pepinos de Mar/fisiología , Pepinos de Mar/efectos de la radiación , Clorometilcetonas de Aminoácidos/farmacología , Animales , Apoptosis/efectos de la radiación , Inhibidores de Caspasas/farmacología , Citocromos c/metabolismo , Potencial de la Membrana Mitocondrial/efectos de la radiación , Mitocondrias/metabolismo , Transporte de Proteínas/efectos de la radiación , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Pepinos de Mar/efectos de los fármacos , Rayos Ultravioleta , Proteína X Asociada a bcl-2/metabolismoRESUMEN
The autolysis of sea cucumber (Stichopus japonicus) was induced by ultraviolet (UV) irradiation, and the changes of microstructures of collagenous tissues and distributions of cathepsin L were investigated using histological and histochemical techniques. Intact collagen fibers in fresh S. japonicus dermis were disaggregated into collagen fibrils after UV stimuli. Cathepsin L was identified inside the surface of vacuoles in the fresh S. japonicus dermis cells. After the UV stimuli, the membranes of vacuoles and cells were fused together, and cathepsin L was released from cells and diffused into tissues. The density of cathepsin L was positively correlated with the speed and degree of autolysis in different layers of body wall. Our results revealed that lysosomal cathepsin L was released from cells in response to UV stimuli, which contacts and degrades the extracellular substrates such as collagen fibers, and thus participates in the autolysis of S. japonicus.
Asunto(s)
Autólisis/patología , Catepsina L/metabolismo , Colágeno/química , Pepinos de Mar/química , Rayos Ultravioleta , Animales , Colágeno/ultraestructura , Microscopía Electrónica de Rastreo , Pepinos de Mar/efectos de la radiaciónRESUMEN
Autolysis easily happens to sea cucumber (Stichopus japonicus, S. japonicus) for external stimulus like UV exposure causing heavy economic losses. Therefore, it is meaningful to reveal the mechanism of S. japonicas autolysis. In the present study, to examine the involvement of apoptosis induction in UVA-induced autolysis of S. japonicas, we investigated the biochemical events including the DNA fragmentation, caspase-3 activation, mitogen-activated protein kinases (MAPKs) phosphorylation and free radical formation. Substantial morphological changes such as intestine vomiting and dermatolysis were observed in S. japonicus during the incubation after 1-h UVA irradiation (10W/m(2)). The degradation of the structural proteins and enhancement of cathepsin L activity were also detected, suggesting the profound impact of proteolysis caused by the UVA irradiation even for 1h. Furthermore, the DNA fragmentation and specific activity of caspase-3 was increased up to 12h after UVA irradiation. The levels of phosphorylated p38 mitogen activated protein kinase (MAPK) and phosphorylated c-Jun.-N-terminal kinase (JNK) were significantly increased by the UVA irradiation for 1h. An electron spin resonance (ESR) analysis revealed that UVA enhanced the free radical formation in S. japonicas, even through we could not identify the attributed species. These results suggest that UVA-induced autolysis in S. japonicas at least partially involves the oxidative stress-sensitive apoptosis induction pathway. These data present a novel insight into the mechanisms of sea cucumber autolysis induced by external stress.
Asunto(s)
Apoptosis/efectos de la radiación , Pepinos de Mar/efectos de la radiación , Rayos Ultravioleta , Animales , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosforilación , Pepinos de Mar/citologíaRESUMEN
This report concerns physiological function of mycosporine-like amino acids (MAA) as an active defense against the photooxidative effects of sunlight in marine organisms. Mycosporine glycine (MG) is a representative member of MAA family and was found to effectively suppress various detrimental effects of the Type-II photosensitization in biological systems, such as inactivation of mitochondrial electron transport, lipid peroxidation of microsomes, hemolysis of erythrocytes and growth inhibition of Escherichia coli. The presence of MG in solutions of eosin Y or methylene blue resulted in a marked decrease in the level of singlet oxygen (1O2) produced by the sensitizers under illumination. The rate constant of 1O2 quenching by MG was determined to be 5.6 x 10(7) M(-1) s(-1) by the time-resolved 1O2 luminescence decay method, which is higher than, or at least comparable to, the values for 1O2 reaction of well-known quenchers such as 1,4-diazabicyclo[2,2,2]octane and furfuryl alcohol. The results suggest that MG probably together with some other active MAA may play an important role in protecting marine organisms against sunlight damage by eliminating 1O2 generated from certain endogenous photosensitizers.