Integrated analysis of exoskeletal surface profile and chitin-related gene expression on Macrophthalmus japonicus mud crabs exposed to hexabromocyclododecane.

Hexabromocyclododecanes (HBCDs), widely used brominated flame retardants, easily accumulate in aquatic organisms such as Macrophthalmus japonicus crabs, which inhabit tidal flat sediments. To analyze the effects of HBCD exposure in chitin-formed exoskeleton, we investigated molecular responses of chitin-related genes as well as physical changes of the exoskeletal surface form as a new biological end-point on M. japonicus. The expression patterns of chitin biosynthesis-, modification-, and degradation-related genes in the gills and hepatopancreases of M. japonicus were also analyzed. Additionally, the survivability and exoskeleton surface profiles of M. japonicus crabs were evaluated. M. japonicus chitin synthase expression was significantly downregulated, whereas that of the chitinase transcript was significantly upregulated upon exposure to all HBCD concentrations on day 7. Contrastingly, the gene expression of chitin deacetylase 1 significantly increased upon exposure to all HBCD concentrations on day 1, and this increase was significantly elevated on day 4. The expression of chitin deacetylase 1 was dose-dependent. Additionally, decreased survival and exoskeleton surface profile changes were observed in M. japonicus crabs exposed to all HBCD concentrations. These results suggest that exposure to HBCD induces changes in the synthesis, modification, and degradation of chitin, a pivotal component of the cuticular exoskeleton, and may disrupt the exoskeletal surface structure in M. japonicus crabs.

Changes in exoskeleton surface roughness and expression of chitinase genes in mud crab Macrophthalmus japonicus following heavy metal differences of estuary.

Risk assessment of heavy metals is important for the health evaluation of inhabiting species in aquatic ecosystem. This study investigated whether chitin exoskeleton of mud crab Macrophthalmus japonicus is affected by heavy metals in estuary sediments in Korea. We compared heavy metal concentrations and analyzed the expression of M. japonicus chitinase genes, which play the crucial role in the formation of chitin exoskeleton. Concentrations of heavy metals were highly observed in crab body inhabiting Hampyeong among estuarine sites. High expressions of chitinase 1 were observed in crab gill and hepatopancreas from Myodo, which is the site with the lowest concentration of heavy metal in crab body. The surface roughness of the exoskeleton decreased with the increased concentration of heavy metals accumulated in the crab body. These results suggest that the total bioconcentration of heavy metals in crabs affected the expression of chitinase genes and changes in the exoskeleton surface roughness.

Changes of exoskeleton surface roughness and expression of crucial participation genes for chitin formation and digestion in the mud crab (Macrophthalmus japonicus) following the antifouling biocide irgarol.

Irgarol is a common antifoulant present in coastal sediment. The mud crab Macrophthalmus japonicus is one of the most abundant of the macrobenthos in the costal environment, and its exoskeleton has a protective function against various environmental threats. We evaluated the effects of irgarol toxicity on the exoskeleton of M. japonicus, which is the outer layer facing the environment. We analyzed transcriptional expression of exoskeleton, molting, and proteolysis-related genes in the gill and hepatopancreas of these exposed M. japonicus. In addition, changes in survival and exoskeleton surface characteristics were investigated. In the hepatopancreas, mRNA expression of chitinase 1 (Mj-chi1), chitinase 4 (Mj-chi4), and chitinase 5 (Mj-chi5) increased in M. japonicus exposed to all concentrations of irgarol. Mj-chi1 and Mj-chi4 expressions from 1 to 10µgL(-1) were dose- and time-dependent. Ecdysteroid receptor (Mj-EcR), trypsin (Mj-Tryp), and serine proteinase (Mj-SP) in the hepatopancreas were upregulated in response to different exposure levels of irgarol at day 1, 4, or 7. In contrast, gill Mj-chi5, Mj-Tryp, and Mj-SP exhibited late upregulated responses to 10µgL(-1) irgarol compared to the control at day 7. Mj-chi1 showed early upregulation upon exposure to 10µgL(-1) irgarol and Mj-chi4 showed no changes in transcription in the gill. Gill Mj-EcR presented generally downregulated expression patterns. In addition, decreased survival and change of exoskeleton surface roughness were observed in M. japonicus exposed to the three concentrations of irgarol. These results suggest that exposure to irgarol induces changes in the exoskeleton, molting, and proteolysis metabolism of M. japonicus.