Hemocyanin in the exoskeleton of crustaceans: enzymatic properties and immunolocalization

Adachi, Kohsuke; Endo, Hideki; Watanabe, Toshiki; Nishi­oka, Takaaki; Hirata, Takashi

doi:10.1111/j.1600-0749.2005.00217.x

Cited by 64 publications

(55 citation statements)

References 23 publications

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“…The arthropod cuticle was considered traditionally to be an inert supporting exoskeleton, a protective barrier against microorganisms and an exchange surface (Adachi et al 2005). However, recent evidence suggests that the arthropod cuticle also contributes to various biochemical and physiological processes ( Willis 1999;Willis et al 2005).…”

Section: Discussionmentioning

confidence: 99%

Locating the barnacle settlement pheromone: spatial and ontogenetic expression of the settlement-inducing protein complex ofBalanus amphitrite

2006

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Barnacles are prominent members of hard substratum benthic communities and their study has been important to advances in experimental ecology and contemporary ecological theory. Having recently characterized the cue to gregarious settlement of Balanus amphitrite, the settlement-inducing protein complex (SIPC), we use two polyclonal antibodies to examine the tissue distribution and ontogenetic expression of this glycoprotein. These antibodies were raised against two separate peptides located near the N-and C-termini of the SIPC and were used to detect the glycoprotein by western blotting and immunohistochemistry. By in situ hybridization we also show that the SIPC mRNA co-occurs with the expressed glycoprotein in the cuticles of both nauplius and cypris larval stages and the adult. In the larvae, the SIPC is expressed most strongly in the mouthparts and the hindgut of the stage 2 nauplius and in the thoracopods, antennules and bivalved carapace of the cyprid. In adult B. amphitrite, the expressed SIPC is present in protein extracts of the shell and in all organs that are lined by cuticular tissues. We suggest that the SIPC is produced by the epidermal cells that secrete the cuticle and discuss these observations with regard to earlier studies and the role of the SIPC as a contact pheromone.

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Section: Discussionmentioning

confidence: 99%

Locating the barnacle settlement pheromone: spatial and ontogenetic expression of the settlement-inducing protein complex ofBalanus amphitrite

2006

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“…Arthropod cuticle is not solely a physical barrier to the environment but can contribute to critical processes such as immune responses and cross-linking for cuticle hardening and calcification of exoskeleton (Ashida and Brey, 1995;Sugumaran and Nelson, 1998;Adachi et al, 2005;Suderman et al, 2006;Glazer et al, 2013). In crustaceans, cuticle tearing leaks hemolymph and releases proteins, which coagulate sealing the wound and preventing infection (Iwanaga et al, 1998;Li et al, 2002;Jiravanichpaisal et al, 2006).…”

Section: Discussionmentioning

confidence: 99%

Barnacle biology before, during and after settlement and metamorphosis: a study of the interface

Essock‐Burns

Gohad

Orihuela

et al. 2016

Journal of Experimental Biology

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Mobile barnacle cypris larvae settle and metamorphose, transitioning to sessile juveniles with morphology and growth similar to that of adults. Because biofilms exist on immersed surfaces on which they attach, barnacles must interact with bacteria during initial attachment and subsequent growth. The objective of this study was to characterize the developing interface of the barnacle and substratum during this key developmental transition to inform potential mechanisms that promote attachment. The interface was characterized using confocal microscopy and fluorescent dyes to identify morphological and chemical changes to the interface and the status of bacteria present as a function of barnacle developmental stage. Staining revealed patchy material containing proteins and nucleic acids, reactive oxygen species amidst developing cuticle, and changes in bacteria viability at the developing interface. We found that as barnacles metamorphose from the cyprid to juvenile stage, proteinaceous materials with the appearance of coagulated liquid were released into and remained at the interface. It stained positive for proteins, including phosphoprotein, as well as nucleic acids. Regions of the developing cuticle and the patchy material itself stained for reactive oxygen species. Bacteria were absent until the cyprid was firmly attached, but populations died as barnacle development progressed. The oxidative environment may contribute to the cytotoxicity observed for bacteria and has the potential for oxidative crosslinking of cuticle and proteinaceous materials at the interface.

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“…Hemocyanin can also be converted into phenoloxidase (Adachi et al, 2005;Adachi et al, 2001;Decker and Jaenicke, 2004). In addition, Adachi and colleagues (Adachi et al, 2005) identified hemocyanins in the cuticle of the shrimp Penaeus japonicus, and demonstrated the in vitro phenoloxidase activity of the enzyme. These authors further suggested that cuticular hemocyanin functions as a sclerotizing agent and/or an innate immunity factor.…”

Section: Introductionmentioning

confidence: 99%

“…These precursors are secreted into the hemolymph, where they can be activated by specific proteinases or can be deposited in the cuticle, where they are activated on site (Söderhäll and Cerenius, 1998). Crustacean hemocyanins are expressed in the hepatopancreas of several species (Adachi et al, 2005;Durstewitz and Terwilliger, 1997;van Holde and Miller, 1995) and are secreted to the hemolymph, where they occur as large extracellular multi-subunit molecules (van Holde and Miller, 1995). Hemocyanin can also be converted into phenoloxidase (Adachi et al, 2005;Adachi et al, 2001;Decker and Jaenicke, 2004).…”

Section: Introductionmentioning

confidence: 99%

“…Crustacean hemocyanins are expressed in the hepatopancreas of several species (Adachi et al, 2005;Durstewitz and Terwilliger, 1997;van Holde and Miller, 1995) and are secreted to the hemolymph, where they occur as large extracellular multi-subunit molecules (van Holde and Miller, 1995). Hemocyanin can also be converted into phenoloxidase (Adachi et al, 2005;Adachi et al, 2001;Decker and Jaenicke, 2004). In addition, Adachi and colleagues (Adachi et al, 2005) identified hemocyanins in the cuticle of the shrimp Penaeus japonicus, and demonstrated the in vitro phenoloxidase activity of the enzyme.…”

Section: Introductionmentioning

confidence: 99%

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Hemocyanin with phenoloxidase activity in the chitin matrix of the crayfish gastrolith

Glazer

Tom

Weil

et al. 2013

Journal of Experimental Biology

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SUMMARYGastroliths are transient extracellular calcium deposits formed by the crayfish Cherax quadricarinatus von Martens on both sides of the stomach wall during pre-molt. Gastroliths are made of a rigid chitinous organic matrix, constructed as sclerotized chitin-protein microfibrils within which calcium carbonate is deposited. Although gastroliths share many characteristics with the exoskeleton, they are simpler in structure and relatively homogeneous in composition, making them an excellent cuticle-like model for the study of cuticular proteins. In searching for molt-related proteins involved in gastrolith formation, two integrated approaches were employed, namely the isolation and mass spectrometric analysis of proteins from the gastrolith matrix, and 454-sequencing of mRNAs from both the gastrolith-forming and sub-cuticular epithelia. SDS-PAGE separation of gastrolith proteins revealed a set of bands at apparent molecular masses of 75-85kDa; mass spectrometry data matched peptide sequences from the deduced amino acid sequences of seven hemocyanin transcripts. This assignment was then examined by immunoblot analysis using anti-hemocyanin antibodies, also used to determine the spatial distribution of the proteins in situ. Apart from contributing to oxygen transport, crustacean hemocyanins were previously suggested to be involved in several aspects of the molt cycle, including hardening of the new post-molt exoskeleton via phenoloxidation. The phenoloxidase activity of gastrolith hemocyanins was demonstrated. It was also noted that hemocyanin transcript expression during pre-molt was specific to the hepatopancreas. Our results thus reflect a set of functionally versatile proteins, expressed in a remote metabolic tissue and dispersed via the hemolymph to perform different roles in various organs and structures.

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Hemocyanin in the exoskeleton of crustaceans: enzymatic properties and immunolocalization

Cited by 64 publications

References 23 publications

Locating the barnacle settlement pheromone: spatial and ontogenetic expression of the settlement-inducing protein complex ofBalanus amphitrite

Locating the barnacle settlement pheromone: spatial and ontogenetic expression of the settlement-inducing protein complex ofBalanus amphitrite

Barnacle biology before, during and after settlement and metamorphosis: a study of the interface

Hemocyanin with phenoloxidase activity in the chitin matrix of the crayfish gastrolith

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