Demonstration of a true phenoloxidase activity and activation of a ProPO cascade in Pacific oyster, Crassostrea gigas (Thunberg) in vitro.
The prophenoloxidase (ProPO) system is the origin of melanin production and is considered to be an innate defence mechanism in invertebrates. In different bivalve species, phenoloxidase (PO) is present in the haemolymph as an inactive form of ProPO. The present study focuses on the Pacific adult oyster, Crassostrea gigas, an economically important bivalve species along French coasts. The results indicate that many factors may inhibit the PO-like activity. These include: phenylthiourea (PTU), sodium diethylthiocarbamate (DETC), beta-mercaptoethanol and tropolone, which repressed the spontaneous PO activity. The activation of PO-like activity in C. gigas acellular fraction by lipopolysaccharide (LPS) involved participation of other factors, including at least one serine protease. PO was present as proPO in the acellular fraction of haemolymph and haemocytes of C. gigas and could be activated by an exogenous protease (trypsin-N-tosyl-l-phenylalanine chloromethyl ketone) when used at 1 gL(-1). Treatment of acellular fractions with other modulators/activators namely LPS (1 gL(-1)), zymosan (0.6 gL(-1)) or laminarin (0.6 gL(-1)) also increased PO-like activity but to a less important way. The study demonstrated the evidence of a true phenoloxidase activity in Pacific oyster, C. gigas (Thunberg). The activation of a proPO system by non-self molecules suggests the role played by PO in vivo in the internal defence mechanisms. Understanding the activation of the ProPO system could enable the evaluation of the health of oyster stocks.
Hellio
Claire
C
School of Biological Sciences, University of Portsmouth, King Henry Building, Portsmouth PO1 2DY, UK.
Bado-Nilles
Anne
A
Gagnaire
Béatrice
B
Renault
Tristan
T
Thomas-Guyon
Hélène
H
D000818:Animals; D004156:Catechol Oxidase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D049896:Crassostrea / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D020536:Enzyme Activators / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000276:immunology*; D006458:Hemolymph / Q000187:drug effects / Q000201:enzymology*
D020536:Enzyme Activators; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; C038451:pro-phenoloxidase; D004156:Catechol Oxidase