The peroxinectin of white shrimp Litopenaeus vannamei is synthesised in the semi-granular and granular cells, and its transcription is up-regulated with Vibrio alginolyticus infection

Abstract

Peroxinectin mRNA expression in the different types of haemocytes of white shrimp Litopenaeus vannamei was studied by in situ hybridisation using digoxigenin-UTP-labelled riboprobes and reverse transcription-polyrnerase chain reaction (RT-PCR). Granular cells (GC) and a mixture of semi-granular cells (SGC) and hyaiine cells (HC) were separated by 70% Percoll gradient centrifugation. Peroxinectin was synthesised in both GC and the mixture of SGC-HC. An in situ hybridisation assay indicated that peroxinectin mRNA expression occurred in GC and SGC, but not in HC. Peroxinectin transcript up-regulated significantly, whereas haemocyte count decreased significantly at 6, 12 and 24 h post Vibrio alginolyticus-injection with slower restoration as compared to that of saline-injected shrimp. The RT-PCR assay indicated that peroxinectin exists extensively in several species of decapod crustaceans including L. vannamei, freshwater prawn Macrobrachium rosenbergii, common caridina Caridina pseudodenticulata, stone crab Thalamita crenata and mud crab Scylla serrata suggesting that this protein plays an important role in defence against pathogens. (C) 2005 Elsevier Ltd. All rights reserved.