Proteomic analysis of differentially expressed proteins in Penaeus monodon hemocytes after Vibrio harveyi infection

Abstract

BackgroundViral and bacterial diseases can cause mass mortalities in commercial shrimp aquaculture. In contrast to studies on the antiviral response, the responses of shrimps to bacterial infections by high throughput techniques have been reported only at the transcriptional level and not at the translational level. In this study, a proteomic analysis of shrimp hemocytes to identify differentially expressed proteins in response to a luminous bacterium Vibrio harveyi was evaluated for its feasibility and is reported for the first time.ResultsThe two-dimensional gel electrophoresis (2-DE) patterns of the hemocyte proteins from the unchallenged and V. harveyi challenged shrimp, Penaeus monodon, at 24 and 48 h post infection were compared. From this, 27 differentially expressed protein spots, and a further 12 weakly to non-differentially regulated control spots, were selected for further analyses by the LC-ESI-MS/MS. The 21 differentially expressed proteins that could be identified by homologous annotation were comprised of proteins that are directly involved in the host defense responses, such as hemocyanin, prophenoloxidase, serine proteinase-like protein, heat shock protein 90 and alpha-2-macroglobulin, and those involved in signal transduction, such as the14-3-3 protein epsilon and calmodulin. Western blot analysis confirmed the up-regulation of hemocyanin expression upon bacterial infection. The expression of the selected proteins which were the representatives of the down-regulated proteins (the 14-3-3 protein epsilon and alpha-2-macroglobulin) and of the up-regulated proteins (hemocyanin) was further assessed at the transcription level using real-time RT-PCR.ConclusionsThis work suggests the usefulness of a proteomic approach to the study of shrimp immunity and revealed hemocyte proteins whose expression were up regulated upon V. harveyi infection such as hemocyanin, arginine kinase and down regulated such as alpha-2-macroglobulin, calmodulin and 14-3-3 protein epsilon. The information is useful for understanding the immune system of shrimp against pathogenic bacteria.

Cloning and characterization of the hemocyanin gene of prawn Macrobrachium nipponense

“…Recently, antibacterial peptides have been isolated and purified from C-terminus of hemocyanin from L. vannamei, Litopenaeus stylirostris [22], and P. leniusculus [23]. After infected by Vibrio harveyi, hemocyanin gene of Penaeus monodon expression was remarkably enhanced [6] . Our results showed that expression of MNHc mRNA in hepatopancreas was decreased at 3 h after prawns were challenged with pathogenic bacteria A. Hydrophila (Fig.…”

“…Prawns, like other invertebrates, lack adaptive immunity and they have to rely on innate immune systems for defensing and eliminating microorganisms [7,8]. Many non-specific molecules, including phenoloxidase, antimicrobial peptides, lysozyme and lectins, specifically hemocyanin, have been cloned and characterized in succession [1,5,6,9]. Up to date, however, there is no report about hemocyanin gene from M. nipponense.…”

“…As the main protein component of haemolymph, hemocyanin typically represents up to 90% of total amount of protein [1]. Except for its primary function as an oxygen-carrier, hemocyanin is a multi-functional protein involved in several physiological processes and immue defence such as phenoloxidase activity [2,3], hemolytic activity [4], anti-virus [5] and anti-bacteria [6]. The freshwater prawn Macrobrachium nipponense (M. nipponense) an economically important species, is a primary inland cultured species in China, Japan and Annam.…”

El camarón como una fuente de alérgenos

“…Su contenido alergénico puede alterarse por diversos factores, como infección por virus, estrés por hipoxia y dieta en los cultivos de camarón (20)(21)(22)(23). En hemocitos del camarón L. vannamei se identificaron varios grupos de proteínas cuya expresión es regulada por la infección con el virus del síndrome de Taura (Taura Syndrome Virus, TSV) (24).…”

“…En la especie P. monodon las infecciones por la bacteria Vibrio harveyi aumentan la expresión de algunas proteínas diferentes a las que son reguladas por el TSV en L. vannamei. Los camarones infectados con esta bacteria tienen aumento de la expresión de hemocianinas, arginina-cinasa, profelonixidasa, serina-proteasa y la proteína de choque térmico (22,25).…”