Observation of Immune Gene Expression and Separated Haemocytes of Pacific White Shrimp, Litopenaeus vannamei
Date Issued
2015
Date
2015
Author(s)
Yang, Chih-Chiu
Abstract
Haemocyte of Shrimp played an important role in immune system; all of immune response of shrimp was active by haemocyte. In this study, L. vannamei haemocytes were firstly grouped into three different types and the immune related genes expression in grouped haemocytes were estimated. Those buffers include anticoagulant buffer and sheath fluid that was used by flow cytometery sorting system were modified according different environment of shrimp. White shrimp cultured in 10‰, 20‰ and 30% salinity water, osmolality are 507±43.89 mOsm/kg,779±29.96 mOsm/kg and 1136.67±98.17 mOsm/kg. According to those results, modified osmolality of buffers are 1200 mOsm/kg. Diverse haemocytes from Pacific white shrimp Litopenaeus vannamei were spread by flow cytometer sorting system. Using the two commonly flow cytometric parameters FSC and SSC, the haemocytes could be divided into three populations. Microscopy observation of L. vannamei hemocytes in anticoagulant buffer revealed three morphologically distinct cell types designated as granular cell, hyaline cell and semigranular cell. Immune genes, which includes prophenoloxidase (proPO), lipopolysaccharide--glucan binding protein (LGBP), peroxinectin, crustin, lysozyme, penaeid-3a and transglutaminase (TGase), expressed from different haemocyte were analysed by quantitative real time PCR (qPCR). Results from the mRNA expression were estimated by relative level of each gene to -actin gene. Finally, the seven genes could be grouped by their dominant expression sites. ProPO, LGBP and peroxinectin were highly expressed in granular cells, while LGBP, crustin, lysozyme and P-3a were highly expressed in semigranular cells and TGase was highly expressed in hyaline cells. In this study, L. vannamei haemocytes were firstly grouped into three different types and the immune related genes expression in grouped haemocytes were estimated.
Subjects
white shrimp
haemocyte
immune gene
Type
thesis
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