Characterization of myosin light chain in shrimp hemocytic phagocytosis.
Myosin light chain, a well-known cytoskeleton gene, regulates multiple processes that are involved in material transport, muscle shrink and cell division. However, its function in phagocytosis against invading pathogens in crustacean remains unknown. In this investigation, a myosin light chain gene was obtained from Marsupenaeus japonicus shrimp. The full-length cDNA of this gene was of 766 bp and an open reading frame (ORF) of 462 bp encoding a polypeptide of 153 amino acids. The myosin light chain protein was expressed in Escherichia coli and purified. Subsequently the specific antibody was raised using the purified GST fusion protein. As revealed by immuno-electron microscopy, the myosin light chain protein was only expressed in the dark bands of muscle. In the present study, the myosin light chain gene was up-regulated in the WSSV-resistant shrimp as revealed by real-time PCR and western blot. And the phagocytic percentage and phagocytic index using FITC-labeled Vibrio parahemolyticus were remarkably increased in the WSSV-resistant shrimp, suggesting that the myosin light chain protein was essential in hemocytic phagocytosis. On the other hand, RNAi assays indicated that the phagocytic percentage and phagocytic index were significantly decreased when the myosin light chain gene was silenced by sequence-specific siRNA. These findings suggested that myosin light chain protein was involved in the regulation of hemocytic phagocytosis of shrimp.
Han
Fang
F
College of Animal Science and Technology, Hunan Agricultural University, Changsha 410128, China.
Wang
Zhiyong
Z
Wang
Xiaoqing
X
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D016000:Cluster Analysis; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli; D006434:Hemocytes / Q000276:immunology*; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D018994:Myosin Light Chains / Q000235:genetics* / Q000276:immunology* / Q000648:ultrastructure; D016366:Open Reading Frames / Q000235:genetics; D033561:Penaeidae / Q000276:immunology*; D010587:Phagocytosis / Q000276:immunology*; D010802:Phylogeny; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014736:Vibrio parahaemolyticus
D017931:DNA Primers; D018076:DNA, Complementary; D018994:Myosin Light Chains