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Published on: March 9, 2010
14-3-3ζ and 14-3-3ε are involved in innate immune responses in Pacific abalone (Haliotis discus hannai)
Rui Xue1, Dinglong Yang1, Yijing Han2
1Muping Coastal Environmental Research Station, Yantai Institute of Coastal Zone Research, Yantai Shandong, 264117, PR China; Center for Ocean Mega-science, Chinese Academy of Sciences, Qingdao Shandong, 266071, PR China; Research and Development Center for Efficient Utilization of Coastal Bioresources, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences, Yantai Shandong, 264003, PR China.
Insights
Pacific abalone 14-3-3ζ and 14-3-3ε proteins are crucial for innate immunity against Vibrio harveyi. Knockdown studies revealed their roles in regulating hemocyte immune responses, including apoptosis and phagocytosis.
Area of Science:
- * Molecular Biology
- * Immunology
- * Marine Biology
Background:
- * 14-3-3 proteins are vital for cellular processes, but their function in mollusks remains largely unexplored.
- * Pacific abalone (Haliotis discus hannai) possess two identified 14-3-3 genes, 14-3-3ζ and 14-3-3ε, with significant sequence similarity.
- * These genes are constitutively expressed in various abalone tissues, particularly the gills.
Purpose of the Study:
- * To investigate the roles of 14-3-3ζ and 14-3-3ε in the innate immune response of Pacific abalone.
- * To elucidate the specific functions of these proteins during Vibrio harveyi infection.
Main Methods:
- * Identification and characterization of 14-3-3ζ and 14-3-3ε cDNAs in Haliotis discus hannai.
- * Gene expression analysis in different tissues and under bacterial challenge.
- * Functional analysis using gene knockdown techniques (siRNA) to assess impacts on immune parameters.
- * Measurement of hemocyte activity, apoptosis, reactive oxygen species (ROS) production, and gene expression of immune-related factors.
Main Results:
- * Vibrio harveyi infection significantly induced hemocyte activity, apoptosis, ROS formation, and caspase 3 expression.
- * Knockdown of 14-3-3ζ suppressed apoptosis, ROS formation, and caspase 3 expression, while 14-3-3ε knockdown partially restored phagocytosis.
- * Silencing of either 14-3-3ζ or 14-3-3ε led to decreased expression of antimicrobial peptides and pro-inflammatory cytokines.
- * Reciprocal regulation was observed, with knockdown of one gene significantly increasing the expression of the other.
Conclusions:
- * 14-3-3ζ and 14-3-3ε play critical, albeit distinct, roles in the innate immune defense of Pacific abalone against Vibrio harveyi.
- * These proteins are involved in modulating hemocyte immune responses, including apoptosis, phagocytosis, and the expression of immune-related genes.
- * Further research into 14-3-3 proteins could offer insights into mollusk immunity and potential biotechnological applications.
Abstract:
The 14-3-3 proteins play important roles in various cellular processes by binding to different ligands, but little is known about these proteins in mollusks. In this study, two 14-3-3 cDNAs were identified from the Pacific abalone Haliotis discus hannai (designated 14-3-3ζ and 14-3-3ε), possessing 59.40% identity with each other. Both genes were predominantly expressed in the gills of unchallenged abalones, and their mRNA signals could also be detected in several other tissues, including the mantle, hepatopancreas and ovary. However, after Vibrio harveyi challenge, hemocytes were induced significantly (p < 0.01). Meanwhile, phagocytosis was inhibited, but apoptosis, reactive oxygen species formation, and caspase 3 expression were significantly induced (p < 0.01), and they were all suppressed with 14-3-3ζ knockdown (p < 0.01). The differences were that silencing 14-3-3ε reverted the decline in the phagocytic rate derived from bacterial infection, while ROS formation was not influenced significantly. In addition, the expression levels of several antimicrobial peptide and proinflammatory cytokine genes were also decreased with the silencing of 14-3-3 genes. However, with the knockdown of 14-3-3ζ, the expression of 14-3-3ε was further significantly increased (p < 0.01), and vice versa. Overall, our results suggested that 14-3-3ζ and 14-3-3ε should play important roles in innate immunity against V. harveyi infection.
