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Characterization of PmSpӓtzle 1 from the black tiger shrimp Peneaus monodon
Sittichai Boonrawd1, Ravi Mani2, Sirikwan Ponprateep3
1Center of Excellence for Molecular Biology and Genomics of Shrimp, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phyathai Road, Bangkok 10330, Thailand.
Abstract:
Spätzle is a signaling ligand in innate immune response that signals pathogenic infection via Toll receptor and Toll pathway into the cells for the synthesis of antimicrobial proteins. Herein, three PmSpӓtzle isoforms were identified in Penaeus monodon, namely PmSpz1, 2 and 3. The PmSpz1 was chosen for detailed study. The PmSpz1 gene was expressed in all nine tissues tested including the hemocytes, stomach, hepatopancreas, gill, lymphoid tissue, eyestalk, muscle, intestine and heart. Its expression was up-regulated upon white spot syndrome virus (WSSV) infection. Western blot analysis of hemolymph showed that the PmSpz1 mostly existed as a cleaved active form awaiting to activate the Toll pathway. Injection of a recombinant PmSpz1 rendered the shrimp less susceptible to the WSSV infection. Injection of a recombinant active form of PmSpz1 into a normal shrimp activated the synthesis of crustinPm1, crustinPm7, ALFPm3, penaeidin3 but not penaeidin5 indicating that the expression of all antimicrobial proteins but not penaeidin5 was under the regulation of Toll pathway.
Insights
This study identifies three Spätzle isoforms in shrimp, finding that PmSpätzle1 activates antimicrobial protein synthesis. This PmSpätzle1 enhances shrimp resistance to white spot syndrome virus (WSSV) infection.
Area of Science:
- Immunology
- Marine Biology
- Molecular Biology
Background:
- Spätzle is a key signaling ligand in innate immunity, crucial for antimicrobial protein synthesis via the Toll pathway.
- Understanding shrimp immune responses is vital for aquaculture and disease management.
Purpose of the Study:
- To identify and characterize Spätzle isoforms in Penaeus monodon.
- To investigate the role of PmSpätzle1 in the shrimp immune response, particularly against white spot syndrome virus (WSSV).
Main Methods:
- Identification and sequencing of PmSpätzle isoforms.
- Gene expression analysis across various shrimp tissues.
- Western blot to detect active PmSpätzle1.
- Experimental infection with WSSV and injection of recombinant PmSpätzle1.
Main Results:
- Three PmSpätzle isoforms (PmSpz1, PmSpz2, PmSpz3) were identified in Penaeus monodon.
- PmSpätzle1 gene expression was detected in all tested tissues and upregulated upon WSSV infection.
- Recombinant PmSpätzle1 injection increased shrimp resistance to WSSV and activated specific antimicrobial proteins (crustinPm1, crustinPm7, ALFPm3, penaeidin3) via the Toll pathway.
Conclusions:
- PmSpätzle1 is an important immune signaling molecule in shrimp.
- PmSpätzle1 plays a significant role in the Toll pathway-mediated innate immune response against WSSV.
- The study highlights the potential of PmSpätzle1 as a target for disease control strategies in shrimp aquaculture.