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Updated: Apr 27, 2026
Evaluation of Caspase Activation to Assess Innate Immune Cell Death
Published on: January 20, 2023
Three representative subtypes of caspase in miiuy croaker: genomic organization, evolution and immune responses to
Liping Ren1, Rixin Wang1, Tianjun Xu1
1Laboratory of Fish Biogenetics & Immune Evolution, College of Marine Science, Zhejiang Ocean University, Zhoushan 316022, China.
Abstract:
Caspase proteins are intracellular proteases which function as initiators and effectors of apoptosis. According their difference of functions, the caspases can be divided into apoptosis related caspases and inflammatory mediator, and the former included apoptosis activator and apoptosis executioner. In this study, three different subtype caspases (caspase1, caspase3 and caspase9) from Miichthys miiuy miiuy croaker were analyzed. The caspase1 belongs to the inflammatory mediator, caspase3 belongs to apoptosis executioner, and caspase9 belongs to apoptosis activator. Miichthys miiuy caspase1 (Mmcaspase1) and Mmcaspase9 exhibited three conserved domains including a CARD, a large subunit p20 and a small subunit p10. The Mmcaspase3 encoded two conserved domains including a large subunit p20 and a small subunit p10. Mmcaspase3 and Mmcaspase9 contained a histidine active sequence and a cysteine active sequence. However, Mmcaspase1 only contained the cysteine active sequence. The real-time PCR (RT-PCR) analysis showed these three caspases were expressed constitutively in all examined tissues in miiuy croaker, although the expression levels varied from tissue to tissue. Expression analysis showed that Mmcaspase1 was up-regulated obviously in liver, spleen and kidney, and indicated its positive role in response to Vibrio anguillarum infection, but Mmcaspase3 and Mmcaspase9 showed different expression pattern in liver, spleen and kidney, its showed that different subtypes of caspase having different immune response mechanisms. These results revealed that the organs adjusted the expressions of these three genes with the infected of pathogens, suggesting the immunoprotection of these genes.
Insights
This study analyzes three caspases in Miichthys miiuy croaker, revealing distinct roles in immune responses. Caspase-1 mediates inflammation, while caspase-3 and caspase-9 are involved in apoptosis, showing varied tissue expression and pathogen response.
Area of Science:
- Molecular Biology
- Immunology
- Aquatic Animal Health
Background:
- Caspase proteins are intracellular proteases crucial for apoptosis initiation and execution.
- Caspases are broadly classified into apoptosis-related (activators, executioners) and inflammatory mediators.
- Understanding fish caspase functions is vital for aquaculture health and disease management.
Purpose of the Study:
- To analyze the structure and expression of caspase-1, caspase-3, and caspase-9 in Miichthys miiuy croaker.
- To investigate the differential roles of these caspases in immune responses, particularly against Vibrio anguillarum infection.
- To elucidate the immunoprotective mechanisms mediated by these caspases in fish.
Main Methods:
- Bioinformatic analysis of conserved domains in Miichthys miiuy caspase genes (Mmcaspase1, Mmcaspase3, Mmcaspase9).
- Identification of active site sequences (histidine, cysteine) within the analyzed caspases.
- Real-time quantitative PCR (RT-PCR) to assess constitutive and infection-induced gene expression across various tissues.
Main Results:
- Mmcaspase1 and Mmcaspase9 possess three conserved domains (CARD, p20, p10), while Mmcaspase3 has two (p20, p10).
- Mmcaspase3 and Mmcaspase9 contain histidine and cysteine active sites; Mmcaspase1 contains only a cysteine active site.
- All three caspases are constitutively expressed, with Mmcaspase1 significantly upregulated in response to Vibrio anguillarum infection in liver, spleen, and kidney, indicating distinct immune roles.
Conclusions:
- Miichthys miiuy croaker caspases exhibit structural and functional diversity, reflecting specialized roles in apoptosis and inflammation.
- Differential expression patterns suggest unique immune response mechanisms for each caspase subtype against pathogens.
- Organ-specific regulation of caspase gene expression highlights their critical role in fish immunoprotection.
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