| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2023: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2022: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2021: ¥800,000 (Direct Cost: ¥800,000)
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| Outline of Annual Research Achievements |
The purpose of my research is the investigation of the novel function of the fibrinogen in the immune system. I established the fibrinogen knockout medaka which shows anemia with smaller number of red blood cells (RBCs) and more immature RBCs. I tried to use two gram-negative bacteria, Flavobacterium columnare and Aeromonas hydrophila, to infect the medaka (Oryzias latipes) through immersing. However, the expression level of several immune-related factors were not increased after infection in RT-qPCR result. Then, I changed the induction methods to the intraperitoneal injection. In this experiment, the lipopolysaccharide (LPS) which is derived from the gram-negative bacteria cell wall, was injected into medaka intraperitoneally. After injection, the immune response was successfully induced with the elevated expression of several pro-inflammatory cytokines, including IFN-gamma, TNF-alpha, hepcidin, etc. There's also significant difference between wildtype (WT) and fibrinogen gamma knockout (KO) medaka in IL-1 beta and complement C1q subunit C expression upon infection.
In order to investigate the role of fibrinogen and its interacting protein in these immune pathway, the immunoprecipitation WB and mass spectrometry was carried out using WT and KO medaka upon infection. Through the immunoprecipitation using anti-medaka fibrinogen gamma antibody, several possible interacting protein was detected including histone. In order to investigate the function of fibrinogen in interaction with the histone in immune system, further investigation is necessary.
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