| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1990: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1989: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Research Abstract |
After confirming the existence of immunoglobulin (IgM) in fish eggs, following experiments were undertaken.
1. Assay systems for IgM of carp and rainbow trout were made. Rabbit was immunized with purified IgM. From the antiserum, IgG was purified and was biotinized. Using the IgG and biotinized IgG, enzyme immunoassay (EIA) system by biotin-avidin technique was made.
2. Serum IgM levels were estimated using EIA systems, with special reference to sexual maturation. In goldfish, IgM increased in the spring spawning season related with increase of water temperature. On the other hand, the IgM levels were rather low in the winter spawning season in rainbow trout.
3. Egg IgM in carp was analyzed. The supernatant of homogenized eggs was fractionated by gel filtration using HPLC. Proteins in each fraction was separated by SDS-PAGE and transferred on nito-cellulose membrane. The membrane was immuno-stained using biotinized anti-IgM IgG. Retention time of fractions containing IgM was longer than that of serum IgM, showing lower molecular weight. This lower mw was caused by smaller H-chain. The significance of the molecular differences in the defense mechanisms of eggs and fry are unknown.
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