Endocrinological Study with the Brain of Non Mulberry Silkworm Antheraea Assama Using in Vitro Technique

The in vitro study with the brain of non mulberry silkworm showed that the brains collected from the second to fourth day of fifth instar larvae were suitable to survive in culture media as represented by the assay of aldolase activity. The soluble protein content in the brains kept in vitro, for 18 hours at dark was found to be lower than the same observed for the fresh brains. The native PAGE analysis of the soluble protein extract of the fresh and incubated brain collected from Antheraea assama revealed three bands for the former and two for the later. The band having the lowest mobility was not detected in the protein extract of the incubated brain. It suggested probable hydrolysis or migration of that particular protein during incubation. Absence of the soluble protein in the media used, emphasized the possibility of hydrolysis of protein due to action of protease as certain reports justified the need of soluble protein not only for the growth of insect cell in vitro but also to protect the cultured cells from the action of protease. However, as reported earlier for other insects, the migration of peptides from the brain tö the media was also confirmed with the help of native PAGE analysis of the media (after incubating 40 brains for 18 hours at 30°C at dark).