カイコガ脳-側心体-アラタ体の培養と羽化ホルモン活性

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  • Cultivation of the brain-corpora cardiaca-corpora allata complex of the silkworm, Bombyx mori and eclosion hormone activity of the cultured organs and medium.
  • カイコガ脳‐側心体‐アラタ体の培養と羽化ホルモン活性
  • カイコガ ノウ ソクシンタイ アラタタイ ノ バイヨウ ト ウカ ホルモン カ

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In order to clarify the regulation mechanism for eclosion hormone synthesis and secretion, fundamental culture conditions for brain-corpora Cardiaca-corpora allata complexes (Br-CC-CA complexes) of the silkworm, Bombyx mori, were established. When 10 brains were placed in 3ml of CSM-2F medium, eclosion hormone activity was detected throughout the cultured periods. On the 6th day of the cultivation, the positive response for Bombyx eclosion hormone assay from the cultured brains was about 60%, and thereafter the activity decreased with the culture. Eclosion hormone activity was first detectable from the cultured medium 6 days after the cultivation. The highest eclosion hormone titer in the cultured medium was 22.5 units. It was concluded that culture conditions were not appropriate for the hormone synthesis since the hormone titer in brains and culture media was extremely low compared with those in vivo. Therefore, the culture conditions were modified for using Br-CC-CA complexes. It seemed suitable to culture 10 Br-CC-CA complexes together in 1ml of CSM-2F medium with a gentle stream of air at 25±1°C in 16 L-8 D. In these conditions, an increase of about 2-fold in eclosion hormone titer was observed in a brain during 12 day culture periods. Although the eclosion hormone activity was not detected in CC-CA and media at the initiation of culture, hormone activity in CC-CA could be detected 4 days after onset of the cultivation. Furthermore, the hormone titer in CC-CA became 2 units after 12 days cultivation. On the other hand, 55 units of hormone titer was detected in 1ml of the medium 12 days after onset of cultivation, indicating that 5.5 units of hormone were released from 1 Br-CC-CA complex. Eclosion hormone titer detected from cultured Br-CC-CA complexes and media and expressed as the titer from a single Br-CC-CA complex coincided well with the hormone titer present in 1 intact animal. Morphological changes of brain and CA were scarcely observed during 16 days culture periods in the CSM-2F medium. These results indicate that the synthesis and secretion of eclosion hormone occurred during the culture periods.

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