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Continuous Production of Silkworm Polyvoltine Strain Eggs by Ideal Analogs of Diapause Hormone

滞育激素理想类似物连续生产蚕多化品系卵

基本信息

批准号：
05556008
负责人：
YAMASHITA Okitsugu
金额：
$ 9.66万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05556008/
关键词：
Diapause hormone Diapause hormone analog Diapause hormone gene gene expression neurosecretory cell diapause egg Bombyx mori 修飾ペプチド構造活性相関多化性系統

项目摘要

A neuropeptide hormone, diapause hormone (DH) is a 24 amino acid peptide amide which is responsible for induction of embryonic diapause of the silkworm, Bombyx mori. DH or its analog is a novel chemical agent to be useful for long term preservation of eggs of the polyvoltine strain of the silkworm. In order to develop some active analog of DH,we chemically synthesized more than 100 species of DH fragment analogs, and tested their diapause egg inducing activity. The result led con-clusions that the amidation of C terminus and the C-terminal pentapeptide sequence, FXPRL, are essential for expression of biological activity of DH.Thus, the C-terminal hexa peptide amide seemed to be an agonist of DH.To obtain a new DH-like peptide which is lipophilic and permeable through integment of silkworm pupae, we extracted DH active substances from male adult heads by using an organic solvent. The active materials were purified, and one of them was isolated. The isolated substance was a peptide with a MW of about 10K,and its N-terminal amino acid sequencing showed high content of Val, Ala andPro. The peptide was weak in biological activity, but acted as a protectant against DH degradation.DH gene was organized with 6 exons interrupted by 5 introns, and was expressed in 12 neurosecretory cells of suboesophageal ganglion. The incubation temperarture of eggs strictly affected the expression of the gene, which indicates that the environmental regulation of diapause is mediated through DH gene expression.The chemically synthesized DH or its fragments was injected into pupae, by which treatment diapause eggs were obtained. These eggs accumulated much amounts of sorbitol and alanine as the natural diapause eggs. Thus, the DH agents are promising chemical regulators for long term preservation of eggs of polyvoltine strains of the silkworm.

滞育激素（diapause hormone，DH）是一种由24个氨基酸组成的肽酰胺类激素，具有诱导家蚕胚胎滞育的作用。DH或其类似物是一种新型的化学药剂，可用于家蚕多化性品系的卵的长期保存。为了开发具有活性的DH类似物，我们化学合成了100多种DH片段类似物，并对其诱导滞育卵的活性进行了测定。结果表明，DH的C端酰胺化和C端五肽序列FXPRL是DH生物活性表达所必需的，因此C端六肽酰胺可能是DH的激动剂。对活性物质进行了纯化，并分离出其中一种。分离物为一分子量约为10 K的多肽，N端氨基酸序列分析表明，该多肽含有较高的瓦尔、Ala和Pro。DH基因由6个外显子和5个内含子组成，在食管下神经节的12个神经分泌细胞中均有表达。将化学合成的DH或其片段注射到蛹中，获得处理滞育卵。这些卵与自然滞育卵一样积累了大量的山梨醇和丙氨酸。因此，DH制剂是一种很有前途的家蚕多化性品系卵长期保存的化学调节剂。