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Synthesis and Action of Diapause Hormone of Bombyx mori.

家蚕滞育激素的合成及其作用。

基本信息

批准号：
03404008
负责人：
YAMASHITA Okitsugu
金额：
$ 19.39万
依托单位：
Nagoya University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (A)
财政年份：
1991
资助国家：
日本
起止时间：
1991 至 1993
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03404008/
关键词：
Diapause hormone Suboesophageal ganglion Diapause hormone precursor gene trehalase cDNA gene expression ovary diapause Bombyx mori 遺伝子発現神経ペプチド cDNA ペプチド合成トレハラ-ゼ

项目摘要

A neuropeptide, diapause hormone (DH) is responsible for induction of embryonic diapause of the silkworm, Bombyx mori. A new molecule having diapause inducing activity has been extracted from adult male heads and purified to the homogeneous state by HPLC.An apparent molecular weight of this peptide was 10,000, and the N-terminal 40 amino acid sequence was determined.Using the cDNA encoding diapause hormone precursor (DHP), the DHP gene was cloned, and its organization and nucleotide sequence were determined. The expression of this gene was clearly found in suboesophageal ganglion (SG) of the last instar larvae and pupae, and in pharate larvae which were incubated at high temperature. Histochemical studies using cDNA and anti-DH antibody clearly revealed that DH was synthesized in 12 neurosecretory cells of SG and released into circulation through corpus cardiacum-corpora allata.To estimate the action point of DH, we at first cloned a cDNA encoding ovary trehalase, and established a method for measurement of trehalase mRNA content in ovaries. The synthetic DH was injected into pharate adults in which ovarian development proceeds, or added to the medium for in vitro culture of ovaries. Both experiments have demonstrated that DH induces the expression of trehalase gene in ovaries, and this induction is inhibited by addition of actinomycin D, not cycloheximide. Thus, we concluded that DH expresses its function on developing ovaries to induce directly trehalase gene expression.

滞育激素（DH）是一种诱导家蚕胚胎滞育的神经肽。从雄性成虫头部提取了一个具有滞育诱导活性的新分子，经HPLC纯化至均一状态，其表观分子量为10，000，测定了N端40个氨基酸的序列，利用编码滞育激素前体（DHP）的cDNA克隆了DHP基因，并测定了其结构和核苷酸序列。该基因在末龄幼虫和蛹的食管下神经节（SG）以及高温培养的蚤状幼虫中均有表达。利用cDNA和抗DH抗体的组织化学研究表明，DH在SG的12个神经分泌细胞中合成，并通过心体-咽侧体释放到循环中。将合成的DH注射到卵巢发育进行的成体体内，或加入到卵巢体外培养的培养基中。这两个实验都表明，DH诱导海藻糖酶基因在卵巢中的表达，并且这种诱导被放线菌素D抑制，而不是放线菌酮。因此，我们认为DH在发育中的卵巢中直接诱导海藻糖酶基因的表达。