Joint study on the mechanisms of reproduction in crustacea

甲壳类动物繁殖机制的联合研究

基本信息

批准号：
07044181
负责人：
NAGASAWA Hiromichi
金额：
$ 2.69万
依托单位：
Ocean Research Institute, University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

The endocrinological mechanisms of reproduction, especially vitellogenesis, in crustaceans have been investigated. It has been estimated that neuropeptides in the sinus gland inhibit vitellogenesis. We first extracted and purified seven structurally related neuropeptides from the sinus gland, and determined their whole amino acid sequences. They were designated as Pej-SGP-I--VII.All these peptides except for IV showed hyperglycemic acitivity, while IV had an activity to inhibit ecdysone release by Y-organ in in vitro culture system. These peptides were examined for vitellogenesis-inhibiting activity by using in vitro culture of pieces of developing ovary from the prawn Penaeus semisulcatus. The effects of these peptides on protein synthesis was examined by incorporation of ^<35>S-methionine into proteins in the ovarian pieces. The results showed that all the peptides except for IV inhibited protein synthesis. The inhibition was not confined to a specific protein but to all proteins. Ex … More periments of incorporation of ^3H-uridine showed that the inhibition occurred at the transcription level. By contrast, Pej-SGP-IV,red pigement concentrating hormone, pigment-dispersing hormone, 20-hydroxyecdysone or methyl farnesoate did not show any inhibiting or stimulating activity, suggesting the specific inhibition of six peptides.There has been no information about the structure of yolk protein vitellin or its precursor vitellogenin in crustaceans. First, vitellin was purified from the ovary of P.semisulcatus. By SDS-PAGE,it was separated into mainly 4 subunits, 160,120,96, and 80kDa. Blotted subunit proteins were sequenced and it was found that the N-terminal sequence of larger two were identical and that of smaller two were also identical. But the sequences between the two groups were different. Internal sequences were analyzed after digestion of native vitellin with lysyl endopeptidase and sebsequent separation of the digested fragments. Thus, we have obtained a lot of partial sequences. Using these peptide sequence information, we are now cloning cDNA encoding vitellogenin to know its full sequence. Less

甲壳类动物的生殖内分泌机制，特别是卵黄发生，据估计，窦腺中的神经肽抑制卵黄发生。我们首先从窦腺中提取和纯化了七种结构相关的神经肽，并确定了它们的完整氨基酸序列。它们被命名为Pej-SGP-I--VII。除IV外，所有这些肽均表现出高血糖活性，而IV具有抑制Y器官释放蜕皮激素的活性通过使用半沟对虾发育中的卵巢碎片的体外培养来检查这些肽的卵黄生成抑制活性。通过掺入^ 35 S-来检查这些肽对蛋白质合成的影响。结果表明，除 IV 之外的所有肽都抑制蛋白质合成，这种抑制并不局限于特定蛋白质，而是针对所有蛋白质。掺入^3H-尿苷的实验表明抑制发生在转录水平，相比之下，Pej-SGP-IV、红色素浓缩激素、色素分散激素、20-羟基蜕皮酮或法尼索酸甲酯没有表现出任何抑制或刺激作用。活性，表明六种肽具有特异性抑制作用。目前还没有关于甲壳类动物中卵黄蛋白卵黄蛋白或其前体卵黄原蛋白结构的信息。从P.semisulcatus卵巢中纯化得到卵黄蛋白，主要分为160、120、96和80kDa 4个亚基，对印迹亚基蛋白进行测序，发现其中较大的两个亚基的N端序列为。相同，较小的两个也相同，但用赖氨酰消化天然卵黄蛋白后，分析了两组之间的序列不同。内肽酶和随后的消化片段的分离因此，利用这些肽序列信息，我们现在克隆编码卵黄原蛋白的cDNA以了解其完整序列。

项目成果

期刊论文数量（24）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

長澤寛道(分担執筆): "昆虫の生化学・分子生物学" 名古屋大学出版会, 498 (1995)

Hiromichi Nagasawa（撰稿人）：“昆虫的生物化学和分子生物学”名古屋大学出版社，498（1995）

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Tsuyoshi Ohira et al.: "Cloning and sequence analysis of a cDNA encoding a crustacean hyperglycemic hormone from the Kuruma prawn Penaeus japonicus" Mol.Mar.Biol.Biotech.(in press). (1997)

Tsuyoshi Ohira 等人：“编码来自日本对虾甲壳动物高血糖激素的 cDNA 的克隆和序列分析”Mol.Mar.Biol.Biotech.（出版中）。

DOI：
发表时间：
期刊：
影响因子：
0
作者：
通讯作者：

Hiromichi Nagasawa,Wei-jun Yang,Hidetsugu Shimizu,Katsumi Aida,Hibiki Tsutsumi,Akiko Terauchi and Haruyuki Sonode: "Isolation and amino acid sequencemof amolt-i inhibiting hormone from the American crayfish,procam barus clarkii" Biosci・Biotech. Biochem.(印

Hiromichi Nagasawa、Wei-jun Yang、Hidetsugu Shimizu、Katsumi Aida、Hibiki Tsutsumi、Akiko Terauchi 和 Haruyuki Sonode：“来自美国克氏小龙虾的 amolt-i 抑制激素的分离和氨基酸序列”Biosci·Biochem。（标记