Lymnaea project - elucidation of mechanisms of molluscan shell formation

Lymnaea 项目 - 阐明软体动物壳形成的机制

• 批准号: 11440153
• 负责人: ENDO Kazuyoshi
• 金额: $ 8.58万
• 依托单位: University of Tsukuba (2002)The University of Tokyo (1999-2001)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11440153/
• 关键词: shell formation; morphogenesis; biomineralization; molluscs; Lymnaea; shell proteins; Hox genes; developmental genetics; 貝殼タンパク質; バイオシネラリゼーション; MSP-1; 殻内タンパク質

In order to obtain basic insight into developmental genetics of the pulmonate snail Lymnaea, we have conducted histological observations at various developmental stages. We have also prepared cDNA libraries for mRNA extracted from embryos of both before and after formation of the protochonch, and from adult mantle, muscle, and midgut gland tissues. Shell matrix proteins were analyzed using SDS-PAGE, and three prominent bands were separated. Subtraction cloning for the mantle specific genes using the digestive gland as a reference, resulted in isolation of a gene fragment encoding a member of P-type ATPases, as well as a fragment for a transcription factor containing a Zn-finger domain. So as to reverse genetically study functions of those gene products, basic techniques for microinjections and RNA interference for this species are now being developed. Concerning the shell matrix proteins of marine bivalves, we have isolated an extremely acidic protein, named Aspein, with 60% Asp residues, from the pearl oyster Pinctada fucata. The transcript for Aspein is shown to be specific to the calcitic prismatic layer part of the shell, suggesting that Aspein is involved in prismatic architecture formation and/or calcite precipitation.

为了获得基本的了解肺螺的发育遗传学，我们进行了组织学观察，在不同的发展阶段。我们还准备了cDNA文库的mRNA提取胚胎的前和后形成的原软骨，并从成人外套膜，肌肉和中肠腺组织。使用SDS-PAGE分析壳基质蛋白，并且分离出三个突出条带。使用消化腺作为参考的外套膜特异性基因的差减克隆，导致分离编码P型ATP酶的成员的基因片段，以及含有锌指结构域的转录因子的片段。为了从遗传学上研究这些基因产物的功能，目前正在开发用于该物种的显微注射和RNA干扰的基本技术。关于海洋双壳类的壳基质蛋白，我们从合浦珠母贝（Pinctada fucata）中分离到一种具有60%天冬氨酸残基的酸性蛋白Aspein。Aspein的转录本显示对壳的方解石棱柱层部分具有特异性，表明Aspein参与棱柱结构形成和/或方解石沉淀。

项目成果

K.Endo, S.Katsuno, O.Miyoshi, I.Sarashina: "Lymnaea stagnalis as a model organism for studies of calcium carbonate biomineralization"Proceedings of 8^<th> International Symposium on Biomineralization. (未定).

K.Endo、S.Katsuno、O.Miyoshi、I.Sarashina：“Lymnaea stagnalis 作为碳酸钙生物矿化研究的模式生物”第 8 届国际生物矿化研讨会论文集（待定）。

Sarashina, I., Endo, K.: "The complete primary structure of Molluscan Shell Protein 1 (MSP-1), an acidic glycoprotein in the shell matrix of the scallop Patinopecten yessoensis"Marine Biotechnology. 3巻. 362-369 (2001)

Sarashina, I., Endo, K.：“软体动物壳蛋白 1 (MSP-1) 的完整一级结构，这是扇贝虾夷扇贝壳基质中的一种酸性糖蛋白”，海洋生物技术 3. 362-369。 2001）

Sarashina, I. and Endo, K.: "The complete primary structure of Molluscan Shell Protein 1 (MSP-1), an acidic glycoprotein in the shell matrix of the scallop Patinopecten yessoensis"Marine Biotechnology. 3. 362-369 (2001)

Sarashina, I. 和 Endo, K.：“软体动物壳蛋白 1 (MSP-1) 的完整一级结构，软体动物壳蛋白 1 (MSP-1) 是扇贝虾夷扇贝壳基质中的一种酸性糖蛋白”海洋生物技术。

更科功, 小川友美, 遠藤一佳: "軟体動物の殻内酸性糖タンパク質の一次構造"月刊海洋. 32巻. 359-365 (2000)

Isao Sarashina、Tomomi Okawa、Kazuyoshi Endo：“软体动物壳中酸性糖蛋白的一级结构”月刊 Kaiyo Vol. 32. 359-365 (2000)

Endo, K., Sarashina, I., Asami, T.: "Lymnaea stagnalis as a Model Organism for Studies of Calcium Carbonate Biomineralization"Biomineralization : formation, diversity, evolution and application(Kobayashi, I. (ed.))(Tokai Univ. Press.). (未定). (2003)

Endo, K.、Sarashina, I.、Asami, T.：“Lymnaea stagnalis 作为碳酸钙生物矿化研究的模式生物”生物矿化：形成、多样性、进化和应用（小林 I.（编辑））（Tokai （大学出版社）（待定）。