Signal molecules in the jelly coat of starfish eggs for the inducing of acrosome reaction.

海星卵果冻层中用于诱导顶体反应的信号分子。

• 批准号: 08458237
• 负责人: HOSHI Motonori
• 金额: $ 5.44万
• 依托单位: Tokyo Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08458237/
• 关键词: acrosome reaction; sperm-egg binding; starfish; キヒトデ; 糖鎖

The acrosome reaction (AR) is a species-specific exocytosis response of spermatozoa to the egg envelopment. In the starfish Asterias amurensis, three components in the jelly coat of eggs act in concert on spermatozoa to elicit the AR ; namely acrosome reaction-inducing substance (ARIS), Co-ARISs and sperm-activating peptides named asterosaps. ARIS is highly sulfated glycoprotein of an apparent molecular size over 10^7 and its activity is mostly attributable to sulfated sugar chains. The main saccharide chain of ARIS is a very long (2 x 10^6 in more in molecular size) linear polymer of the pentasaccharide repeating units (Fr.1) is able to induce the AR at high calcium concentration as well as ARIS does. To obtain binding parameters of an ARIS receptor, binding experiments were carried out by centrifugation binding assay using radio-labeled Fr.1. Spermatozoa were incubated with radioactive iodinated Fr.1. With Scatchard plot, there are two type receptor, High affinity type (Kd ; 145 muM,Bmax ; 8.38 x 10^4) and low affinity type (Kd ; 4.43 x 10^2muM,Bmax ; 1.89 x 10^6). And competitive assay were carried with non-labeled Fr.1 and ^<125>I labeled Fr.1. The bound and free concentration of the ligand were measured. The result showed that non-labeled Fr.1 inhibits to bind labeled Fr.1.

Adrosom反应（AR）是精子对卵包膜的物种特异性胞吐反应。在海星阿美素（Amurensis）中，果冻外套中的三个成分在精子上演奏，以引起AR。即诱导的Adrosom反应物质（ARIS），称为星形胶囊的同时和精子激活的肽。 ARIS是高度硫化的糖蛋白，其在10^7上的明显分子大小，其活性主要归因于硫酸化的糖链。 ARIS的主要糖链是五糖重复单元（FR.1）的线性聚合物非常长的（2 x 10^6）能够像ARIS一样在高钙浓度下诱导AR。为了获得ARIS受体的结合参数，使用射线标记的FR.1通过离心结合测定进行结合实验。精子与放射性碘化的FR.1一起孵育。使用Scatchard图，有两种类型的受体，高亲和力类型（KD； 145 MUM，BMAX； 8.38 x 10^4）和低亲和力类型（KD; 4.43 x 10^2^2mum，bmax; 1.89 x 10^6）。并用未标记的FR.1和 ^<125>标记为Fr.1的竞争测定法。测量配体的结合和游离浓度。结果表明，未标记的FR.1抑制结合标记的FR.1。

项目成果

M.Ogiso, M.Komoto and M.Hoshi: "Identification and Synthetic Pathway of Sialyl-Lewis^x-containing Neolacto-series Gangliosides in Lens Tissues. 2. Enzymatic Synthesis of Sialyl-Lewis^x Gangliosides in Monkey and Rat Lenses" Biochim.Biophys.Acta.1315. 29-3

M.Ogiso、M.Komoto 和 M.Hoshi：“晶状体组织中含有 Sialyl-Lewis^x 的 Neolacto-series 神经节苷脂的鉴定和合成途径。2. 猴子和大鼠晶状体中 Sialyl-Lewis^x 神经节苷脂的酶法合成”

M.Matsumoto, F.Saito-Ohara, M.Hoshi and S.Kurata: "Cloning and Sequencing of Mouse Complementary DNA for Heme Oxygenase-2" J.Biochem.120. 1079-1081 (1996)

M.Matsumoto、F.Saito-Ohara、M.Hoshi 和 S.Kurata：“血红素加氧酶 2 的小鼠互补 DNA 的克隆和测序”J.Biochem.120。

T.Nishigaki, K.Chiba and M.Hoshi: "Structure and function of asterosaps,sperm-activating peptides from the jelly coat of starfish eggs." J.Rep.Dev.43 Suppl. 115-116 (1997)

T.Nishigaki、K.Chiba 和 M.Hoshi：“星形囊的结构和功能，来自海星卵果冻层的精子激活肽。”

K.Chiba, E.Sato, and M.Hoshi: "Detection of In Vivo Proteasome Activity in a Starfish Oocyte Using Membrane-Impermeant Substrate" J.Biochem. 122. 286-293 (1997)

K.Chiba、E.Sato 和 M.Hoshi：“使用非膜性基质检测海星卵母细胞体内蛋白酶体活性”J.Biochem。

M.Byrne, A.Cerra, T.Nishigaki and M.Hoshi: "Infestation of the testes of the Japanese sea star Asterias amurensis by the ciliate Orchitophyra stellarum : a caution against the use of this ciliate for biological control" Dis.Aquat.Org.28. 235-239 (1997)

M.Byrne、A.Cerra、T.Nishigaki 和 M.Hoshi：“日本海星 Asterias amurensis 的睾丸被纤毛虫 Orchitophyra stellarum 侵染：警告不要使用这种纤毛虫进行生物防治”Dis.Aquat。