Research on biological function of ryudocan by knockout mice analysis.

敲除小鼠分析研究龙道康的生物学功能。

• 批准号: 12670981
• 负责人: KOJIMA Tetsuhiro
• 金额: $ 2.43万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670981/
• 关键词: heparan sulfate proteoglvcan; ryudocan; deficient mice; focal adhesion; fibronectin; renal damage; K-carrageenan; obstructive nephropathy; カッパーカラゲナン; マウス; 細胞接着

Two domains of fibronectin deliver two different but cooperative signals required for focal adhesion formation. The signal from the cell-binding domain is mediated by integrins, whereas the signal from the heparin binding domain is recognized by heparan sulfate proteoglycans, of which syndecan-4 (ryudocan) has been hypothesized to be involved in focal adhesion formation. We generated mice deficient in syndecan-4 to study its role directly. Even in fibroblasts from svndecan-4-deficient mice, focal adhesions were formed, and actin fibers terminated normally at focal adhesions when they were cultured on coverslips coated with fibronectin or with a mixture of its cell-binding and heparin binding fragments. However, when the cells were cultured on the cell-binding fragment and the heparin-binding fragment was added to the medium, focal adhesion formation was impaired in the syndecan-4 null fibroblasts as compared with that in wild-type cells. Therefore, syndecan-4 is essential for promoting … More focal adhesion formation only when the signal of the heparin-binding domain of fibronectin is delivered as a soluble form, most probably from the apical surface. When the signal is delivered as a substratum-bound form, other molecule(s) also participate(s) in the signal reception.The expression and roles of syndecan-4 in the kidney were investigated. Syndecan-4 expression was detected in the ureteric bud invaginating into the metanephric mesenchyme at 11.5 gestational days, and remained in the collecting ducts, distal renal tubules, glomeruli and some capillaries between renal tubules until the mature kidney stage. However, organogenesis of the kidney was normal in syndecan-4-deficient [Synd4(-/-)] mice. Although most of renal functions of Synd4(-/-) mice were not impaired,significant increase of susceptibility to the k-carrageenan-induced renal damage was observed in Synd4(-/-) mice. K-Carrageenan was deposited severely in the collecting ducts of Synd4(-/-) mice and caused obstructive nephropathy. Leading to death of 7 of 24 Synd4(-/-) mice within 7 days after administration, whereas none of 24 Synd4(+/+) mice died. After the administration ofk carrageenan, blood urea nitrogen of Syrid4(-/-) mice was significantly higher than that of Synd4(+/+) mice, k-Carrageenan had affinity to the membrane fraction derived from the medulla of the kidney, and its binding was inhibited by heparin. Thus, syndecan-4 may function to prevent k-carrageenan deposition in the collecting ducts via its heparan sulfates. Less

纤连蛋白的两个结构域传递两种不同但协同的粘着斑形成所需的信号。来自细胞结合结构域的信号由整联蛋白介导，而来自肝素结合结构域的信号由硫酸乙酰肝素蛋白聚糖识别，其中多配体聚糖-4（ryudocan）已被假设参与粘着斑形成。我们产生了syndecan-4缺陷的小鼠，以直接研究其作用。即使在svndecan-4缺陷小鼠的成纤维细胞中，也形成了局灶性粘连，当它们在用纤连蛋白或其细胞结合和肝素结合片段的混合物包被的盖玻片上培养时，肌动蛋白纤维通常终止于局灶性粘连。然而，当在细胞结合片段上培养细胞并将肝素结合片段加入培养基中时，与野生型细胞相比，syndecan-4无效成纤维细胞中的粘着斑形成受损。因此，syndecan-4对于促进 ...更多信息 只有当纤连蛋白的肝素结合结构域的信号以可溶形式传递时，最可能是从顶端表面传递时，才形成粘着斑。当信号以基质结合的形式传递时，其他分子也参与信号的接收。Syndecan-4在妊娠11.5 d时表达于输尿管芽内陷至后肾间充质中，并持续存在于集合管、远端肾小管、肾小球和肾小管间的毛细血管中，直至肾成熟期。然而，在syndecan-4-缺陷[Synd 4（-/-）]小鼠中，肾脏的器官形成是正常的。虽然Synd 4（-/-）小鼠的大部分肾功能未受损，但在Synd 4（-/-）小鼠中观察到对k-角叉菜胶诱导的肾损伤的易感性显著增加。角叉菜胶在Synd 4（-/-）小鼠的集合管中沉积严重，并引起阻塞性肾病。导致24只Synd 4（-/-）小鼠中有7只在给药后7天内死亡，而24只Synd 4（+/+）小鼠无一死亡。角叉菜胶可使Syrid 4（-/-）小鼠的血尿素氮显著高于Synd 4（+/+）小鼠，角叉菜胶与肾髓质膜组分具有亲和性，肝素可抑制其结合。因此，多配体聚糖-4可以通过其硫酸乙酰肝素起作用以防止k-角叉菜胶沉积在集合管中。少

项目成果

T. Shimokawa: "Expression of Protein S in the Murine Heart and Cultured Mouse Cardiomyocytes, Is Down-regulated by Cytokines"Thromb. Haemost.. 86. 623-629 (2001)

T. Shimokawa：“小鼠心脏和培养的小鼠心肌细胞中蛋白质 S 的表达受到细胞因子的下调”Thromb。

K.Iba,R.Albrechtsen,T.Kojima., et al.: "Factor X Nagoya 1 and 2 : A CRM-Factor X deficiency and A Dysfunctional CRM+ Factor X Deficiency Characterized by Substitution of Arg306 by Cys and Gly366 by Ser."J.Cell.Biol.. 149(5). 1143-1155 (2000)

K.Iba、R.Albrechtsen、T.Kojima. 等人：“因子 X Nagoya 1 和 2：CRM-因子 X 缺乏症和功能失调的 CRM 因子 X 缺乏症，其特征是 Arg306 被 Cys 取代，Gly366 被 Ser 取代。

K. Ishiguro: "Syndecan-4 Deficiency Increase Susceptability to k-Carrageenan-Induced Renal Damage."Lab. Invest.. 81. 509-516 (2001)

K. Ishiguro：“Syndecan-4 缺乏会增加对 k-卡拉胶引起的肾损伤的敏感性。”实验室。

小嶋哲人: "新しいDICの病態・診断・治療 III.治療 4.抗凝固薬 1)ヘパリン 2)低分子ヘパリン 3)ヘパラン硫酸"医薬ジャーナル社. 6 (2001)

小岛哲人：“DIC的新病理学、诊断和治疗III.治疗4.抗凝剂1)肝素2)低分子肝素3)硫酸乙酰肝素”医药杂志社6(2001)

M.Sakata,H.Kobayashi,T.Kojima, et al.: "Ryudocan expression by lueinized granulosa cells is associated with the process of follicle atresia.."Fertll.Sterll.. 74(6). 1208-1214 (2000)

M.Sakata、H.Kobayashi、T.Kojima 等人：“黄素化颗粒细胞表达的 Ryudocan 与卵泡闭锁过程相关。”Fertll.Sterll.. 74(6)。