Roles of cell adhosive peptides from laminins in organogenesis

层粘连蛋白细胞粘附肽在器官发生中的作用

基本信息

批准号：
10670028
负责人：
KADOYA Yuichi
金额：
$ 1.73万
依托单位：
KITASATO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

项目摘要

1. The distributions of laminin-5 and laminin-α5 chain, were studied immunohistochemically in the developing submandibular glands of mice. The expression of laminin-5 was found in the basement membrane of developing duct but not epithelial end bud. In contrast, laminin-α5 chain was expressed in the entire epithelial basement membrane at the early embryonic stage, suggesting the role of laminin-α5 chain on the epithelial morphogenesis.2. To elucidate roles of the laminin α5 chain on organogenesis, we tested 15 cell-adhesive synthetic peptides from a carboxy-terminal globular domain of laminin-α5-chain sequence in submandibular gland organ culture. We found a peptide sequence LVLFLNHGHFVA (residues 3307-3318 and designated A5G-77) strongly inhibits the organ development at 250μg/ml. Various amino- and carboxy-terminal truncated peptides of A5G-77 were synthesized and their activities were also tested in the same system. A carboxy-treminal deletion peptide A5G-77f (LVLFLNHGH) worked inhib … More itory, whereas an amino-treminal deletion peptide A5G-77c (FLNHGHFVA) failed to inhibit the organ development. The morphology of A5G-77-treated explants differed from those of AG-73-treated, indicating distinct role of individual laminin α chains for organ development.3. We established a serum-free organ culture system of isolated vibrissal rudiments of embryonic day-13 mice. This system allowed us to test 22 laminin-1 peptides for their roles on the vibrissal growth and differentiation. We found that RKRLQVQLSIRT sequence, which is mapped at LG-4 subdomain of laminin-α1-chain carboxyl-terminal G-domain and designated AG-73, perturbed vibrissal growth in vitro. Other cell-adhesive laminin-1 peptides and a control scrambled-peptide, LQQRRSVLRTKI, failed to show any significant effects on vibrissal growth. An impaired basement membrane formation, an abnormal cytoplasmic bleb formation, and an unusual basal formation of actin bundles were noted in AG-73-treated vibrissal epidermis, indicating that AG-73 signaling is involved in cytoskeletal organization. Less

1.免疫组化研究小鼠颌下腺发育过程中层粘连蛋白5和层粘连蛋白α5链的分布。在发育中的下颌下腺中发现层粘连蛋白5的表达，但在上皮终芽中未发现层粘连蛋白5的表达。 -α5链在胚胎早期在整个上皮基底膜中表达，提示层粘连蛋白-α5链对上皮细胞的作用2.为了阐明层粘连蛋白α5链在器官发生中的作用，我们在颌下腺器官培养物中测试了来自层粘连蛋白-α5链序列的羧基末端球状结构域的15种细胞粘附合成肽。残基 3307-3318 并命名为 A5G-77) 强烈抑制器官发育合成了 250μg/ml 的 A5G-77 的各种氨基末端和羧基末端截短肽，并在同一系统中测试了它们的活性。羧基末端缺失肽 A5G-77f (LVLFLNHGH) 具有抑制作用，而氨基末端缺失肽A5G-77c (FLNHGHFVA)未能抑制器官发育。 A5G-77处理的外植体与AG-73处理的外植体不同，表明单个层粘连蛋白α链对于器官发育的不同作用。3．我们建立了胚胎第13天小鼠的分离触毛雏形的无血清器官培养系统。该系统使我们能够测试 22 个层粘连蛋白-1 肽在触须生长和分化中的作用，我们发现 RKRLQVQLSIRT 序列，其映射于层粘连蛋白 α1 链羧基末端 G 结构域的 LG-4 子结构域和指定的 AG-73，在体外干扰了触毛生长。其他细胞粘附层粘连蛋白 1 肽和对照乱序肽 LQQRRSVLRTKI 未能显示任何显着的结果。基底膜形成受损、细胞质泡形成异常以及肌动蛋白束基底形成异常。在 AG-73 处理的触须表皮中注意到，表明 AG-73 信号传导参与细胞骨架组织。