Extracellular Matrix Macromolecules of Developing Limbs

发育中肢体的细胞外基质大分子

• 批准号: 59570651
• 负责人: OOHIRA Atsuhiko
• 金额: $ 0.96万
• 依托单位: Institute for Developmental Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-59570651/
• 关键词: Limb anomalies; Developing limbs; Rat limb buds; Chondrogenesis; Extracellular matrix; Proteoglycans; Chondroitin sulfate

We have analyzed proteoglycans quantitatively as well as qualitatively in the pathological cartilaginous or osseous tissues excised from patients with various congenital skeletal disorders, especially limb anomalies, and found in most cases abnormalities in the amount and/or structure of proteoglycans. Our observations support the general idea that extracellular matrix macromolecules including proteoglycans are involved in cell differentiation and morphogenesis of skeletal tissues. To clarify the role of proteoglycans in limb morphogenesis, we isolated and characterized proteoglycans synthesized by developing limb buds of the rat.Limb buds, dissected from rat fetuses on day 13, 14 or 15 of gestation, were incubated in a culture medium containing [ <^(35)S> ]sulfate for 3 hr. [ <^(35)S> ]labeled proteoglycans were solubilized in 4 M guanidine HCl containing protease inhibitors and a detergent, 0.8% MEGA-9. A chondroitin sulfate proteoglycan and a heparan sulfate proteoglycan were isolat … More ed from day-13 limb buds. In addition, single chain heparan sulfate was also isolated. Our microscopic examination revealed that mesenchymal-cell condensation to form the primordia of the limb skeleton became observable at this stage. Cartilage-type proteoglycan could be detected for the first time on day 14, and the amount increased thereafter. This finding indicates that chondrogenesis in rat limbs begins on day 14 of gestation.Polysaccharide preparation obtained from day-13 limb buds by proteolysis was separated by two-dimensional electrophoresis on cellulose acetate film into four Alcian blue-stained spots; hyaluronate, heparan sulfate, chondroitin sulfate and an unknown component. Interestingly, the unknown component was not found in polysaccharide preparations obtained either from the brain or tail of rat fetuses at the same developmental stages. Although characterization of this component is not satisfactory at this moment mainly owing to the small amount available, the component could be involved in limb morphogenesis. Less

我们已经分析了蛋白多糖定量以及定性的病理软骨或骨组织切除患者的各种先天性骨骼疾病，特别是肢体畸形，并发现在大多数情况下异常的蛋白多糖的量和/或结构。我们的观察支持了细胞外基质大分子（包括蛋白多糖）参与骨骼组织细胞分化和形态发生的一般观点。为了阐明蛋白聚糖在肢体形态发生中的作用，我们分离并表征了由发育中的大鼠肢芽合成的蛋白聚糖。从妊娠第13、14或15天的大鼠胎儿解剖肢芽，在含有[ <^（35）S> ]硫酸盐的培养基中孵育3小时。将标记的蛋白聚糖溶解在含有蛋白酶抑制剂和去污剂0.8%MEGA-9的4 M盐酸胍中。分离得到硫酸软骨素蛋白聚糖和硫酸乙酰肝素蛋白聚糖， ...更多信息 艾德从第13天的肢芽开始。此外，还分离出单链硫酸乙酰肝素。我们的显微镜检查显示，间充质细胞凝聚形成肢体骨骼的原基成为观察在这个阶段。软骨型蛋白多糖在第14天首次被检测到，此后数量增加。这一发现表明，软骨在大鼠四肢开始在妊娠d14.多糖制备从第13天的肢芽蛋白水解获得的醋酸纤维素薄膜上的双向电泳分离成四个Alcian蓝染色斑点：透明质酸，硫酸乙酰肝素，硫酸软骨素和未知的成分。有趣的是，在从处于相同发育阶段的大鼠胎儿的大脑或尾部获得的多糖制剂中未发现未知组分。虽然目前该组分的表征并不令人满意，主要是由于可用量少，但该组分可能参与肢体形态发生。少

项目成果

Atsuhiko Oohira: Journal of Biochemistry. 99. 1371-1376 (1986)

大平敦彦：生物化学杂志。

野上宏: 整形外科基礎科学. 11. 233-235 (1984)

野上浩：骨科基础科学。11. 233-235 (1984)

Fumiko Matsui: "Glycosaminoglycans and proteoglycans synthesized by rat limb buds during prechondrogenic and chondrogenic stages" Differentiation. 31. 29-34 (1986)

Fumiko Matsui：“大鼠肢芽在软骨形成前和软骨形成阶段合成的糖胺聚糖和蛋白聚糖”分化。

Hiroshi Nogami: "Bone and cartilage of enchondromatosis and metaphysial chondrodysplasia" Orthopedic Research Science. 11. 233-235 (1984)

Hiroshi Nogami：“内生软骨瘤病和干骺端软骨发育不良的骨和软骨”骨科研究科学研究。

Hiroshi Nogami: "Cartilage of mucolipidosis III" Orthopedic Research Science. 13. 1-3 (1986)

Hiroshi Nogami：“粘脂沉积症 III 软骨”骨科研究科学研究。