Notochordal stimulation of in vitro somite chondrogenesis before and after enzymatic removal of perinotochordal materials.
Notochordal stimulation of in vitro somite chondrogenesis before and after enzymatic removal of perinotochordal materials.
复制标题
酶促去除脊索周围材料之前和之后对体外体节软骨形成的脊索刺激。
DOI：
10.1016/0012-1606(75)90340-1
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发表时间：
1975
影响因子：
2.7
通讯作者：
J. Lash
中科院分区：
文献类型：
--
作者：
R. Kosher;J. Lash
在目前的研究中，有证据直接表明胚胎脊索产生的蛋白多糖在控制体软骨形成中起作用。几种组织化学技术已经证明，在与some相互作用期间，脊索合成了脊索周围蛋白聚糖，这些蛋白聚糖已被证明含有4-硫酸软骨素（40%），6-硫酸软骨素（40%）和硫酸肝素（20%）。在胰蛋白酶的帮助下从胚胎中剥离脊索会导致脊索周围细胞外基质物质的去除，包括蛋白聚糖，而在没有酶处理或低浓度胶原酶的帮助下剥离脊索会导致它们的保留。软骨形成的速度和数量有相当大的增加，在保留脊索周围物质的条件下，与脊索解剖相关的某些体所积累的硫酸糖胺聚糖的数量相应增加2至3倍。用高度纯化的酶（软骨素酶ABC、AC和睾丸透明质酸酶）治疗胶原酶解剖或自由解剖的脊索，这些酶能特异性地降解蛋白聚糖，导致组织化学可检测到的周围脊索蛋白聚糖的丢失。这些脊索支持壳体软骨形成的能力严重受损。此外，当胰蛋白酶处理的脊索在营养琼脂（没有体体）上培养（“预培养”）24小时时，脊索周围物质重新积累。与未经“预培养”的胰蛋白酶处理的脊索相比，与这种“预培养”的脊索一起培养的索体在形成的软骨数量上有相当大的增加，积累的硫酸糖胺聚糖的数量增加了2- 3倍。这一观察结果表明，经胰蛋白酶处理的脊索通过重新合成和积累脊索周围物质，重新获得了最大限度地刺激镜质体软骨形成的能力。最后，“预培养”的脊索用软骨素酶去除脊索周围的蛋白聚糖，其支持软体软骨形成的能力明显受损。这些观察结果与胚胎脊索产生的蛋白聚糖在体软骨形成中起重要作用的概念是一致的。
In the present investigation, evidence is presented directly implicating proteoglycans produced by the embryonic notochord in the control of somite chondrogenesis. It has been demonstrated by several histochemical techniques that during the period of its interaction with somites, the notochord synthesizes perinotochordal proteoglycans, and these proteoglycans have been shown to contain chondroitin 4-sulfate (40%), chondroitin 6-sulfate (40%), and heparan sulfate (20%). Dissection of notochords from embryos with the aid of a brief treatment with trypsin results in the removal of perinotochordal extracellular matrix materials including proteoglycans, while dissection of notochords without the aid of enzyme treatment or with a low concentration of collagenase results in their retention. There is a considerable increase in the rate and amount of cartilage formation and a corresponding 2 to 3-fold increase in the amount of sulfated glycosaminoglycan accumulated by somites cultured in association with notochords dissected under conditions in which perinotochordal materials are retained. Treatment of collagenase-dissected or freely dissected notochords with highly purified enzymes (chondroitinase ABC, AC, and testicular hyaluronidase) which specifically degrade proteoglycans causes a loss of histochemically detectable perinotochordal proteoglycans. These notochords are considerably impaired in their ability to supportin vitrosomite chondrogenesis. In addition, when trypsin-treated notochords are cultured (“precultured”) for 24 hr on nutrient agar (in the absence of somites), perinotochordal material reaccumulates. Somites cultured in association with such “precultured” notochords exhibit considerable increase in the amount of cartilage formed and a 2- to 3-fold increase in the amount of sulfated glycosaminoglycan accumulated as compared to somites cultured in association with trypsin-treated notochords which have not been “precultured.” This observation indicates that trypsin-treated notochords reacquire their ability to maximally stimulatein vitrosomite chondrogenesis by resynthesizing and accumulating perinotochordal material. Finally, “precultured” notochords treated with chondroitinase to remove perinotochordal proteoglycans are considerably impaired in their ability to supportin vitrosomite chondrogenesis. These observations are consonant with the concept that proteoglycans produced by the embryonic notochord play an important role in somite chondrogenesis.