The synthesis and localization of glycosaminoglycans in striated muscle differentiating in cell culture.
The synthesis and localization of glycosaminoglycans in striated muscle differentiating in cell culture.
复制标题
细胞培养中分化的横纹肌中糖胺聚糖的合成和定位。
DOI：
--
复制
发表时间：
1977
期刊：
影响因子：
--
通讯作者：
S. Meier
中科院分区：
文献类型：
--
作者：
P. Ahrens;M. Solursh;S. Meier
第3天大量肌肉培养物将35 SO 4 =和3 H·葡糖胺掺入到通过多种方法（酶敏感性、亚硝酸降解、电泳、薄层色谱法）鉴定的特定糖胺聚糖中。超过75%的新合成的糖胺聚糖出现在培养基中。在培养基中，糖胺聚糖主要由透明质酸（约35%）和硫酸软骨素（约40%）组成，含有少量的软骨素（约15%）和硫酸乙酰肝素（约10%）。在保留在细胞层中的糖胺聚糖中，硫酸软骨素（约45%）和硫酸乙酰肝素（20-40%）占主导地位，仅存在少量的软骨素（约15%）和透明质酸（约15%）。在培养基和细胞层中，大于80%的硫酸软骨素是6-硫酸软骨素。尽管大量培养物中含有一些非肌细胞类型，这无疑有助于总糖胺聚糖的合成，但与肌肉克隆细胞相关的放射自显影颗粒对睾丸透明质酸酶消化的敏感性表明，至少有一些糖胺聚糖（特别是硫酸软骨素）是由肌细胞合成的。
透射电子显微镜观察发现，肌管表面沿着存在钌红染色的球状单位。链霉菌透明质酸酶和睾丸透明质酸酶破坏表面相关钌红染色材料的完整性表明，透明质酸和硫酸软骨素都存在于肌肉细胞表面。
Day 3 mass muscle cultures incorporated 35SO4= and 3H· glucosamine into specific glycosaminoglycans that were identified by a variety of methods (enzyme susceptibility, nitrous acid degradation, electrophoresis, thin layer chromatography). More than 75% of the newly synthesized glycosaminoglycans appeared in the medium. In the medium, the glycosaminoglycans consisted of primarily hyaluronic acid (∼35%) and chondroitin sulfate (∼40%) with smaller amounts of chondroitin (∼15%) and heparan sulfate (∼10%). Among the glycosaminoglycans that remained in the cell layer, chondroitin sulfate (∼45%) and heparan sulfate (20-40%) predominated and only a small amount of chondroitin (∼15%) and hyaluronic acid (∼5%) were present. In both the medium and cell layer, greater than 80% of the chondroitin sulfate was chondroitin-6-sulfate. Although mass cultures contain some non-muscle cell types which undoubtedly contributed to total glycosaminoglycan synthesis, susceptibility of the autoradiographic grains associated with cells in muscle clones to testicular hyaluronidase digestion suggests that at least some glycosaminoglycans (chondroitin sulfate, in particular) were synthesized by muscle cells.
Transmission electron microscopy revealed the presence of ruthenium red staining globular units along the surfaces of myotubes. Destruction of the integrity of surface associated ruthenium red stained material by both Streptomyces hyaluronidase and testicular hyaluronidase suggests that both hyaluronic acid and chondroitin sulfate are present on muscle cell surfaces.