Sherratt M J, Holmes D F, Shuttleworth C A, Kielty C M
School of Biological Sciences, University of Manchester, U.K.
Int J Biochem Cell Biol. 1997 Aug-Sep;29(8-9):1063-70. doi: 10.1016/s1357-2725(97)00028-9.
We have applied scanning transmission electron microscopy to intact native fibrillin-containing microfibrils isolated from foetal bovine elastic tissues in order to derive new insights into microfibril organisation. This technique provides quantitative data on the mass per unit length and axial mass distribution of unstained, unshadowed macromolecules. Scanning transmission electron microscopy of microfibrils from aorta, skin and nuchal ligament revealed that the beads corresponded to peaks of mass and the interbead regions to troughs of mass. These major features of axial mass distribution were characteristic of all microfibrils examined. Tissue-specific and age-dependent variations in mass were identified in microfibrils that were structurally comparable by rotary shadowing electron microscopy. Increased microfibril mass correlated with increasing gestational age. The additional mass was associated predominantly at, or close to, the bead. Some microfibril populations exhibited pronounced assymetry in their axial mass distribution. These data indicate that intact native microfibrillar assemblies from developing elastic tissues are heterogeneous in composition. Loss of mass following chondroitinase ABC or AC lyase treatment confirmed the presence of chondroitin sulphate in nuchal ligament microfibrillar assemblies.
我们已将扫描透射电子显微镜应用于从胎牛弹性组织中分离出的完整天然含原纤维蛋白微原纤维,以便对微原纤维的组织结构有新的认识。该技术提供了关于未染色、未投影大分子的单位长度质量和轴向质量分布的定量数据。对来自主动脉、皮肤和项韧带的微原纤维进行扫描透射电子显微镜观察发现,珠状物对应于质量峰值,珠间区域对应于质量谷值。轴向质量分布的这些主要特征是所有检查过的微原纤维的特征。通过旋转投影电子显微镜在结构上具有可比性的微原纤维中发现了质量的组织特异性和年龄依赖性变化。微原纤维质量的增加与胎龄的增加相关。额外的质量主要与珠状物处或其附近相关。一些微原纤维群体在其轴向质量分布中表现出明显的不对称性。这些数据表明,来自发育中的弹性组织的完整天然微原纤维组件在组成上是异质的。用软骨素酶ABC或AC裂解酶处理后质量的损失证实了项韧带微原纤维组件中存在硫酸软骨素。