Stoddart R W, Jones C J
Department of Pathological Sciences, Stopford Building, University of Manchester, UK.
Methods Mol Med. 1998;9:21-39. doi: 10.1385/0-89603-396-1:21.
Within the last 25 yr, the study of glycans, their structures, and their drstribution in tissues has emerged from relative obscurity to become a major theme of molecular and cellular biology: "glycobiology" (1). Glycans are major components of cellular surfaces (2,3), extracellular matrices (4,5), and secretions (6), and play important roles in cell-cell and cell-matrix recognition and adhesion (7,8). They regulate the surface environment of cells by influencing the structure of water (6,9), by modulating diffusion, by sequestering metabohtes such as metal ions, by presenting various growth factors to their receptors (10,11), by acting as ligands in recognition-adhesion systems (11-13), and by making major contributions to cell surface charge (2,14). In secretions, they are variously determinants of molecular folding (15), hydration, interaction, and targeting, and they serve in the mechanisms that monitor the aging of glycoconjugates in circulation (6,9,16,17). They are now implicated in mechanisms of molecular targeting and segregation within the endomembrane systems of cells (18-20), in calcium transport in mitochondria (21), the handling of mRNA and its export from the nucleus, the regulation of transcription (19), and in then classical role as energy stores. In all of this, the association of anatomical or ultrastructural localization with biological function is of paramount importance. specific glycans occur in specific places (22,23). The challenge for the histochemist is to reconcile the achievement of sufficiently precise anatomical localization of glycans with the maximum of chemical information about their nature. The lectins have been the main means of accompllshing this, since their first application as fluorochrome-labeled probes to paraffin sections in the early 1970s (2,24). More recently, fluorescence has been largely supplanted by nonfluorescent disclosing systems, such as biotin-avidin-peroxidase and, though paraffin sections are still widely used, resin-embedding of specimens has proved advantageous in terms of resolution and economy of material at the light microscopic level.
在过去的25年里,聚糖及其结构以及它们在组织中的分布的研究已从相对默默无闻发展成为分子和细胞生物学的一个主要主题:“糖生物学”(1)。聚糖是细胞表面(2,3)、细胞外基质(4,5)和分泌物(6)的主要成分,在细胞间和细胞与基质的识别及黏附中发挥重要作用(7,8)。它们通过影响水的结构(6,9)、调节扩散、螯合金属离子等代谢物、向其受体呈递各种生长因子(10,11)、在识别-黏附系统中充当配体(11 - 13)以及对细胞表面电荷做出主要贡献(2,14)来调节细胞的表面环境。在分泌物中,它们在分子折叠(15)、水合作用、相互作用和靶向作用方面起着不同的决定作用,并且它们参与监测循环中糖缀合物老化的机制(6,9,16,17)。它们现在涉及细胞内膜系统内的分子靶向和分离机制(18 - 20)、线粒体中的钙运输(21)、mRNA的处理及其从细胞核的输出、转录的调节(19)以及它们作为能量储存的经典作用。在所有这些方面,解剖学或超微结构定位与生物学功能的关联至关重要。特定的聚糖出现在特定的位置(22,23)。组织化学家面临的挑战是在实现聚糖足够精确的解剖学定位的同时,获取关于其性质的最大化学信息。自20世纪70年代初首次作为荧光染料标记探针应用于石蜡切片以来(2,24),凝集素一直是实现这一目标的主要手段。最近,荧光在很大程度上已被非荧光显色系统所取代,如生物素 - 抗生物素蛋白 - 过氧化物酶,并且尽管石蜡切片仍被广泛使用,但在光学显微镜水平上,标本的树脂包埋在分辨率和材料经济性方面已被证明具有优势。