Zschäbitz A, Weiser H, Stofft E, Krahn V, Gabius H J, Khaw A, Biesalski H K
Anatomisches Institut der Johannes Gutenberg-Universität Mainz, Germany.
Anat Embryol (Berl). 1995 Jan;191(1):47-9. doi: 10.1007/BF00215297.
The staining patterns of 24 biotinylated lectins were analyzed in serial sections of the mandible of 13- to 21-day-old rat embryos by means of the avidin-biotin-peroxidase method. A ubiquitous distribution of binding sites was demonstrated after incubation with Con A (Canavalia ensiformis), DSL (Datura stramonium; except bone matrix), and WGA (Triticum vulgare). ECL (Erythrina cristagalli), GSL I (Griffonia simplicifolia), SJA (Saphora japonica), VVL (Vicia villosa), DBA (Dolichus biflorus), UEA I (Ulex europeus), and LTA (Lotus tetragonobolus) were constantly negative. In early stages of development, GSL II (Griffonia simplicifolia II) was a selective marker of prechondral blastema. In contrast, PNA (Arachis hypogaea) did not stain condensing mesenchyme. During chondrogenesis of Meckels's cartilage a general decrease of lectin binding was observed. Mature cartilage matrix was constantly negative. Chondrocytes were marked by the lectins PSA (Pisum sativum), WGA, PHA-E, and PHA-L (Phaseolus vulgaris E and L). A strong GSL II binding was restricted to the mesial-superior region of the perichondrium. In later stages, several lectins revealed significant differences between preskeletal ("central") areas and the remaining ("peripheral") mesenchyme. A clear binding reaction was noted in central regions by applying LEA (Lycopersicon esculentum) and STL (Solanum tuberosum), while the peripheral tissue was only faintly stained. Developing bone was specifically marked by succinylated WGA (sWGA). The lectins LCA (Lens culinaris) and RCA (Ricinus communis) bound to fibers and extracellular matrix of the connective tissue. Jacalin (Artocarpus integrifolia) and SBA (Glycine max) binding sites were found in macrophages. Affinity of VAA (Viscum album) increased parallel with maturation of endothelial cells. Specific lectin-binding patterns revealed no correlation with the distribution of glycosaminoglycans. The results demonstrate a general reduction of oligosaccharide structures during development of Meckel's cartilage. From our observations we conclude that intralaminar glucose and/or mannose sequences as well as terminal sialic acid molecules are ubiquitously distributed, while terminal alpha-fucose was constantly negative. Lectin-binding patterns of macrophages may reflect the presence of specifically linked terminal galactose. Our findings indicate that oligosaccharides terminating in N-acetylglucosamine are bone-specific. The significance of the restricted staining of the perichondrium by GSL II remains to be elucidated.
采用抗生物素蛋白-生物素-过氧化物酶法，对13至21日龄大鼠胚胎下颌骨连续切片中24种生物素化凝集素的染色模式进行了分析。用刀豆球蛋白A（刀豆）、DSL（曼陀罗；骨基质除外）和麦胚凝集素（普通小麦）孵育后，显示出结合位点的普遍分布。鸡冠花凝集素（刺桐）、GSL I（西非单叶豆）、槐凝集素（槐树）、绒毛豆凝集素（绒毛豆）、双花扁豆凝集素（双花扁豆）、荆豆凝集素I（荆豆）和四角豆凝集素（四角豆）始终呈阴性。在发育早期，GSL II（西非单叶豆II）是软骨前胚基的选择性标记。相比之下，花生凝集素（花生）不染色凝聚间充质。在梅克尔软骨的软骨形成过程中，观察到凝集素结合普遍减少。成熟软骨基质始终呈阴性。软骨细胞以凝集素PSA（豌豆）、麦胚凝集素、PHA-E和PHA-L（菜豆E和L）标记。强烈的GSL II结合仅限于软骨膜的内侧上部区域。在后期，几种凝集素显示出骨骼前（“中央”）区域和其余（“周边”）间充质之间的显著差异。通过应用番茄凝集素（番茄）和马铃薯凝集素（马铃薯）在中央区域观察到明显的结合反应，而周边组织仅轻微染色。发育中的骨骼以琥珀酰化麦胚凝集素（sWGA）特异性标记。凝集素LCA（小扁豆）和蓖麻凝集素（蓖麻）与结缔组织的纤维和细胞外基质结合。在巨噬细胞中发现了木菠萝凝集素（面包果）和大豆凝集素（大豆）的结合位点。槲寄生凝集素（槲寄生）的亲和力随着内皮细胞的成熟而平行增加。特异性凝集素结合模式与糖胺聚糖的分布无关。结果表明，在梅克尔软骨发育过程中寡糖结构普遍减少。根据我们的观察，我们得出结论，层内葡萄糖和/或甘露糖序列以及末端唾液酸分子普遍分布，而末端α-岩藻糖始终呈阴性。巨噬细胞的凝集素结合模式可能反映了特异性连接的末端半乳糖的存在。我们的研究结果表明，以N-乙酰葡糖胺结尾的寡糖是骨骼特异性的。GSL II对软骨膜的局限性染色的意义仍有待阐明。
