Zalik S E, Thomson L W, Ledsham I M
Department of Zoology, University of Alberta, Edmonton, Canada.
J Cell Sci. 1987 Nov;88 ( Pt 4):483-93. doi: 10.1242/jcs.88.4.483.
The gastrulating chick blastoderm contains lectin activity specific for beta-D-galactoside groups. The galactose-binding lectin isolated by affinity chromatography on rho-aminophenyl-beta-D-lactoside separates into two bands when studied by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. One of these LII has a relative molecular mass of 70 (+/- 2) X 10(3) while the other LI is a polypeptide that migrates with the dye front in 10% gels. We have prepared an antiserum against this lectin preparation and have affinity-purified antibodies against LI. When embryos at stages 3-7 were examined by immunofluorescence using the affinity-purified antibodies, lectin was expressed in cells at the lowest portions of the primitive streak as well as in cells migrating laterally from this region to form the endoderm. Lectin was also expressed by the cells of the extra-embryonic endoderm and the primordial germ cells of the proximal area opaca. In transfers of gradient gels stained with affinity-purified antibodies against LI, this lectin had an approximate molecular weight of 6.5 X 10(3). Our results indicate that this lectin is expressed in areas that are undergoing cell spreading.
原肠胚形成期的鸡胚胚盘含有对β-D-半乳糖苷基团具有特异性的凝集素活性。通过在ρ-氨基苯基-β-D-乳糖苷上进行亲和层析分离得到的半乳糖结合凝集素，经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳研究时可分离成两条带。其中一条LII的相对分子质量为70（±2）×10³，而另一条LI是一种在10%凝胶中与染料前沿一起迁移的多肽。我们制备了针对这种凝集素制剂的抗血清，并对针对LI的抗体进行了亲和纯化。当使用亲和纯化的抗体通过免疫荧光检查3-7期的胚胎时，凝集素在原条最低部分的细胞中表达，也在从该区域横向迁移以形成内胚层的细胞中表达。凝集素也由胚外内胚层细胞和近区暗区的原始生殖细胞表达。在用针对LI的亲和纯化抗体染色的梯度凝胶转移中，这种凝集素的近似分子量为6.5×10³。我们的结果表明，这种凝集素在正在经历细胞铺展的区域表达。
