Hurle J M, Ros M A, Hinchliffe J R
Departamento de Anatomia y Biologia Celular, Facultad de Medicina, Universidad de Cantabria, Spain.
Cell Differ. 1988 Jul;24(2):149-58. doi: 10.1016/0045-6039(88)90066-8.
The changing pattern of expression of glycoconjugates during the differentiation of the chick leg bud between stages 17 to 34 (days 3 to 8 of incubation) was studied using fluorochrome-labelled plant lectins. Limb buds were fixed in cold acetic-alcohol and wax-embedded. Agglutinins of peanut (PNA), soybean (SBA) and succinylated wheat germ (WGAs) revealed a specific binding pattern in the apical ectodermal ridge (AER) between Hamburger and Hamilton stages 19-32. These stages coincide with the period of elevation of the AER. This specific binding pattern was absent from the adjacent dorsal and ventral ectoderm. Prechondrogenic cells were positive for WGA and for PNA, and the PNA-binding capacity was intensified after neuraminidase treatment. Premyogenic cells at stage 23 can be identified as negative to PNA after neuraminidase, while the blood vessels became positive. PNA, SBA, WGA, WGAs and, in addition, Ricinus communis (RCA-I) lectins stained the basal membrane. Strands of extracellular matrix which connect with the basal membrane and cross the limb transversely between dorsal and ventral ectoderm were stained by RCA-I, SBA and PNA after neuraminidase.
利用荧光染料标记的植物凝集素，研究了鸡胚腿芽在第17至34阶段（孵化第3至8天）分化过程中糖缀合物表达模式的变化。肢芽用冷醋酸酒精固定并包埋于石蜡中。花生凝集素（PNA）、大豆凝集素（SBA）和琥珀酰化麦胚凝集素（WGA）在汉伯格和汉密尔顿第19 - 32阶段的顶端外胚层嵴（AER）中显示出特定的结合模式。这些阶段与AER升高的时期一致。相邻的背侧和腹侧外胚层没有这种特定的结合模式。软骨前体细胞对WGA和PNA呈阳性，经神经氨酸酶处理后PNA结合能力增强。第23阶段的前成肌细胞经神经氨酸酶处理后对PNA呈阴性，而血管呈阳性。PNA、SBA、WGA、琥珀酰化麦胚凝集素以及蓖麻凝集素（RCA-I）对基底膜进行染色。经神经氨酸酶处理后，与基底膜相连并在背侧和腹侧外胚层之间横向穿过肢体的细胞外基质条带被RCA-I、SBA和PNA染色。
