Blanks J C, Johnson L V
J Comp Neurol. 1983 Nov 20;221(1):31-41. doi: 10.1002/cne.902210103.
A battery of eight lectins with different carbohydrate specificities was used to study changes in glycoconjugate expression during cell differentiation in the mouse retina. The lectins tested included concanavalin A (Con A), wheat germ agglutinin (WGA), soybean agglutinin (SBA), peanut agglutinin (PNA), Ulex europaeus agglutinin (UEA), Ricinus communis agglutinin I (RCA), Dolichos biflorus agglutinin (DBA), and Limulus polyphemus agglutinin (LPA). Unfixed frozen sections of adult and early postnatal mouse retina were treated with fluorescein isothiocyanate-conjugated lectins and examined by fluorescence microscopy. The results showed selective lectin binding in both cellular and synaptic retinal layers of the adult mouse and throughout postnatal development. In general, an increase in intensity of fluorescent lectin staining during retinal development was observed for Con A, WGA, DBA, LPA, RCA, and PNA. This suggests an increase in the expression or accessibility of carbohydrate moieties during development. SBA and UEA showed little to no binding to adult or neonatal retina. Retinal vasculature was intensely stained by RCA, both during development and in the adult. All lectins binding to adult or neonatal retinal layers showed some degree of reactivity with the inner segment region of photoreceptor cells. However, only Con A, PNA and WGA bound to photoreceptor outer segments, suggesting significant differences in the glycosylated components of inner and outer segment membranes. PNA bound specifically to a subpopulation of photoreceptor cells and to discrete regions within the outer synaptic layer. The pattern of PNA binding suggests that this lectin binds preferentially to cone photoreceptor inner and outer segments and cone synaptic pedicles rather than to rod photoreceptor cells. This marked specificity of PNA binding suggests that it may provide a basis for the physical separation of cone and rod photoreceptor cells.
使用一组具有不同碳水化合物特异性的八种凝集素来研究小鼠视网膜细胞分化过程中糖缀合物表达的变化。所测试的凝集素包括伴刀豆球蛋白A(Con A)、麦胚凝集素(WGA)、大豆凝集素(SBA)、花生凝集素(PNA)、欧洲荆豆凝集素(UEA)、蓖麻凝集素I(RCA)、双花扁豆凝集素(DBA)和鲎试剂凝集素(LPA)。用异硫氰酸荧光素偶联的凝集素处理成年和出生后早期小鼠视网膜的未固定冰冻切片,并通过荧光显微镜检查。结果显示,在成年小鼠的细胞和突触视网膜层以及整个出生后发育过程中,凝集素存在选择性结合。一般来说,在视网膜发育过程中,观察到Con A、WGA、DBA、LPA、RCA和PNA的荧光凝集素染色强度增加。这表明在发育过程中碳水化合物部分的表达或可及性增加。SBA和UEA对成年或新生视网膜几乎没有或没有结合。在发育过程中和成年期,视网膜血管都被RCA强烈染色。所有与成年或新生视网膜层结合的凝集素都与光感受器细胞的内段区域有一定程度的反应性。然而,只有Con A、PNA和WGA与光感受器外段结合,这表明内段和外段膜的糖基化成分存在显著差异。PNA特异性结合光感受器细胞的一个亚群以及外突触层内的离散区域。PNA结合模式表明,这种凝集素优先结合视锥光感受器的内段和外段以及视锥突触小足,而不是视杆光感受器细胞。PNA结合的这种显著特异性表明,它可能为视锥和视杆光感受器细胞的物理分离提供基础。