Krivko I M, Rusakov D A, Savina S V, Skibo G G, Berezin V A
Dniepropetrovsk State University, Ukraine.
Neuroscience. 1993 Jul;55(2):491-8. doi: 10.1016/0306-4522(93)90518-k.
In monolayer cultures of hippocampal neurons from newborn rats, an immunocytochemical quantitative study was carried out to investigate age-dependent arrangement of the neural cell adhesion molecules in different parts of cell membranes. On the fifth and 12th day in vitro, neural cell adhesion molecules were labelled with specific antibodies and protein A conjugated to colloidal gold particles. Samples of randomly selected electron micrographs that displayed labelled membrane fragments of cell bodies, growth cones, and axons were numerically analysed for the five- and 12-day in vitro neurons. Neural cell adhesion molecules surface topography was quantitatively described and compared, using a statistical stereological approach. The mean surface density of labelled neural cell adhesion molecules was found to be approximately 2.5 times higher in growth cone membranes relative to somatic and axonal membranes in five-day in vitro neurons. By the 12th day in vitro, this density decreases in somatic membranes (approximately 18%) and increases in axonal membranes (approximately 60%). Representative spectra of lateral intervals between labels as well as images that show typical topography of label on membrane surfaces were simulated. The results revealed regular patterns of neural cell adhesion molecules on the somatic surface and allowed consideration of neural cell adhesion molecules arrangement in a view of membrane adhesion properties. Participation of cytoskeleton in neural cell adhesion molecules rearrangement is discussed.
在新生大鼠海马神经元的单层培养物中，进行了一项免疫细胞化学定量研究，以探讨神经细胞粘附分子在细胞膜不同部位的年龄依赖性排列。在体外培养的第5天和第12天，用特异性抗体和与胶体金颗粒偶联的蛋白A标记神经细胞粘附分子。对随机选择的显示细胞体、生长锥和轴突标记膜片段的电子显微镜图像样本，对体外培养5天和12天的神经元进行数值分析。使用统计体视学方法对神经细胞粘附分子的表面形貌进行定量描述和比较。发现在体外培养5天的神经元中，生长锥膜上标记的神经细胞粘附分子的平均表面密度相对于体细胞和轴突膜约高2.5倍。到体外培养第12天，这种密度在体细胞膜中降低（约18%），在轴突膜中增加（约60%）。模拟了标记之间横向间隔的代表性光谱以及显示膜表面标记典型形貌的图像。结果揭示了体细胞表面神经细胞粘附分子的规则模式，并从膜粘附特性的角度考虑了神经细胞粘附分子的排列。讨论了细胞骨架在神经细胞粘附分子重排中的作用。
