Santiago Marcelo F, Liour Sean S, Mendez-Otero Rosalia, Yu Robert K
Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
J Neurosci Res. 2005 Jul 1;81(1):9-20. doi: 10.1002/jnr.20532.
During development of the nervous system, neuronal precursors that originated in proliferative regions migrate along radial glial fibers to reach their final destination. P19 embryonal carcinoma (EC) stem cells exposed to retinoic acid (RA) differentiate into neurons, glia, and fibroblast-like cells. In this work, we induced P19 aggregates for 4 days with RA and plated them onto tissue culture dishes coated with poly-L-lysine. Several cells migrated out of and/or extended processes from the aggregates after 24 hr. Some cell processes were morphologically similar to radial glial fibers and stained for glial fibrillar acidic protein (GFAP) and nestin. Large numbers of migrating cells showed characteristics similar to those of bipolar migrating neurons and expressed the neuronal marker microtubule-associated protein 2. Furthermore, scanning electron microscopy analysis revealed an intimate association between the radial fibers and the migrating cells. Therefore, the migration of neuron-like cells on radial glia fibers in differentiated P19 aggregates resembled some of the migration models used thus far to study gliophilic neuronal migration. In addition, HPTLC analysis in this system showed the expression of 9-O-acetyl GD3, a ganglioside that has been associated with neuronal migration. Antibody perturbation assays showed that immunoblockage of 9-O-acetyl GD3 arrested neuronal migration in a reversible manner. In summary, we have characterized a new cell culture model for investigation of glial-guided neuronal migration and have shown that 9-O-acetyl GD3 ganglioside has an important role in this phenomenon.
在神经系统发育过程中，起源于增殖区域的神经元前体细胞沿着放射状胶质纤维迁移，以到达其最终目的地。暴露于视黄酸（RA）的P19胚胎癌细胞（EC）干细胞可分化为神经元、神经胶质细胞和成纤维细胞样细胞。在这项研究中，我们用RA诱导P19聚集体4天，然后将其接种到涂有聚-L-赖氨酸的组织培养皿上。24小时后，几个细胞从聚集体中迁移出来和/或伸出突起。一些细胞突起在形态上类似于放射状胶质纤维，并被胶质纤维酸性蛋白（GFAP）和巢蛋白染色。大量迁移细胞表现出与双极迁移神经元相似的特征，并表达神经元标志物微管相关蛋白2。此外，扫描电子显微镜分析显示放射状纤维与迁移细胞之间存在密切关联。因此，分化的P19聚集体中类神经元细胞在放射状胶质纤维上的迁移类似于迄今为止用于研究亲胶质神经元迁移的一些迁移模型。此外，该系统中的高效薄层层析分析显示了9-O-乙酰基GD3的表达，9-O-乙酰基GD3是一种与神经元迁移相关的神经节苷脂。抗体干扰试验表明，对9-O-乙酰基GD3的免疫阻断以可逆方式阻止了神经元迁移。总之，我们已经确定了一种用于研究胶质细胞引导的神经元迁移的新细胞培养模型，并表明9-O-乙酰基GD3神经节苷脂在这一现象中起重要作用。
