Crawford G D, Le W D, Smith R G, Xie W J, Stefani E, Appel S H
Department of Neurology, Baylor College of Medicine, Houston, Texas 77030.
J Neurosci. 1992 Sep;12(9):3392-8. doi: 10.1523/JNEUROSCI.12-09-03392.1992.
A dopaminergic neuroblastoma was derived using somatic cell fusion of rat embryonic mesencephalon cells and the murine neuroblastoma-glioma cell line N18TG2. The resulting interspecies hybrid, named MES23.5, has retained a stable phenotype and karyotype for a continuous culture period of 1 year. The hybrid exhibits several properties that suggest that the parent primary neurons originated in the substantia nigra. The cell line contains tyrosine hydroxylase, which is identifiable both by biochemical and immunological methods and synthesizes dopamine, but no other catecholamine. Additionally, the cell line expresses apparent voltage-gated CA2+ channels as measured by high-affinity omega-conotoxin binding. The MES23.5 omega-conotoxin receptors are of similar affinity class to those found in adult rat mesencephalon. No dihydropyridine receptors, as measured by PN200-100 ligand binding, are present. None of these properties are found in the N18TG2 parent. At least three neuronal features, namely, tyrosine hydroxylase, dopamine synthesis, and omega-conotoxin receptor expression, are quantitatively elevated after sustained treatment with cAMP analogs. The cell line expresses a complex range of neural properties found in the dopaminergic neurons of the substantia nigra, and may therefore be useful elucidating further details of their cell biology.
通过大鼠胚胎中脑细胞与小鼠神经母细胞瘤 - 胶质瘤细胞系N18TG2的体细胞融合获得了一种多巴胺能神经母细胞瘤。产生的种间杂种命名为MES23.5,在连续培养1年的时间里保持了稳定的表型和核型。该杂种表现出的一些特性表明其亲本原代神经元起源于黑质。该细胞系含有酪氨酸羟化酶,可通过生化和免疫方法鉴定,并且能合成多巴胺,但不合成其他儿茶酚胺。此外,通过高亲和力ω - 芋螺毒素结合测量发现该细胞系表达明显的电压门控Ca2 +通道。MES23.5的ω - 芋螺毒素受体与成年大鼠中脑中发现的受体具有相似的亲和力类别。通过PN200 - 100配体结合测量,不存在二氢吡啶受体。这些特性在N18TG2亲本中均未发现。在用cAMP类似物持续处理后,至少三种神经元特征,即酪氨酸羟化酶、多巴胺合成和ω - 芋螺毒素受体表达在数量上有所增加。该细胞系表达了黑质多巴胺能神经元中发现的一系列复杂的神经特性,因此可能有助于阐明其细胞生物学的更多细节。