Falk Torsten, Zhang Shiling, Erbe Emilie L, Sherman Scott J
Department of Neurology, The University of Arizona, Tucson, 85724-5023, USA.
J Comp Neurol. 2006 Jan 10;494(2):275-89. doi: 10.1002/cne.20819.
Neurons maintained in dispersed primary culture offer a number of advantages as a model system and are particularly well-suited for studies of the intrinsic electrical properties of neurons by patch clamp. We have characterized the immunocytochemical and electrophysiological properties of cultured rat striatal neurons as they develop in vitro in order to compare this model system with the known properties found in vivo. We found a high abundance of cells in vitro corresponding to the principal striatal output neuron, the medium spiny neuron. Immunocytochemical studies indicate that these cells have both dopamine-1 and dopamine-2 receptors and that there is overlap in their expression within the population of neurons. Semiquantitative analysis revealed bimodal distributions of dopamine receptor expression among the population of neurons. The principal peptide neurotransmitters substance P and enkephalin were present but at reduced levels compared with adult preparations. Other striatal markers such as calbindin, calretinin, and the cannabinoid-1 receptor were abundant. An immunocytochemical survey of voltage-gated K(+) channel subunits characteristic of adult tissue demonstrated the presence in vitro of Kv1.1, Kv1.4, Kv4.2, Kv4.3, and Kvbeta1.1, which have been associated with the rapidly inactivating currents. Electrophysiological studies employing voltage clamp revealed that outward currents had a large inactivating (A-type) component characteristic of mature basal ganglia. Current clamp studies reveal complex spontaneous firing patterns in a subset of neurons, including bursting behaviors superimposed on a slow depolarization. The inward rectifying channels Kir2.1 and Kir2.3, which are specific to particular compartments in adult striatum, were present in culture.
原代分散培养的神经元作为一种模型系统具有诸多优势,尤其适合通过膜片钳研究神经元的内在电特性。我们已经对培养的大鼠纹状体神经元在体外发育过程中的免疫细胞化学和电生理特性进行了表征,以便将这个模型系统与体内已知特性进行比较。我们发现体外有大量细胞对应于纹状体主要输出神经元,即中等棘状神经元。免疫细胞化学研究表明,这些细胞同时具有多巴胺1型和多巴胺2型受体,并且它们在神经元群体中的表达存在重叠。半定量分析揭示了神经元群体中多巴胺受体表达的双峰分布。主要肽类神经递质P物质和脑啡肽存在,但与成年组织相比水平降低。其他纹状体标志物如钙结合蛋白、钙视网膜蛋白和大麻素1型受体含量丰富。对成年组织特有的电压门控钾离子通道亚基进行的免疫细胞化学调查表明,体外存在Kv1.1、Kv1.4、Kv4.2、Kv4.3和Kvbeta1.1,这些与快速失活电流有关。采用电压钳的电生理研究表明,外向电流具有成熟基底神经节特有的大失活(A型)成分。电流钳研究揭示了一部分神经元中复杂的自发放电模式,包括叠加在缓慢去极化上的爆发行为。成年纹状体特定区域特有的内向整流通道Kir2.1和Kir2.3在培养物中也存在。