Immortalization and characterization of a nociceptive dorsal root ganglion sensory neuronal line

被引:86
作者
Chen, Weiran
Mi, Ruifa
Haughey, Norman
Oz, Murat
Hoeke, Ahmet
机构
[1] Johns Hopkins Univ, Sch Med, Dept Neurol, Baltimore, MD 21205 USA
[2] Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21205 USA
[3] NIDA, Integrat Neurosci Sect, Intramural Res Program, NIH, Baltimore, MD USA
关键词
DRG; high-throughput screen; nociceptive; sensory neuronal line; TRPV-1; 50B11;
D O I
10.1111/j.1529-8027.2007.00131.x
中图分类号
R74 [神经病学与精神病学];
学科分类号
摘要
Development of neuroprotective strategies for peripheral neuropathies requires high-throughput drug screening assays with appropriate cell types. Currently, immortalized dorsal root ganglion (DRG) sensory neuronal cell lines that maintain nociceptive sensory neuronal properties are not available. We generated immortalized DRG neuronal lines from embryonic day 14.5 rats. Here, we show that one of the immortalized DRG neuronal lines, 50B11, has the properties of a nociceptive neuron. When differentiated in the presence of forskolin, these cells extend long neurites, express neuronal markers, and generate action potentials. They express receptors and markers of small-diameter sensory neurons and upregulate appropriate receptor populations when grown in the presence of glial cell line-derived neurotrophic factor or nerve growth factor. Furthermore, they express capsaicin receptor transient receptor potential vanilloid family-1 (TRPV-1) and respond to capsaicin with increases in intracellular calcium. In a 96-well plate format, these neurons show a decline in ATP levels when exposed to dideoxycytosine (ddC) in a proper time- and dose-dependent manner. This ddC-induced reduction in ATP levels correlates with axonal degeneration. The immortalized DRG neuronal cell line 50B11 can be used for high-throughput drug screening for neuroprotective agents for axonal degeneration and antinociceptive drugs that block TRPV-1.
引用
收藏
页码:121 / 130
页数:10
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