Immortalization and characterization of a nociceptive dorsal root ganglion sensory neuronal line.
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.
Chen
Weiran
W
Department of Neurology, School of Medicine, Johns Hopkins University, Baltimore, MD, USA.
Mi
Ruifa
R
Haughey
Norman
N
Oz
Murat
M
Höke
Ahmet
A
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002454:Cell Differentiation; D002460:Cell Line; D004353:Drug Evaluation, Preclinical; D018274:Electroporation; D005727:Ganglia, Spinal / Q000166:cytology*; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D009474:Neurons / Q000166:cytology* / Q000502:physiology*; D018696:Neuroprotective Agents / Q000494:pharmacology; D009619:Nociceptors / Q000166:cytology* / Q000502:physiology*; D016133:Polymerase Chain Reaction; D051381:Rats; D050916:TRPV Cation Channels / Q000096:biosynthesis; D019098:Telomerase / Q000235:genetics
D000952:Antigens, Polyomavirus Transforming; D018696:Neuroprotective Agents; D050916:TRPV Cation Channels; C493922:Trpv1 protein, rat; D019098:Telomerase