50B11 cells as a peripheral sensory nociceptive neuron model for neurotrophin signalling: Poster 62

Stimulation and sensitization of nociceptive neurons with cell bodies in dorsal root ganglia (DRG) play a critical role in the development of chronic and neuropathic pain.
Investigations of DRG neurons are limited by the absence of a suitable cellular high-throughput model capable of representing individual cell types found in DRG. The aim of this study was to establish whether an immortalized rat embryonic DRG cell (50B11), can be selectively differentiated into NGF-dependent peptidergic and GDNF-dependent non-peptidergic nociceptors. We used q-RT-PCR, Western blotting, In-Cell Western blots, immunohistochemistry and calcium efflux flow cytometry to determine the nociceptive characteristics of differentiated 50B11 cells via determination of markers at transcriptional (TRPV1, P2X3 and TrkA, n = 5) and translational levels (TRPV1,
CGRP, P2X3, TrkA, p75NTR, and GFRα, n = 5; immunohistochemistry, CGRP, SP, P2X3, and IB4, n = 5). Functional characteristics were investigated as response to ATP and capsaicin in 50B11 cells differentiated with forskolin in combination with NGF or GDNF. Our results show that NGF and GDNF treatment of 50B11 cells differentially regulate the cells to develop into a peptidergic (TRPV1 expressing, capsaicin-responsive) or non-peptidergic (P2X3 expressing, ATP-responsive) subtype with morphological features of peripheral sensory neurons. Our data indicate that the 50B11 cells – depending on culture conditions – have the capacity to differentiate into both peptidergic and non-peptidergic nociceptive neurons. This suggests that the cell line is a valuable tool for the study of nociceptive signaling events modeling peripheral nociceptors.