Differentiation of the 50B11 dorsal root ganglion cells into NGF and GDNF responsive nociceptor subtypes
Authors: Marri Shashikanth
Best G Oliver
Wei Yingkai
Matusica Dusan
Voelcker H Nicolas
Martin M Alyce
Erickson Andelain
Canlas Jastrow
Haberberger V Rainer
Brierley M Stuart
Barry M Christine
Michael Z Michael
Keating J Damien
Journal: Molecular Pain (2020)
Discipline: NEUROSCIENCE & BEHAVIOR
Keywords: 50B11; ATP; RNA-seq; calcium activity; capsaicin
Affiliations: Anatomy and Histology, Flinders Health & Medical Research Institute, College of Medicine and Public Health, Flinders University, Adelaide, South Australia, Australia., Human Physiology, Flinders Health & Medical Research Institute, College of Medicine and Public Health, Flinders University, Adelaide, South Australia, Australia., Flinders Centre for Innovation in Cancer, College of Medicine and Public Health, Flinders University and Medical Centre, Bedford Park, Adelaide, South Australia, Australia., Visceral Pain Research Group, College of Medicine and Public Health, South Australian Health and Medical Research Institute, Flinders University, Adelaide, South Australia, Australia., Flow Cytometry Facility, Department of Molecular Medicine and Genetics, College of Medicine and Public Health, Flinders University, Adelaide, South Australia, Australia.

Abstract:  The embryonic rat dorsal root ganglion (DRG) neuron-derived 50B11 cell line is a promising sensory neuron model expressing markers characteristic of NGF and GDNF-dependent C-fibre nociceptors. Whether these cells have the capacity to develop into distinct nociceptive subtypes based on NGF- or GDNF-dependence has not been investigated. Here we show that by augmenting forskolin (FSK) and growth factor supplementation with NGF or GDNF, 50B11 cultures can be driven to acquire differential functional responses to common nociceptive agonists capsaicin and ATP respectively. In addition, to previous studies, we also demonstrate that a differentiated neuronal phenotype can be maintained for up to 7 days. Western blot analysis of nociceptive marker proteins further demonstrates that the 50B11 cells partially recapitulate the functional phenotypes of classical NGF-dependent (peptidergic) and GDNF-dependent (non-peptidergic) neuronal subtypes described in DRGs. Further, 50B11 cells differentiated with NGF/FSK, but not GDNF/FSK, show sensitization to acute prostaglandin E2 treatment. Finally, RNA-Seq analysis confirms that differentiation with NGF/FSK or GDNF/FSK produces two 50B11 cell subtypes with distinct transcriptome expression profiles. Gene ontology comparison of the two subtypes of differentiated 50B11 cells to rodent DRG neurons studies shows significant overlap in matching or partially matching categories. This transcriptomic analysis will aid future suitability assessment of the 50B11 cells as a high-throughput nociceptor model for a broad range of experimental applications. In conclusion, this study shows that the 50B11 cell line is capable of partially recapitulating features of two distinct types of embryonic NGF and GDNF-dependent nociceptor-like cells. 

Retracted: No

PubPeer comments:
 Figure 4 western blot - manipulated beta 3 tubulin bands throughout. Very obvious , 
 This paper has been incorrectly abstracted and indexed by PubMed because its author list inverts the first and last name of the authors. So for example it is listing  Dusan M. as the first author when it should be  Matusica D. . ,