Abstract
Preparations of peripheral sensory neurons from rodents are essential for studying the molecular mechanism of neuronal survival and physiology. Although, isolating and culturing these neurons proves difficult, often these preparations are contaminated with nonneuronal proliferating cells. Here, we describe an isolation method using a Percoll gradient and an antimitotic reagent to significantly reduce the nonneuronal cell contamination while maintaining the integrity of the rodent sensory dorsal root ganglia (DRG) neurons.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Woolf CJ, Ma Q (2007) Nociceptors—noxious stimulus detectors. Neuron 55(3):353–364
Caspary T, Anderson KV (2003) Patterning cell types in the dorsal spinal cord: what the mouse mutants say. Nat Rev Neurosci 4(4):289–297
Vikman KS, Kristensson K, Hill RH (2001) Sensitization of dorsal horn neurons in a two-compartment cell culture model: wind-up and long-term potentiation-like responses. J Neurosci 21(19):RC169
Bolyard LA, Van Looy JW, Vasko MR (2000) Sensitization of rat sensory neurons by chronic exposure to forskolin or ‘inflammatory cocktail’ does not downregulate and requires continuous exposure. Pain 88(3):277–285
Rich KM, Disch SP, Eichler ME (1989) The influence of regeneration and nerve growth factor on the neuronal cell body reaction to injury. J Neurocytol 18(5):569–576
Keynes R et al (1997) Surround repulsion of spinal sensory axons in higher vertebrate embryos. Neuron 18(6):889–897
Hari A et al (2004) Neurotrophins and extracellular matrix molecules modulate sensory axon outgrowth. Int J Dev Neurosci 22(2):113–117
Watanabe K et al (2006) Dorsally derived netrin 1 provides an inhibitory cue and elaborates the ‘waiting period’ for primary sensory axons in the developing spinal cord. Development 133(7):1379–1387
Sainath R, Gallo G (2015) The dynein inhibitor Ciliobrevin D inhibits the bidirectional transport of organelles along sensory axons and impairs NGF-mediated regulation of growth cones and axon branches. Dev Neurobiol 75(7):757–777
Bray D, Thomas C, Shaw G (1978) Growth cone formation in cultures of sensory neurons. Proc Natl Acad Sci U S A 75(10):5226–5229
Lamoureux P, Buxbaum RE, Heidemann SR (1998) Axonal outgrowth of cultured neurons is not limited by growth cone competition. J Cell Sci 111(Pt 21):3245–3252
Burgess GM et al (1989) Second messengers involved in the mechanism of action of bradykinin in sensory neurons in culture. J Neurosci 9(9):3314–3325
Adler JE, Walker PD (2000) Cyclic AMP regulates substance P expression in developing and mature spinal sensory neurons. J Neurosci Res 59(5):624–631
Crawford AT et al (2008) E-cadherin expression in postnatal Schwann cells is regulated by the cAMP-dependent protein kinase a pathway. Glia 56(15):1637–1647
Camarena V et al (2010) Nature and duration of growth factor signaling through receptor tyrosine kinases regulates HSV-1 latency in neurons. Cell Host Microbe 8(4):320–330
Eichler ME, Rich KM (1989) Death of sensory ganglion neurons after acute withdrawal of nerve growth factor in dissociated cell cultures. Brain Res 482(2):340–346
Acknowledgments
This work is supported by an NIH grant R01NS082116 (THB) and the Comprehensive NeuroAIDS Center grant P30MH092 I 777 (MS).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Shekarabi, M., Robinson, J.A., Burdo, T.H. (2021). Isolation and Culture of Dorsal Root Ganglia (DRG) from Rodents. In: Amini, S., White, M.K. (eds) Neuronal Cell Culture. Methods in Molecular Biology, vol 2311. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1437-2_14
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1437-2_14
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1436-5
Online ISBN: 978-1-0716-1437-2
eBook Packages: Springer Protocols