Abstract
The use of sensory neurons and assessment of neurite outgrowth in vitro is an important part of understanding neuronal development and plasticity. Cultures of rat dorsal root ganglion (DRG) neurons provide quantitative results very quickly and, when grown on growth promoting or inhibitory substrates, can be utilized to study axonal growth, neurotrophic dependence, and structure and function of growth cones. Since we are interested in axon regeneration and targeting, we have sought to promote neurite outgrowth by refining the techniques of growing DRG neurons in culture. This chapter describes detailed methods for the dissection and purification of DRG neurons and quantitative assessment of neurite on promoting or inhibitory substrates.
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References
Fitch MT, Silver J (2008) CNS injury, glial scars, and inflammation: inhibitory extracellular matrices and regeneration failure. Exp Neurol 209:294–301
Yiu G, He Z (2006) Glial inhibition of CNS axon regeneration. Nat Rev Neurosci 7:617–627
Sun F, He Z (2010) Neuronal intrinsic barriers for axon regeneration in the adult CNS. Curr Opin Neurobiol 20:510–518
Filbin MT (2003) Myelin-associated inhibitors of axonal regeneration in the adult mammalian CNS. Nat Rev Neurosci 4:703–713
Liu K, Lu Y, Lee JK, Samara R, Willenberg R, Sears-Kraxberger I et al (2010) PTEN deletion enhances the regenerative ability of adult corticospinal neurons. Nat Neurosci 13:1075–1081
Qiu J, Cai D, Dai H, McAtee M, Hoffman PN, Bregman BS et al (2002) Spinal axon regeneration induced by elevation of cyclic AMP. Neuron 34:895–903
Neumann S, Bradke F, Tessier-Lavigne M, Basbaum AI (2002) Regeneration of sensory axons within the injured spinal cord induced by intraganglionic cAMP elevation. Neuron 34:885–893
Park KK, Liu K, Hu Y, Smith PD, Wang C, Cai B et al (2008) Promoting axon regeneration in the adult CNS by modulation of the PTEN/mTOR pathway. Science 322:963–966
Moore DL, Blackmore MG, Hu Y, Kaestner KH, Bixby JL, Lemmon VP et al (2009) KLF family members regulate intrinsic axon regeneration ability. Science 326:298–301
Qiu J, Cafferty WB, McMahon SB, Thompson SW (2005) Conditioning injury-induced spinal axon regeneration requires signal transducer and activator of transcription 3 activation. J Neurosci 25:1645–1653
Snow DM, Smith JD, Cunningham AT, McFarlin J, Goshorn EC (2003) Neurite elongation on chondroitin sulfate proteoglycans is characterized by axonal fasciculation. / Exper. Neurology 182(2003):310–321
Acknowledgments
This work was supported by NIH grant RO1 NS060784, Shriners Hospitals for Children (GMS) and Shriners Postdoctoral Fellowship (GMS and YL).
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Smith, G.M., Liu, Y. (2021). Quantitative Assessment of Neurite Outgrowth Over Growth Promoting or Inhibitory Substrates. 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_13
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DOI: https://doi.org/10.1007/978-1-0716-1437-2_13
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Publisher Name: Humana, New York, NY
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