Department of Cancer Biology and Wake Forest Baptist Comprehensive Cancer Center, Wake Forest University Health Sciences, Winston-Salem, NC 27157, USA.
Department of Anesthesiology, Wake Forest University Health Sciences, Winston-Salem, NC 27157, USA.
Biochem Pharmacol. 2021 Jun;188:114520. doi: 10.1016/j.bcp.2021.114520. Epub 2021 Mar 17.
Abnormal outgrowth of sensory nerves is one of the important contributors to pain associated with cancer and its treatments. Primary neuronal cultures derived from dorsal root ganglia (DRG) have been widely used to study pain-associated signal transduction and electrical activity of sensory nerves. However, there are only a few studies using primary DRG neuronal culture to investigate neurite outgrowth alterations due to underlying cancer-related factors and chemotherapeutic agents. In this study, primary DRG sensory neurons derived from mouse, non-human primate, and human were established in serum and growth factor-free conditions. A bovine serum albumin gradient centrifugation method improved the separation of sensory neurons from satellite cells. The purified DRG neurons were able to maintain their heterogeneous subpopulations, and displayed an increase in neurite growth when exposed to cancer-derived conditioned medium, while they showed a reduction in neurite length when treated with a neurotoxic chemotherapeutic agent. Additionally, a semi-automated quantification method was developed to measure neurite length in an accurate and time-efficient manner. Finally, these exogenous factors altered the gene expression patterns of murine primary sensory neurons, which are related to nerve growth, and neuro-inflammatory pain and nociceptor development. Together, the primary DRG neuronal culture in combination with a semi-automated quantification method can be a useful tool for further understanding the impact of exogenous factors on the growth of sensory nerve fibers and gene expression changes in sensory neurons.
感觉神经的异常生长是与癌症及其治疗相关的疼痛的重要原因之一。源自背根神经节（DRG）的原代神经元培养物已广泛用于研究与疼痛相关的信号转导和感觉神经的电活动。然而，仅有少数研究使用原代 DRG 神经元培养物来研究由于潜在的癌症相关因素和化疗药物引起的轴突生长改变。在这项研究中，从小鼠、非人类灵长类动物和人类中建立了在无血清和生长因子条件下的原代 DRG 感觉神经元。牛血清白蛋白梯度离心法改善了感觉神经元与卫星细胞的分离。纯化的 DRG 神经元能够维持其异质亚群，并且在暴露于癌症衍生的条件培养基时表现出轴突生长增加，而在用神经毒性化疗药物处理时则表现出轴突长度减少。此外，还开发了一种半自动定量方法，以准确且高效的方式测量轴突长度。最后，这些外源性因素改变了与神经生长、神经炎性疼痛和伤害感受器发育相关的小鼠原代感觉神经元的基因表达模式。总之，结合半自动定量方法的原代 DRG 神经元培养物可以成为进一步了解外源性因素对感觉神经纤维生长和感觉神经元基因表达变化影响的有用工具。
