College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
Anal Chem. 2012 Aug 7;84(15):6444-53. doi: 10.1021/ac3013708. Epub 2012 Jul 26.
Studies on the degeneration and regeneration of neurons as individual compartments of axons or somata can provide critical information for the clinical therapy of nervous system diseases. A controllable in vitro platform for multiple purposes is key to such studies. In the present study, we describe an integrated microfluidic device designed for achieving localized stimulation to neuronal axons or somata. We observed neuronal compartment degeneration after localized chemical stimulation and regeneration under the accessorial function of an interesting compound treatment or coculture with desired cells in controllable chambers. In a spatiotemporally controlled manner, this device was used to investigate hippocampal neuronal soma and axon degeneration after acrylamide stimulation, as well as subsequent regeneration after treatment with the monosialoganglioside GM1 or with cocultured glial cells (astrocytes or Schwann cells). To gain insight into the molecular mechanisms that mediate neuronal injury and regeneration, as well as to investigate whether acrylamide stimulation to neurons induces changes in Ca(2+) concentrations, the related neuronal genes and real-time Ca(2+) signal in neurons were also analyzed. The results showed that neuronal axons were more resistant to acrylamide injury than neuronal somata. Under localized stimulation, axons had self-destruct programs different from somata, and somatic injury caused the secondary response of axon collapse. This study provides a foundation for future in-depth analyses of spatiotemporally controlled and multifactor neuronal compartment regeneration after various injuries. The microfluidic device is also useful in evaluating potential therapeutic strategies to treat chemical injuries involving the central nervous system.
作为轴突或胞体的神经元的变性和再生的研究可以为神经系统疾病的临床治疗提供关键信息。一个用于多种目的的可控体外平台是此类研究的关键。在本研究中，我们描述了一种集成的微流控设备，用于实现对神经元轴突或胞体的局部刺激。我们观察到局部化学刺激后神经元隔室的变性，以及在有趣的化合物处理或与所需细胞共培养的辅助功能下在可控腔室内的再生。以时空可控的方式，该设备用于研究丙烯酰胺刺激后海马神经元胞体和轴突的变性，以及随后用单唾液酸神经节苷脂 GM1 或与共培养的神经胶质细胞（星形胶质细胞或施万细胞）处理后的再生。为了深入了解介导神经元损伤和再生的分子机制，并研究丙烯酰胺刺激神经元是否会引起细胞内钙离子浓度的变化，还分析了相关的神经元基因和神经元内的实时钙离子信号。结果表明，神经元轴突对丙烯酰胺损伤的抵抗力强于神经元胞体。在局部刺激下，轴突具有不同于胞体的自我毁灭程序，而胞体损伤引起轴突崩溃的继发性反应。这项研究为未来深入分析各种损伤后时空可控和多因素神经元隔室再生奠定了基础。该微流控设备在评估潜在的治疗策略以治疗涉及中枢神经系统的化学损伤方面也很有用。
