Department of Neurology, Medical Faculty, Research Group for Clinical and Experimental Neuroimmunology, Heinrich-Heine-University, Düsseldorf, Germany.
J Neurosci Methods. 2013 Mar 30;214(1):69-79. doi: 10.1016/j.jneumeth.2013.01.009. Epub 2013 Jan 21.
The rat dorsal root ganglia (DRG) model is a long-standing in vitro model for analysis of myelination in the peripheral nervous system. For performing systematic, high throughput analysis with transgenic animals, a simplified BL6 mouse protocol is indispensable. Here we present a stable and reliable protocol for myelinating co-cultures producing a high myelin ratio using cells from C57BL/6 mice. As an easy accessible and operable method, Sudan staining proved to be efficient in myelin detection for fixed cultures. Green fatty acid stain turned out to be highly reliable for analysis of the dynamic biological processes of myelination in vital cultures. Once myelinated we were able to induce demyelination by the addition of forskolin into the model system. In addition, we provide an optimised rat DRG protocol with significantly improved myelin ratio and a comparison of the protocols presented. Our results strengthen the value of ex vivo myelination models in neurobiology.
大鼠背根神经节（DRG）模型是一种长期存在的体外模型，可用于分析周围神经系统中的髓鞘形成。对于使用转基因动物进行系统的高通量分析，简化的 BL6 小鼠方案是必不可少的。在这里，我们提出了一种稳定可靠的髓鞘形成共培养方案，使用 C57BL/6 小鼠的细胞产生高髓鞘比。作为一种易于操作的方法，苏丹染色在固定培养物中的髓鞘检测中非常有效。绿色脂肪酸染色在活细胞中分析髓鞘形成的动态生物学过程中非常可靠。一旦髓鞘化，我们就可以通过向模型系统中添加 forskolin 来诱导脱髓鞘。此外，我们还提供了一种优化的大鼠 DRG 方案，该方案具有显著提高的髓鞘比，并对所提出的方案进行了比较。我们的结果增强了离体髓鞘形成模型在神经生物学中的价值。
