Myelination of rodent hippocampal neurons in culture.

Axons of various hippocampal neurons are myelinated mainly postnatally, which is important for the proper function of neural circuits. Demyelination in the hippocampus has been observed in patients with multiple sclerosis, Alzheimer’s disease or temporal lobe epilepsy. However, very little is known about the mechanisms and exact functions of the interaction between the myelin-making oligodendrocytes and the axons within the hippocampus. This is mainly attributable to the lack of a system suitable for molecular studies. We recently established a new myelin coculture from embryonic day (E) 18 rat embryos consisting of hippocampal neurons and oligodendrocytes, with which we identified a novel intra-axonal signaling pathway regulating the juxtaparanodal clustering of Kv1.2 channels. Here we describe the detailed protocol for this new coculture. It takes about 5 weeks to set up and use the system. This coculture is particularly useful for studying myelin-mediated regulation of ion channel trafficking and for understanding how neuronal excitability and synaptic transmission are regulated by myelination.

各种海马神经元的轴突主要在出生后形成髓鞘，这对神经回路的正常功能至关重要。在多发性硬化症、阿尔茨海默病或颞叶癫痫患者中已观察到海马体脱髓鞘现象。然而，对于制造髓鞘的少突胶质细胞与海马体内轴突之间相互作用的机制和确切功能知之甚少。这主要是由于缺乏适合分子研究的系统。我们最近从胚胎第18天（E18）的大鼠胚胎建立了一种新的髓鞘共培养体系，由海马神经元和少突胶质细胞组成，利用该体系我们确定了一种调节Kv1.2通道近节旁聚集的新型轴突内信号通路。在此我们描述这种新的共培养体系的详细实验方案。建立和使用该系统大约需要5周时间。这种共培养体系对于研究髓鞘介导的离子通道运输调节以及理解神经元兴奋性和突触传递如何受髓鞘形成调节特别有用。