小脑颗粒神经元的培养。

CSH protocols Pub Date : 2008-12-01 DOI:10.1101/pdb.prot5107
Parizad M Bilimoria, Azad Bonni
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引用次数: 67

摘要

出生后大鼠小脑颗粒神经元的原代培养为神经元发育和功能的分子和细胞生物学研究提供了一个极好的模型系统。小脑皮层具有高度组织化的结构和很少的神经元亚型,提供了一个很好表征的神经回路。许多关于哺乳动物中枢神经系统中神经元凋亡、迁移和分化过程的基本见解都来自于对颗粒神经元的体外研究。颗粒神经元是大脑中最丰富的神经元类型。除了颗粒神经元的绝对体积之外,群体的同质性以及它们可以轻松转染的事实使它们成为阐明神经元发育的分子基础的理想选择。这种从产后大鼠中分离颗粒神经元的方法相对简单和快速,使用标准的酶解和机械解离方法。在含有有丝分裂抑制剂的血清培养基中,小脑颗粒神经元可以保持高纯度。轴突和树突可以根据形态和标记清楚地区分。为了更大的通用性,这种培养颗粒神经元的方案可以与基因敲除或转基因技术相结合,或用于小脑切片覆盖测定。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cultures of cerebellar granule neurons.

INTRODUCTIONPrimary cultures of granule neurons from the post-natal rat cerebellum provide an excellent model system for molecular and cell biological studies of neuronal development and function. The cerebellar cortex, with its highly organized structure and few neuronal subtypes, offers a well-characterized neural circuitry. Many fundamental insights into the processes of neuronal apoptosis, migration, and differentiation in the mammalian central nervous system have come from investigating granule neurons in vitro. Granule neurons are the most abundant type of neurons in the brain. In addition to the sheer volume of granule neurons, the homogeneity of the population and the fact that they can be transfected with ease render them ideal for elucidating the molecular basis of neuronal development. This protocol for isolating granule neurons from post-natal rats is relatively straightforward and quick, making use of standard enzymatic and mechanical dissociation methods. In a serum-based medium containing an inhibitor of mitosis, cerebellar granule neurons can be maintained with high purity. Axons and dendrites can be clearly distinguished on the basis of morphology and markers. For even greater versatility, this protocol for culturing granule neurons can be combined with knockout or transgenic technologies, or used in cerebellar slice overlay assays.

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