Isolation and culture of post-natal mouse cerebellar granule neuron progenitor cells and neurons.
Isolation and culture of post-natal mouse cerebellar granule neuron progenitor cells and neurons.
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DOI：
10.3791/990
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发表时间：
2009-01-16
期刊：
影响因子：
--
通讯作者：
Manzini, M Chiara
中科院分区：
文献类型：
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作者：
Lee, Hae Young;Greene, Lloyd A;Manzini, M Chiara
小脑皮质是一个很好的描述结构，为研究神经元的特性和发育提供了独特的机会。在小脑神经元类型（颗粒细胞、浦肯野细胞和抑制性中间神经元）中，颗粒神经元是迄今为止数量最多的，并且是哺乳动物脑中最丰富的神经元类型。在啮齿类动物中，小脑颗粒神经元在出生后的前两周内从小脑皮质最外层（外部颗粒层（EGL））的祖细胞产生。本文介绍了从出生后的小鼠小脑中富集和培养颗粒神经元及其祖细胞的技术。我们将描述程序，以获得培养物的纯度增加，可用于研究分化的祖细胞增殖成颗粒神经元。一旦祖细胞分化，培养物还提供了一个同质群体的颗粒神经元的实验操作和表征的现象，如突触，谷氨酸受体功能，与其他纯化的小脑细胞或细胞死亡的相互作用。
The cerebellar cortex is a well described structure that provides unique opportunities for studying neuronal properties and development. Of the cerebellar neuronal types (granule cells, Purkinje cells and inhibitory interneurons), granule neurons are by far the most numerous and are the most abundant type of neurons in the mammalian brain. In rodents, cerebellar granule neurons are generated during the first two post-natal weeks from progenitor cells in the outermost layer of the cerebellar cortex, the external granule layer (EGL). The protocol presented here describes techniques to enrich and culture granule neurons and their progenitor cells from post-natal mouse cerebellum. We will describe procedures to obtain cultures of increasing purity which can be used to study the differentiation of proliferating progenitor cells into granule neurons. Once the progenitor cells differentiate, the cultures also provide a homogenous population of granule neurons for experimental manipulation and characterization of phenomena such as synaptogenesis, glutamate receptor function, interaction with other purified cerebellar cells or cell death.