Labeling of Precursor Granule Cells in the Cerebellum by ex vivo Electroporation
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Author:
Date:
2013-06-20
[Abstract] This protocol will be useful to introduce the genes of interest into the cerebellar granule cells at early stages of development. Since the granule cell precursors are localized in the external granule layer before migration, DNA plasmids can be specifically incorporated into the granule cells by injecting DNA solution into the cerebellar fissures followed by application of electric pulses. This technique can be performed prior to the preparation of either dissociated or organotypic culture, which can be used to study the molecular mechanisms of cell migration, axon elongation and synapstogenesis during development.
[摘要] 该方案可用于在发育的早期阶段将感兴趣的基因引入小脑颗粒细胞。 由于颗粒细胞前体在迁移前位于外部颗粒层中,因此通过将DNA溶液注射到小脑裂隙中,然后施加电脉冲,可将DNA质粒特异性地掺入颗粒细胞中。 该技术可以在制备解离或器官型培养物之前进行,其可以用于研究在发育期间细胞迁移,轴突伸长和突触发生的分子机制。
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In utero Electroporation of Mouse Embryo Brains
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Author:
Date:
2012-07-20
[Abstract] This is a non-invasive technique to introduce transgenes into developing brains. In this technique, DNA is injected into the lateral ventricle of the embryonic brains, and is incorporated into the cells through electroporation. Embryos then continue their development in normal conditions in vivo. The effects of genes of interest can be evaluated at certain time points after in utero electroporation. This technique allows acute knockdown or over expression of genes of interest. Compensatory effects from other genes are less likely to happen; it also circumvents possible chronic detrimental effects.
[摘要] 这是将转基因引入发育中的大脑的非侵入性技术。 在该技术中,将DNA注入胚胎脑的侧脑室,并通过电穿孔掺入细胞中。 胚胎然后在正常条件下在体内继续其发育。 可以在子宫内电穿孔后的某些时间点评估感兴趣的基因的效应。 这种技术允许急性敲减或过表达感兴趣的基因。 其他基因的补偿效应不太可能发生; 它也绕过可能的慢性有害影响。
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