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使用病毒载体将转基因体内导入小鼠坐骨神经细胞

In Vivo Introduction of Transgenes into Mouse Sciatic Nerve Cells Using Viral Vectors.

作者信息

Van Hameren Gerben, Gonzalez Sergio, Fernando Ruani N, Perrin-Tricaud Claire, Tricaud Nicolas

机构信息

Institut des Neurosciences de Montpellier, INSERM U1051, Université de Montpellier, Montpellier, France.

出版信息

Methods Mol Biol. 2018;1791:263-276. doi: 10.1007/978-1-4939-7862-5_21.

Abstract

Myelinated fibers are essential for the rapid and efficient propagation of nerve information throughout the body. These fibers result from an intimate crosstalk between myelinating glia and the myelinated axons and, because it is difficult to fully reproduce these interactions in vitro, the basic molecular mechanisms that regulate myelination, demyelination, and remyelination remain unclear. Schwann cells produce myelin in the peripheral nervous system (PNS) and remain associated with the axons of peripheral neurons throughout axonal migration to the target. In order to investigate more closely the biology of myelinated fibers, we developed a local transgenesis approach based on the injection of engineered viral vectors in the sciatic nerve of mice to locally transduce peripheral nerve cells. This approach represents an alternative to germline modifications as it facilitates and speed up the investigation of peripheral nerve biology in vivo. Indeed the protocol we describe here requires just 3 weeks to complete. The injection of engineered viral vectors in the sciatic nerve of mice is a reproducible and straightforward method for introducing exogenous factors into myelinating Schwann cells and myelinated axons in vivo in order to investigate specific molecular mechanisms.

摘要

有髓纤维对于神经信息在全身的快速高效传播至关重要。这些纤维源于髓鞘形成神经胶质细胞与有髓轴突之间密切的相互作用,并且由于难以在体外完全重现这些相互作用,调节髓鞘形成、脱髓鞘和髓鞘再生的基本分子机制仍不清楚。施万细胞在周围神经系统(PNS)中产生髓鞘,并在轴突向靶标迁移的整个过程中与周围神经元的轴突保持关联。为了更深入地研究有髓纤维的生物学特性,我们开发了一种局部转基因方法,该方法基于向小鼠坐骨神经注射工程化病毒载体以局部转导周围神经细胞。这种方法是种系修饰的一种替代方法,因为它有助于并加速体内周围神经生物学的研究。实际上,我们在此描述的方案仅需3周即可完成。向小鼠坐骨神经注射工程化病毒载体是一种可重复且直接的方法,用于在体内将外源性因子引入形成髓鞘的施万细胞和有髓轴突中,以研究特定的分子机制。

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