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利用实时聚合酶链反应研究中枢神经系统中的髓磷脂基因表达

Study of Myelin Gene Expression in the Central Nervous System Using Real-Time PCR.

作者信息

El Khoury Diala

机构信息

Department of Biology Louaize Lebanon, NDU Natural and Applied Sciences, Notre Dame University, Zouk Mosbeh, Lebanon.

出版信息

Methods Mol Biol. 2019;2011:659-670. doi: 10.1007/978-1-4939-9554-7_38.

Abstract

Myelin sheaths are crucial for the survival and maintenance of the axons and the rapid propagation of the action potential. The glial cells involved are Schwann cells in the peripheral nervous system (PNS) and oligodendrocytes in the central nervous system (CNS). One oligodendrocyte may myelinate over 40 axons. In the CNS, myelin is composed of several layers of cytoplasmic membrane from oligodendrocytes stabilized by structural myelin-specific proteins such as proteolipid protein (PLP) and myelin basic protein (MBP). Those genes are expressed during myelination and then silenced. They can be re-expressed after demyelinating episodes, where they contribute to remyelination. Demyelination occurs after injuries of the CNS such as traumatic brain injury or during acute episodes of neurodegeneration observed in demyelinating and neurodegenerative diseases. Remyelination process is achieved by oligodendrocytes newly generated following the recruitment and differentiation of oligodendrocyte precursor cells (OPCs). Failure of remyelination process leads to irreversible axonal loss, functional impairment, and finally decreased cognitive performances. Several techniques have been described to study myelination and remyelination in culture systems. In this chapter, we explain how we can study myelin genes' expression in oligodendrocytes by real-time polymerase chain reaction (RT-PCR) using specific primers for plp and mbp. This technique can be crucial and prompt to determine the effect of specific chemicals (such as pesticides) on the myelination process in oligodendrocytes.

摘要

髓鞘对于轴突的存活和维持以及动作电位的快速传播至关重要。所涉及的神经胶质细胞在外周神经系统(PNS)中是施万细胞,在中枢神经系统(CNS)中是少突胶质细胞。一个少突胶质细胞可能会使40多条轴突形成髓鞘。在中枢神经系统中,髓鞘由少突胶质细胞的几层细胞质膜组成,这些膜由结构髓鞘特异性蛋白如蛋白脂蛋白(PLP)和髓鞘碱性蛋白(MBP)稳定。这些基因在髓鞘形成过程中表达,然后沉默。它们可以在脱髓鞘发作后重新表达,在脱髓鞘发作中它们有助于再髓鞘化。脱髓鞘发生在中枢神经系统损伤后,如创伤性脑损伤,或在脱髓鞘和神经退行性疾病中观察到的神经退行性急性发作期间。再髓鞘化过程是由少突胶质细胞前体细胞(OPC)募集和分化后新产生的少突胶质细胞实现的。再髓鞘化过程失败会导致不可逆的轴突损失、功能障碍,最终导致认知能力下降。已经描述了几种技术来研究培养系统中的髓鞘形成和再髓鞘化。在本章中,我们解释了如何使用针对plp和mbp的特异性引物,通过实时聚合酶链反应(RT-PCR)来研究少突胶质细胞中髓鞘基因的表达。这项技术对于确定特定化学物质(如农药)对少突胶质细胞髓鞘形成过程的影响可能至关重要且迅速。

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