Immunoablation of cells expressing the NG2 chondroitin sulphate proteoglycan.

Immunoablation of cells expressing the NG2 chondroitin sulphate proteoglycan.
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DOI:
10.1111/joa.12141
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发表时间:
2014-02
期刊:
影响因子:
2.4
通讯作者:
Butt AM
Butt AM
中科院分区:
医学3区
文献类型:
--
作者:
Leoni G;Rattray M;Fulton D;Rivera A;Butt AM

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跨膜的NG2硫酸软骨素蛋白多糖(CSPG)的表达定义了一个独特的NG2胶质细胞群体。NG2胶质细胞是成人中枢神经系统(CNS)中少突胶质前体细胞的再生池,对多发性硬化症等脱髓鞘疾病具有重要作用,也是CNS损伤后抑制轴突再生的胶质瘢痕的主要组成部分。此外,NG2-神经胶质细胞形成独特的神经元-神经胶质突触,功能未解。然而,到目前为止,使用传统的转基因NG2‘基因敲除’小鼠来研究NG2-胶质细胞在这些功能中的重要性是困难的。为了克服这一点,我们的目标是确定NG2-胶质细胞是否可以使用免疫毒素方法进行靶向。我们证明,在体外,一次抗NG2抗体与二次皂苷偶联抗体共同孵育,选择性地杀死NG2表达细胞。此外,我们提供的证据表明,相同的方案导致NG2-胶质细胞的丢失,而不影响出生后小鼠小脑切片中星形胶质细胞或神经元的数量。本研究表明,免疫毒素靶向NG2-CSPG是一种有效的选择性杀伤NG2-胶质细胞的方法,对研究NG2-神经胶质细胞在正常中枢神经系统以及脱髓鞘和变性过程中的功能具有重要意义。
Expression of the transmembrane NG2 chondroitin sulphate proteoglycan (CSPG) defines a distinct population of NG2-glia. NG2-glia serve as a regenerative pool of oligodendrocyte progenitor cells in the adult central nervous system (CNS), which is important for demyelinating diseases such as multiple sclerosis, and are a major component of the glial scar that inhibits axon regeneration after CNS injury. In addition, NG2-glia form unique neuron–glial synapses with unresolved functions. However, to date it has proven difficult to study the importance of NG2-glia in any of these functions using conventional transgenic NG2 ‘knockout' mice. To overcome this, we aimed to determine whether NG2-glia can be targeted using an immunotoxin approach. We demonstrate that incubation in primary anti-NG2 antibody in combination with secondary saporin-conjugated antibody selectively kills NG2-expressing cells in vitro. In addition, we provide evidence that the same protocol induces the loss of NG2-glia without affecting astrocyte or neuronal numbers in cerebellar brain slices from postnatal mice. This study shows that targeting the NG2 CSPG with immunotoxins is an effective and selective means for killing NG2-glia, which has important implications for studying the functions of these enigmatic cells both in the normal CNS, and in demyelination and degeneration.
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