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对表达NG2硫酸软骨素蛋白聚糖的细胞进行免疫消融。

Immunoablation of cells expressing the NG2 chondroitin sulphate proteoglycan.

作者信息

Leoni Giampaolo, Rattray Marcus, Fulton Daniel, Rivera Andrea, Butt Arthur M

机构信息

Institute of Biology and Biomedical Sciences, School of Pharmacy and Biomedical Sciences, University of Portsmouth, Portsmouth, UK.

出版信息

J Anat. 2014 Feb;224(2):216-27. doi: 10.1111/joa.12141. Epub 2013 Nov 20.

DOI:10.1111/joa.12141
PMID:24252088
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3969064/
Abstract

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.

摘要

跨膜神经胶质2硫酸软骨素蛋白聚糖(CSPG)的表达定义了一个独特的神经胶质2细胞群体。神经胶质2细胞在成体中枢神经系统(CNS)中作为少突胶质细胞祖细胞的再生池,这对诸如多发性硬化症等脱髓鞘疾病很重要,并且是胶质瘢痕的主要成分,该胶质瘢痕会抑制中枢神经系统损伤后轴突的再生。此外,神经胶质2细胞形成了功能尚未明确的独特的神经元 - 胶质细胞突触。然而,迄今为止,使用传统的转基因神经胶质2“敲除”小鼠来研究神经胶质2细胞在这些功能中的任何一项的重要性都已证明很困难。为了克服这一问题,我们旨在确定是否可以使用免疫毒素方法靶向神经胶质2细胞。我们证明,在原代抗神经胶质2抗体与二级结合皂草素的抗体结合下孵育可在体外选择性杀死表达神经胶质2的细胞。此外,我们提供证据表明,相同的方案可诱导神经胶质2细胞的缺失,而不影响出生后小鼠小脑脑片中星形胶质细胞或神经元的数量。这项研究表明,用免疫毒素靶向神经胶质2 CSPG是杀死神经胶质2细胞的一种有效且选择性的方法,这对于研究这些神秘细胞在正常中枢神经系统以及脱髓鞘和变性中的功能具有重要意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/9468bd00e621/joa0224-0216-f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/ebd98372394f/joa0224-0216-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/74ac416ffee1/joa0224-0216-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/bc6cfc4ce491/joa0224-0216-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/e12f098f7597/joa0224-0216-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/36997cc3f695/joa0224-0216-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/38187fb1eaa3/joa0224-0216-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/bbc88bc19f6d/joa0224-0216-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/9468bd00e621/joa0224-0216-f8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/ebd98372394f/joa0224-0216-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/74ac416ffee1/joa0224-0216-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/bc6cfc4ce491/joa0224-0216-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/e12f098f7597/joa0224-0216-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/36997cc3f695/joa0224-0216-f5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/38187fb1eaa3/joa0224-0216-f6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/bbc88bc19f6d/joa0224-0216-f7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b4e/3969064/9468bd00e621/joa0224-0216-f8.jpg

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本文引用的文献

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Postnatal switch from synaptic to extrasynaptic transmission between interneurons and NG2 cells.神经元和 NG2 细胞之间的突触后传递从突触后转换为 extrasynaptic 传递。
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The NG2 proteoglycan promotes oligodendrocyte progenitor proliferation and developmental myelination.NG2 蛋白聚糖促进少突胶质前体细胞增殖和发育性髓鞘形成。
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NG2 cells differentiate into astrocytes in cerebellar slices.NG2细胞在小脑切片中分化为星形胶质细胞。
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Axons and astrocytes release ATP and glutamate to evoke calcium signals in NG2-glia.轴突和星形胶质细胞释放 ATP 和谷氨酸,以在 NG2 胶质细胞中引发钙信号。
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PDGFRA/NG2 glia generate myelinating oligodendrocytes and piriform projection neurons in adult mice.血小板衍生生长因子受体A/神经胶质抗原2(PDGFRA/NG2)神经胶质细胞在成年小鼠中产生形成髓鞘的少突胶质细胞和梨状投射神经元。
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