Proteomic analysis of astrocytic secretion that regulates neurogenesis using quantitative amine-specific isobaric tagging.
Proteomic analysis of astrocytic secretion that regulates neurogenesis using quantitative amine-specific isobaric tagging.
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DOI：
10.1016/j.bbrc.2009.12.015
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发表时间：
2010-01
影响因子：
3.1
通讯作者：
Hu Yan;Wenhao Zhou;Liming Wei;F. Zhong;Yi Yang
中科院分区：
文献类型：
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作者：
Hu Yan;Wenhao Zhou;Liming Wei;F. Zhong;Yi Yang
星形胶质细胞是神经源性生态位的重要组成部分，通过膜结合和/或释放可溶性因子来影响神经发生。为了确定星形胶质细胞释放的调节神经干细胞分化和增殖的因子，我们使用轻度缺氧-葡萄糖剥夺(OGD)来抑制星形胶质细胞的分泌能力。使用Transwell共培养系统，我们发现OGD处理的星形胶质细胞不能促进神经干细胞的分化和增殖。接下来，采用等压标记相对和绝对定量(ITRAQ)蛋白质组学技术来鉴定星形胶质细胞(有或没有OGD)上清液中的蛋白质。通过多步骤分析和基因本体分类，共鉴定出130个胞外蛋白，其中大部分与神经元发育、炎症反应、细胞外基质组成和支持功能有关。在这些蛋白质中，有44种从未被报道是由星形胶质细胞产生的。使用ProteinPilot软件分析，我们发现OGD处理的星形胶质细胞上清液中有60个胞外蛋白发生显著变化(27个上调，33个下调)。在这些蛋白质中，已有7个蛋白质被报道能够调节神经发生，而其他蛋白质可能具有调节神经发生的潜力。这项研究描述了星形胶质细胞释放的主要蛋白质，这些蛋白质在神经发生的调节中发挥着重要作用。
Astrocytes are essential components of neurogenic niches that affect neurogenesis through membrane association and/or the release of soluble factors. To identify factors released from astrocytes that could regulate neural stem cell differentiation and proliferation, we used mild oxygen–glucose deprivation (OGD) to inhibit the secretory capacity of astrocytes. Using the Transwell co-culture system, we found that OGD-treated astrocytes could not promote neural stem cell differentiation and proliferation. Next, isobaric tagging for the relative and absolute quantitation (iTRAQ) proteomics techniques was performed to identify the proteins in the supernatants of astrocytes (with or without OGD). Through a multi-step analysis and gene ontology classification, 130 extracellular proteins were identified, most of which were involved in neuronal development, the inflammatory response, extracellular matrix composition and supportive functions. Of these proteins, 44 had never been reported to be produced by astrocytes. Using ProteinPilot software analysis, we found that 60 extracellular proteins were significantly altered (27 upregulated and 33 downregulated) in the supernatant of OGD-treated astrocytes. Among these proteins, 7 have been reported to be able to regulate neurogenesis, while others may have the potential to regulate neurogenesis. This study profiles the major proteins released by astrocytes, which play important roles in the modulation of neurogenesis.