Tran Phuong B, Ren Dongjun, Veldhouse Thomas J, Miller Richard J
Department of Molecular Pharmacology and Biological Chemistry, Feinberg School of Medicine, Northwestern University, Chicago, Illinois 60611, USA.
J Neurosci Res. 2004 Apr 1;76(1):20-34. doi: 10.1002/jnr.20001.
We investigated the expression and functions of chemokine receptors in neural progenitor cells isolated from embryonic and adult mice. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis demonstrated mRNA expression for most known chemokine receptors in neural progenitor cells grown as neurospheres from embryonic (E17) and adult (4-week-old) mice. The expression of CXCR4 receptors was demonstrated further in E17 neurospheres using immunohistochemistry, in situ hybridization, Northern blot analysis and fura-2-based Ca(2+) imaging. Most neurospheres grown from E17 mice responded to stromal cell-derived factor-1 (SDF-1/CXCL12) in Ca(2+) imaging studies. In addition, immunohistochemical studies demonstrated that these neurospheres consisted of dividing cells that uniformly colocalized nestin and CXCR4 receptors. Differentiation of E17 neurospheres yielded astrocytes and neurons exhibiting several different phenotypes, including expression of calbindin, calretinin, gamma-aminobutyric acid (GABA), and glutamate, and many also coexpressed CXCR4 receptors. In addition, neurospheres grown from the subventricular zone (SVZ) of 4-week-old mice exhibited large increases in Ca(2+) in response to CXCL12 and several other chemokines. In comparison, neurospheres prepared from olfactory bulb of adult mice exhibited only small Ca(2+) responses to CXCL12, whereas neurospheres prepared from hippocampus were insensitive to CXCL12, although they did respond to other chemokines. Investigations designed to investigate whether CXCL12 can act as a chemoattractant demonstrated that cells dissociated from E17 or adult SVZ neurospheres migrated toward an CXCL12 gradient and this was blocked by the CXCR4 antagonist AMD3100. These results illustrate widespread chemokine sensitivity of embryonic and adult neural progenitor cells and support the view that chemokines may be of general importance in control of progenitor cell migration in embryonic and adult brain.
我们研究了从胚胎和成年小鼠分离的神经祖细胞中趋化因子受体的表达及功能。逆转录聚合酶链反应(RT-PCR)分析表明,以胚胎(E17)和成年(4周龄)小鼠的神经球形式生长的神经祖细胞中,大多数已知趋化因子受体都有mRNA表达。利用免疫组织化学、原位杂交、Northern印迹分析和基于fura-2的Ca(2+)成像进一步证实了E17神经球中CXCR4受体的表达。在Ca(2+)成像研究中,大多数从E17小鼠生长的神经球对基质细胞衍生因子-1(SDF-1/CXCL12)有反应。此外,免疫组织化学研究表明,这些神经球由均匀共定位巢蛋白和CXCR4受体的分裂细胞组成。E17神经球的分化产生了星形胶质细胞和神经元,它们表现出几种不同的表型,包括钙结合蛋白、钙视网膜蛋白、γ-氨基丁酸(GABA)和谷氨酸的表达,许多细胞还共表达CXCR4受体。此外,从4周龄小鼠脑室下区(SVZ)生长的神经球对CXCL12和其他几种趋化因子的反应表现为Ca(2+)大幅增加。相比之下,成年小鼠嗅球制备的神经球对CXCL12仅表现出较小的Ca(2+)反应,而海马制备的神经球对CXCL12不敏感,尽管它们对其他趋化因子有反应。旨在研究CXCL12是否可作为趋化因子的研究表明,从E17或成年SVZ神经球解离的细胞向CXCL12梯度迁移,且这种迁移被CXCR4拮抗剂AMD3100阻断。这些结果说明了胚胎和成年神经祖细胞对趋化因子具有广泛的敏感性,并支持趋化因子可能在胚胎和成年大脑中祖细胞迁移控制中具有普遍重要性的观点。