Cahoy John D, Emery Ben, Kaushal Amit, Foo Lynette C, Zamanian Jennifer L, Christopherson Karen S, Xing Yi, Lubischer Jane L, Krieg Paul A, Krupenko Sergey A, Thompson Wesley J, Barres Ben A
Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305, USA.
J Neurosci. 2008 Jan 2;28(1):264-78. doi: 10.1523/JNEUROSCI.4178-07.2008.
Understanding the cell-cell interactions that control CNS development and function has long been limited by the lack of methods to cleanly separate neural cell types. Here we describe methods for the prospective isolation and purification of astrocytes, neurons, and oligodendrocytes from developing and mature mouse forebrain. We used FACS (fluorescent-activated cell sorting) to isolate astrocytes from transgenic mice that express enhanced green fluorescent protein (EGFP) under the control of an S100beta promoter. Using Affymetrix GeneChip Arrays, we then created a transcriptome database of the expression levels of >20,000 genes by gene profiling these three main CNS neural cell types at various postnatal ages between postnatal day 1 (P1) and P30. This database provides a detailed global characterization and comparison of the genes expressed by acutely isolated astrocytes, neurons, and oligodendrocytes. We found that Aldh1L1 is a highly specific antigenic marker for astrocytes with a substantially broader pattern of astrocyte expression than the traditional astrocyte marker GFAP. Astrocytes were enriched in specific metabolic and lipid synthetic pathways, as well as the draper/Megf10 and Mertk/integrin alpha(v)beta5 phagocytic pathways suggesting that astrocytes are professional phagocytes. Our findings call into question the concept of a "glial" cell class as the gene profiles of astrocytes and oligodendrocytes are as dissimilar to each other as they are to neurons. This transcriptome database of acutely isolated purified astrocytes, neurons, and oligodendrocytes provides a resource to the neuroscience community by providing improved cell-type-specific markers and for better understanding of neural development, function, and disease.
长期以来，由于缺乏有效分离神经细胞类型的方法，对控制中枢神经系统（CNS）发育和功能的细胞间相互作用的理解一直受到限制。在此，我们描述了从发育中和成熟的小鼠前脑前瞻性分离和纯化星形胶质细胞、神经元和少突胶质细胞的方法。我们使用荧光激活细胞分选（FACS）从在S100β启动子控制下表达增强型绿色荧光蛋白（EGFP）的转基因小鼠中分离星形胶质细胞。然后，我们使用Affymetrix基因芯片阵列，通过对出生后第1天（P1）至P30不同出生后年龄的这三种主要CNS神经细胞类型进行基因谱分析，创建了一个包含>20,000个基因表达水平的转录组数据库。该数据库提供了对急性分离的星形胶质细胞、神经元和少突胶质细胞所表达基因的详细全面表征和比较。我们发现醛脱氢酶1L1（Aldh1L1）是星形胶质细胞的一种高度特异性抗原标志物，其星形胶质细胞表达模式比传统的星形胶质细胞标志物胶质纤维酸性蛋白（GFAP）广泛得多。星形胶质细胞在特定的代谢和脂质合成途径以及Draper/Megf10和Mertk/整合素α(v)β5吞噬途径中富集，这表明星形胶质细胞是专职吞噬细胞。我们的发现对“胶质”细胞类别的概念提出了质疑，因为星形胶质细胞和少突胶质细胞的基因谱彼此之间以及与神经元的基因谱一样不同。这个急性分离纯化的星形胶质细胞、神经元和少突胶质细胞的转录组数据库通过提供改进的细胞类型特异性标志物以及更好地理解神经发育、功能和疾病，为神经科学界提供了一种资源。
