Department of Neurobiology, Stanford University School of Medicine, Stanford, California 94305-5125,
The Vivian L. Smith Department of Neurosurgery, University of Texas Medical School at Houston, Houston, Texas 77057, Center for Stem Cell and Regenerative Medicine, University of Texas Brown Institute of Molecular Medicine, Houston, Texas 77057.
J Neurosci. 2014 Sep 3;34(36):11929-47. doi: 10.1523/JNEUROSCI.1860-14.2014.
The major cell classes of the brain differ in their developmental processes, metabolism, signaling, and function. To better understand the functions and interactions of the cell types that comprise these classes, we acutely purified representative populations of neurons, astrocytes, oligodendrocyte precursor cells, newly formed oligodendrocytes, myelinating oligodendrocytes, microglia, endothelial cells, and pericytes from mouse cerebral cortex. We generated a transcriptome database for these eight cell types by RNA sequencing and used a sensitive algorithm to detect alternative splicing events in each cell type. Bioinformatic analyses identified thousands of new cell type-enriched genes and splicing isoforms that will provide novel markers for cell identification, tools for genetic manipulation, and insights into the biology of the brain. For example, our data provide clues as to how neurons and astrocytes differ in their ability to dynamically regulate glycolytic flux and lactate generation attributable to unique splicing of PKM2, the gene encoding the glycolytic enzyme pyruvate kinase. This dataset will provide a powerful new resource for understanding the development and function of the brain. To ensure the widespread distribution of these datasets, we have created a user-friendly website (http://web.stanford.edu/group/barres_lab/brain_rnaseq.html) that provides a platform for analyzing and comparing transciption and alternative splicing profiles for various cell classes in the brain.
大脑的主要细胞类型在其发育过程、代谢、信号转导和功能上存在差异。为了更好地理解构成这些细胞类型的细胞的功能和相互作用，我们从小鼠大脑皮层中急性纯化了具有代表性的神经元、星形胶质细胞、少突胶质前体细胞、新形成的少突胶质细胞、髓鞘形成的少突胶质细胞、小胶质细胞、内皮细胞和周细胞群体。我们通过 RNA 测序为这 8 种细胞类型生成了转录组数据库，并使用敏感的算法检测每种细胞类型中的选择性剪接事件。生物信息学分析鉴定了数千种新的细胞类型特异性基因和剪接异构体，它们将为细胞鉴定提供新的标记物、遗传操作的工具，并深入了解大脑的生物学。例如，我们的数据为神经元和星形胶质细胞在动态调节糖酵解通量和归因于 PKM2（编码糖酵解酶丙酮酸激酶的基因）独特剪接的乳酸生成能力方面的差异提供了线索。该数据集将为理解大脑的发育和功能提供一个强大的新资源。为确保这些数据集的广泛传播，我们创建了一个用户友好的网站（http://web.stanford.edu/group/barres_lab/brain_rnaseq.html），该网站提供了一个平台，用于分析和比较大脑中各种细胞类型的转录和选择性剪接谱。
