LoVerso Peter R, Wachter Christopher M, Cui Feng
Thomas H. Gosnell School of Life Sciences, Rochester Institute of Technology, Rochester, NY, USA.
Bioinform Biol Insights. 2015 Nov 25;9:153-64. doi: 10.4137/BBI.S33124. eCollection 2015.
The mammalian brain is characterized by distinct classes of cells that differ in morphology, structure, signaling, and function. Dysregulation of gene expression in these cell populations leads to various neurological disorders. Neural cells often need to be acutely purified from animal brains for research, which requires complicated procedure and specific expertise. Primary culture of these cells in vitro is a viable alternative, but the differences in gene expression of cells grown in vitro and in vivo remain unclear. Here, we cultured three major neural cell classes of rat brain (ie, neurons, astrocytes, and oligodendrocyte precursor cells [OPCs]) obtained from commercial sources. We measured transcript abundance of these cell types by RNA sequencing (RNA-seq) and compared with their counterparts acutely purified from mouse brains. Cross-species RNA-seq data analysis revealed hundreds of genes that are differentially expressed between the cultured and acutely purified cells. Astrocytes have more such genes compared to neurons and OPCs, indicating that signaling pathways are greatly perturbed in cultured astrocytes. This dataset provides a powerful resource to demonstrate the similarities and differences of biological processes in mammalian neural cells grown in vitro and in vivo at the molecular level.
哺乳动物的大脑由形态、结构、信号传导和功能各异的不同类型细胞所构成。这些细胞群体中基因表达的失调会导致各种神经疾病。为了进行研究，常常需要从动物大脑中急性纯化神经细胞，这需要复杂的程序和专业技能。在体外对这些细胞进行原代培养是一种可行的替代方法，但体外培养的细胞与体内细胞在基因表达上的差异仍不明确。在这里，我们培养了从商业来源获得的大鼠大脑中的三种主要神经细胞类型（即神经元、星形胶质细胞和少突胶质前体细胞 [OPC]）。我们通过RNA测序（RNA-seq）测量了这些细胞类型的转录本丰度，并与从小鼠大脑中急性纯化得到的相应细胞进行了比较。跨物种RNA-seq数据分析揭示了数百个在培养细胞和急性纯化细胞之间差异表达的基因。与神经元和OPC相比，星形胶质细胞有更多这样的基因，这表明培养的星形胶质细胞中的信号通路受到了极大的干扰。该数据集为在分子水平上展示体外培养和体内生长的哺乳动物神经细胞生物学过程的异同提供了强大的资源。
