Temporal mapping of gene expression levels during the differentiation of individual primary hematopoietic cells
Temporal mapping of gene expression levels during the differentiation of individual primary hematopoietic cells
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DOI：
10.1073/pnas.93.23.13158
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发表时间：
1996-11-12
影响因子：
11.1
通讯作者：
Scadden, DT
中科院分区：
文献类型：
--
作者：
Cheng, T;Shen, HM;Scadden, DT
已经提出了一种基因表达的分级顺序来控制造血中的发育事件，但直接证明调控基因表达和分化之间的时间关系一直很难实现。我们对单细胞聚合酶链式反应方法进行了改进，以检测每个细胞的mRNA拷贝数的2倍变化(动态范围为250-250,000拷贝/细胞)，并使用它来顺序定量单个原始(CD34(+)，CD38(-))祖细胞分化为红细胞、粒细胞或单核/巨噬细胞时的基因表达水平。检测分化标志物CD34或细胞因子受体mRNAs及与其调控相关的转录因子，所有被测转录因子均在多能祖细胞中表达。然而，在特定的谱系分化过程中，出现了不同的表达模式，在红系分化过程中，SCL、GATA-2和GATA-1的表达依次消失。PU.1、AML1B和C/EBPα在成熟粒细胞中的表达谱及其与细胞因子受体表达的关系可以与单核细胞中类似的谱相区别，这些数据表征了伴随血细胞发育的基因表达的动态，并定义了特定造血分化阶段的标志性基因表达模式。
A hierarchical order of gene expression has been proposed to control developmental events in hematopoiesis, but direct demonstration of the temporal relationships between regulatory gene expression and differentiation has been difficult to achieve. We modified a single-cell PCR method to detect 2-fold changes in mRNA copies per cell (dynamic range, 250-250,000 copies/cell) and used it to sequentially quantitate gene expression levels as single primitive (CD34(+),CD38(-)) progenitor cells underwent differentiation to become erythrocytes, granulocytes, or monocyte/macrophages. Markers of differentiation such as CD34 or cytokine receptor mRNAs and transcription factors associated with their regulation were assessed, All transcription factors tested were expressed in multipotent progenitors. During lineage-specific differentiation, however, distinct patterns of expression emerged, SCL, GATA-2, and GATA-1 expression sequentially extinguished during erythroid differentiation. PU.1, AML1B, and C/EBP alpha expression profiles and their relationship to cytokine receptor expression in maturing granulocytes could be distinguished from similar profiles in monocytic cells, These data characterize the dynamics of gene expression accompanying blood cell development and define a signature gene expression pattern for specific stages of hematopoietic differentiation.