溴脱氧尿苷和[3H]胸腺嘧啶核苷掺入 DNA 对细胞增殖、位置和命运的不同影响。

Different effects of bromodeoxyuridine and [3H]thymidine incorporation into DNA on cell proliferation, position, and fate.

机构信息

Department of Neurobiology, Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

出版信息

J Neurosci. 2011 Oct 19;31(42):15205-17. doi: 10.1523/JNEUROSCI.3092-11.2011.

Abstract

As markers of DNA synthesis, [(3)H]thymidine ([(3)H]dT) and the later developed analog bromodeoxyuridine (BrdU) have revolutionized our ability to identify dividing cells and follow their fate in various tissues, including the nervous system. However, the effect of the incorporation of these molecules into DNA on cell proliferation, migration, differentiation, and function is not fully understood. Here, we compare the number and distribution of labeled cells in the cerebral cortex of postnatal macaque monkeys exposed to either [(3)H]dT or BrdU as embryos. The large size and prolonged brain development in this species allows higher resolution of cellular events and more accurate discrimination between the two methods. Our analysis revealed substantial differences in the number and distribution of labeled cells. The data indicate that random incorporation of the thymidine analog BrdU into the genes of dividing cells makes the fate of postmitotic neurons more prone to unpredictable errors than the incorporation of the more natural DNA constituent nucleotide [(3)H]dT. These findings have implications for the interpretation of results obtained by BrdU as an index of the number of neurons produced, and their migration, placement, subsequent connectivity, function, and survival.

摘要

作为 DNA 合成的标志物,[(3)H]胸腺嘧啶核苷([(3)H]dT)和后来开发的类似物溴脱氧尿苷(BrdU)极大地提高了我们识别分裂细胞并追踪其在包括神经系统在内的各种组织中的命运的能力。然而,这些分子掺入 DNA 对细胞增殖、迁移、分化和功能的影响尚未完全了解。在这里,我们比较了胚胎期暴露于[(3)H]dT 或 BrdU 的新生猕猴大脑皮层中标记细胞的数量和分布。该物种的大脑较大且发育时间较长,允许对细胞事件进行更高分辨率的分析,并且可以更准确地区分这两种方法。我们的分析揭示了标记细胞数量和分布的显著差异。这些数据表明,随机将胸腺嘧啶类似物 BrdU 掺入分裂细胞的基因中,使得有丝分裂后神经元的命运比掺入更自然的 DNA 组成部分核苷酸[(3)H]dT 更容易受到不可预测的错误影响。这些发现对将 BrdU 作为产生的神经元数量及其迁移、定位、随后的连接、功能和存活的指标的结果的解释具有重要意义。

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