Identification of Proliferating and Migrating Cells by BrdU and Other Thymidine Analogs: Benefits and Limitations
Identification of Proliferating and Migrating Cells by BrdU and Other Thymidine Analogs: Benefits and Limitations
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DOI：
10.1007/978-1-4939-2313-7_7
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发表时间：
2015-01-01
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影响因子：
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通讯作者：
Rakic, Pasko
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文献类型：
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作者：
Duque, Alvaro;Rakic, Pasko
氚化胸苷 ([H-3]dT) 及其类似物溴脱氧尿苷 (BrdU)、碘脱氧尿苷 (IdU)、氯脱氧尿苷 (CldU) 和乙炔基脱氧尿苷 (EdU) 被广泛用于研究神经元的起源时间和迁移模式。在这里，我们讨论使用这些标记识别分裂细胞的优点和缺点，并强调它们只是指示 DNA 合成。因此，除了分裂细胞外，它们还可以标记正在进行 DNA 修复和/或细胞死亡的细胞。作为 DNA 中的外来分子，这些标记物可以影响细胞增殖的动力学，并且还具有不可预测的功能后果。我们回顾了一般方案，并根据我们自己对非人类灵长类动物的研究提供了证据，表明细胞增殖（如细胞数量和/或其存活率以及迁移细胞的定位所示）可能受到标记方法本身的影响。我们表明，使用 [H-3] dT 标记进行的测量比使用 BrdU 进行的测量更准确，这可能是因为 [H-3] dT 的毒性较小。因此，在解释使用胸苷类似物获得的结果作为有丝分裂数量、迁移模式、最终位置和细胞最终命运的可能指标时，应格外小心。
Tritiated thymidine ([H-3]dT) and its analogs bromodeoxyuridine (BrdU), iododeoxyuridine (IdU), chlorodeoxyuridine (CldU), and ethynyl deoxyuridine (EdU) are used extensively to study neuronal time of origin and pattern of migration. Here, we discuss the advantages and pitfalls of identifying dividing cells with these markers and emphasize that they simply indicate DNA synthesis. Thus, in addition to dividing cells, they can label cells that are undergoing DNA repair and/or cell death. As foreign molecules in the DNA, these markers can affect the kinetics of cell proliferation and also have unpredictable functional consequences. We review general protocols and present evidence, based on our own studies in nonhuman primates, that cell proliferation, as indicated by cell numbers and/or their survival, as well as the localization of migrating cells, may be affected by the labeling method itself. We show that measurements made using [H-3] dT labeling are more accurate than those using BrdU, perhaps because [H-3] dT is less toxic. Thus, utmost caution should be exercised when interpreting the results obtained by using thymidine analogs as possible indicators of the number of mitotic divisions, patterns of migration, final positions, and ultimate fate of cells.