Synthetic Thymidine Analog Labeling without Misconceptions.

Synthetic Thymidine Analog Labeling without Misconceptions.
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DOI:
10.3390/cells11121888
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发表时间:
2022-06-10
期刊:
影响因子:
6
通讯作者:
--
中科院分区:
生物学2区
文献类型:
--
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用胸苷类似物标记增殖细胞是不可或缺的研究工具;然而,这些化合物潜在的体内细胞毒性问题仍未解决。在这里,我们通过检查 BrdU 和 EdU 对成年海马神经发生的影响以及 EdU 对小鼠围产期体细胞发育的影响来解决这些问题。我们发现,在各种剂量下,EdU 和 BrdU 在给药后不久就会在齿状回中标记相似数量的细胞。此外,虽然注射 EdU 不会影响注射后 48 小时内神经祖细胞的分裂和存活,但根据 6 周后的评估,它确实会影响细胞存活。我们还表明,在出生后第一天单次注射不同剂量的 EdU 不会导致第一周内一组形态测量标准发生明显变化;然而,较高剂量的 EdU 会对幼鼠随后的体细胞成熟和大脑生长产生不利影响。我们的结果表明使用胸苷类似物标记复制 DNA 时可能需要注意的事项,并提出了应用该方法的指南。
Tagging proliferating cells with thymidine analogs is an indispensable research tool; however, the issue of the potential in vivo cytotoxicity of these compounds remains unresolved. Here, we address these concerns by examining the effects of BrdU and EdU on adult hippocampal neurogenesis and EdU on the perinatal somatic development of mice. We show that, in a wide range of doses, EdU and BrdU label similar numbers of cells in the dentate gyrus shortly after administration. Furthermore, whereas the administration of EdU does not affect the division and survival of neural progenitor within 48 h after injection, it does affect cell survival, as evaluated 6 weeks later. We also show that a single injection of various doses of EdU on the first postnatal day does not lead to noticeable changes in a panel of morphometric criteria within the first week; however, higher doses of EdU adversely affect the subsequent somatic maturation and brain growth of the mouse pups. Our results indicate the potential caveats in labeling the replicating DNA using thymidine analogs and suggest guidelines for applying this approach.
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