Unidad de Citología e Histología, Departament de Biologia Cellular, de Fisiologia i d'Immunologia, Facultad de Biociencias, Institut de Neurociències, Universidad Autónoma de Barcelona , Barcelona, Spain.
Cell Cycle. 2020 Oct;19(19):2451-2459. doi: 10.1080/15384101.2020.1806425. Epub 2020 Aug 23.
Cerebellar neurons are generated from the rhombic lip and the neuroepithelium. In this study, we analyze the histogenesis of the cerebellar neuroepithelium in terms of cellular kinetics. The experimental animals are the offspring of pregnant dams injected with 5-bromo-2'-deoxyuridine (BrdU) on embryonic day 13. We infer the fraction of S-phase cells by examining a range of survival times after a single BrdU-exposure and a cumulative BrdU-labeling sequence, which allow for the derivation of cell-cycle parameters and phase durations. The current results indicate that the dose of BrdU employed (35 mg/kg) provides saturation S-phase labeling from at least 1 h after marker delivery. The duration of G2, mitotic phase, and G1 are 1.2, 0.5, and 6.9 h, respectively. The duration for the S-phase, growth fraction, and the whole cycle are obtained on the basis of two proliferative models, steady-state and exponential growth. Both models provided similar results. In conclusion, our results indicate that the steady-state and the cumulative S-phase labeling paradigms can be adopted to analyze cell cycle parameters in the cerebellar neuroepithelium. Current results can help in understanding the regulatory mechanisms of cerebellar histogenesis and the cell biological mechanisms of the proliferative cycle of the neuroepithelium.
小脑神经元由菱唇和神经上皮产生。在这项研究中,我们从细胞动力学的角度分析了小脑神经上皮的组织发生。实验动物是在胚胎第 13 天给怀孕的母鼠注射 5-溴-2'-脱氧尿苷(BrdU)的后代。我们通过检查单次 BrdU 暴露后的一系列存活时间和累积 BrdU 标记序列来推断 S 期细胞的分数,这允许推导细胞周期参数和相持续时间。目前的结果表明,所使用的 BrdU 剂量(35mg/kg)在标记物传递后至少 1 小时提供饱和的 S 期标记。G2、有丝分裂期和 G1 的持续时间分别为 1.2、0.5 和 6.9 小时。基于稳态和指数生长两种增殖模型,获得了 S 期、生长分数和整个周期的持续时间。两种模型都提供了类似的结果。总之,我们的结果表明,稳态和累积 S 期标记范式可用于分析小脑神经上皮的细胞周期参数。目前的结果有助于理解小脑组织发生的调控机制和神经上皮增殖周期的细胞生物学机制。