Lewandowski Thomas A, Ponce Rafael A, Charleston Jay S, Hong Sungwoo, Faustman Elaine M
Department of Environmental Health, University of Washington, Seattle, WA 98105, USA.
Brain Res Dev Brain Res. 2003 Mar 14;141(1-2):117-28. doi: 10.1016/s0165-3806(03)00004-x.
We employed 5-bromo-2'-deoxyuridine (BrdU) labeling to identify in vivo changes in the cell cycle patterns of the rat midbrain during the major period of midbrain organogenesis, gestational days (gd) 11 to 16. We also used quantitative stereology to determine changes in absolute cell numbers during these gestational time points. Between gd 12 and 16, the length of S-phase did not change significantly while the fraction of cycling cells decreased from 73 to 11%. The average cell cycle length was determined to be 15 h on gd 12 and 17 h on gd 16, the difference not being statistically significant. The cell number in the midbrain increased from 1.3E5 cells on gd 11 to 1.7E7 cells on gd 16. On gd 12 and gd 13, there was a significant negative correlation between litter position and midbrain cell number, the effect diminishing on later days of gestation. The combined use of quantitative stereology and flow cytometry to study brain development represents a novel application that allows for simultaneous evaluation of changes in cell proliferation kinetics and the resulting effect of those kinetic changes on embryonic midbrain development.
我们采用5-溴-2'-脱氧尿苷（BrdU）标记法，以确定在中脑器官发生的主要时期，即妊娠第11至16天，大鼠中脑细胞周期模式的体内变化。我们还使用定量体视学方法来确定这些妊娠时间点期间绝对细胞数目的变化。在妊娠第12至16天之间，S期长度没有显著变化，而循环细胞的比例从73%降至11%。测定的平均细胞周期长度在妊娠第12天为15小时，在妊娠第16天为17小时，差异无统计学意义。中脑的细胞数量从妊娠第11天的1.3×10⁵个细胞增加到妊娠第16天的1.7×10⁷个细胞。在妊娠第12天和第13天，窝内位置与中脑细胞数量之间存在显著的负相关，这种影响在妊娠后期逐渐减弱。定量体视学和流式细胞术联合用于研究大脑发育是一种新的应用，它能够同时评估细胞增殖动力学的变化以及这些动力学变化对胚胎中脑发育的最终影响。
