Hauke C, Ackermann I, Korr H
Institute of Anatomy, RWTH Aachen, Germany.
Cell Prolif. 1995 Nov;28(11):595-607. doi: 10.1111/j.1365-2184.1995.tb00047.x.
Pulse labelling experiments with [3H] thymidine (dT) and double labelling experiments with [3H]dT and bromodeoxyuridine (BrdUrd) were carried out on cells of the subependymal layer in the brain of adult normal mice in vivo, in vivo/in vitro and in vitro. The results should (i) lead to information about cell cycle parameters of these cells in the brain of adult mice, since these cells have been studied mostly in the rat brain up to now and (ii) answer the question whether results concerning cell proliferation obtained in vivo correspond with those from brain slices incubated in vitro with or without prelabelling in vivo. In vivo an LI of 20.2 +/- 2.7% (mean +/- SEM) and Ts = 7.2 +/- 0.7 h were found. Furthermore, grain count halving experiments led to a surprisingly short cycle time (Tc) of 11.2-14.2 h. The longer Tc values (18-20 h) reported in the literature for subependymal cells in the rat brain seem to be due to evaluations of different areas around the lateral ventricle without considering the migrating behaviour of these cells which is quite different regionally. The in vitro studies (with or without prelabelling in vivo) showed a significantly reduced LI due to the fact that about 20% of the S phase cells, possibly lying in the middle of S, stopped further DNA synthesis after transfer to culture. This was shown by comparing the cell fluxes at the G1/S and S/G2 borders of in vivo vs. in vitro studies.
用[³H]胸腺嘧啶核苷(dT)进行脉冲标记实验,并用[³H]dT和溴脱氧尿苷(BrdUrd)进行双标记实验,分别在成年正常小鼠脑室内、体内/体外和体外条件下对室管膜下层细胞进行实验。这些结果应能:(i)提供成年小鼠脑内这些细胞的细胞周期参数信息,因为到目前为止这些细胞大多是在大鼠脑中进行研究的;(ii)回答体内获得的细胞增殖结果是否与体内预标记或未预标记的脑片在体外培养所得到的结果一致的问题。在体内实验中,发现标记指数(LI)为20.2±2.7%(平均值±标准误),S期时长(Ts)为7.2±0.7小时。此外,银粒计数减半实验得出的细胞周期时间(Tc)短得出奇,为11.2 - 14.2小时。文献中报道的大鼠脑室管膜下细胞的较长Tc值(18 - 20小时),似乎是由于对侧脑室周围不同区域进行评估时未考虑这些细胞在不同区域有很大差异的迁移行为。体外研究(无论体内是否预标记)显示标记指数显著降低,原因是约20%处于S期中间的细胞在转移到培养液后停止了进一步的DNA合成。通过比较体内和体外研究中G1/S和S/G2边界处的细胞通量得以证实。