USE OF BROMODEOXYURIDINE-IMMUNOHISTOCHEMISTRY TO EXAMINE THE PROLIFERATION, MIGRATION AND TIME OF ORIGIN OF CELLS IN THE CENTRAL NERVOUS-SYSTEM
USE OF BROMODEOXYURIDINE-IMMUNOHISTOCHEMISTRY TO EXAMINE THE PROLIFERATION, MIGRATION AND TIME OF ORIGIN OF CELLS IN THE CENTRAL NERVOUS-SYSTEM
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DOI：
10.1016/0006-8993(88)90055-8
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发表时间：
1988-08-02
期刊：
影响因子：
2.9
通讯作者：
NOWAKOWSKI, RS
中科院分区：
文献类型：
--
作者：
MILLER, MW;NOWAKOWSKI, RS
本文用外源性胸苷类似物5-溴-2“-脱氧尿苷（BrdU）研究大脑皮层细胞的增殖、迁移和起源时间，并与[~ 3 H]胸苷（[~ 3 H]dT）放射自显影进行比较。给妊娠大鼠或小鼠注射BrdU和/或[3 H]dT，并通过标准免疫组织化学技术（使用单链DNA中针对BrdU的一抗）、放射自显影方法或两者进行处理。在注射后仅存活1小时的动物中，BrdU阳性细胞分布在整个中枢神经系统（CNS）的增殖区。在注射BrdU后1-3天处死的动物中，强烈的免疫反应性细胞位于表层皮质板中，而不太强烈的标记细胞分散在深层皮质板、中间区和生发区。在成年动物中，GD 19注射BrdU后60天或更长时间，BrdU阳性细胞位于新皮质的II/III层、海马锥体层和齿状回的颗粒层。在双标记研究中，BrdU免疫反应细胞的分布与放射自显影标记细胞的分布相同，并且所有放射自显影标记的神经元均为BrdU阳性。因此，BrdU免疫组化是适合的发育研究的中枢神经系统，此外，它提供了几个优势，[3 H]dT放射自显影。
The use of an exogenously administered thymidine analog, 5-bromo-2''-deoxyuridine (BrdU), for studies of the proliferation, migration and time of origin of cells in the cerebral cortex was investigated and compared with [3H]thymidine ([3H]dT) autoradiography. Pregnant rats or mice were injected with BrdU and/or [3H]dT and processed by standard immunohistochemical techniques using a primary antibody directed against BrdU in single-stranded DNA, autoradiographic methods, or both. In animals that survived only 1 h after the injection, BrdU-positive cells were distributed in the proliferative zones throughout the central nervous system (CNS). In animals killed 1-3 days after the BrdU injection, intensely immunoreactive cells were in the superficial cortical plate and less intensely labeled cells were scattered throughout the deep cortical plate, the intermediate zone, and the germinal zones. In adult animals, 60 days or more after an injection of BrdU on GD 19, BrdU-positive cells were located in layer II/III of neocortex, the hippocampal pyramidal layer, and the granule layer of the dentate gyrus. In the double-labeling studies, the distribution of BrdU-immunoreactive cells was identical to that of autoradiographically labeled cells, and all autoradiographically labeled neurons were BrdU positive. Thus, BrdU immunohistochemistry is suitable for developmental studies of the CNS; moreover, it provides several advantages over [3H]dT autoradiography.