Gilad G M, Gilad V H, Dahl D
Department of Neuropathology, Harvard Medical School, Boston, MA.
Neurosci Lett. 1989 Jan 2;96(1):7-12. doi: 10.1016/0304-3940(89)90234-6.
The developmental expression of neurofilaments immunoreactivity was examined in frozen sections and in primary cultures of rat cerebellum by immunocytochemistry with a series of monoclonal antibodies and with a polyclonal antibody. In tissue sections immunocytochemical staining with all the antibodies used was observed in basket cells where adult-like appearance could be detected by 14 days of age and adult-level intensity was achieved by about 25 days. Granule cells remained unstained. Intense staining appeared in cerebellar white matter as early as 7 days after birth. In contrast, neurofilaments immunoreactivity was detected in cultured granule cells from 7-day-old cerebellum. Only polyclonal antibodies reacting with the highly conserved middle alpha-helical domain of the neurofilament subunits were reactive in culture. Staining could be detected in the nerve cell bodies from the first day after plating; thereafter staining intensity increased and was also distributed in neurite extensions. We conclude that unlike their counterparts in vivo cultured embryonic granule cells can express certain neurofilaments immunoreactivity.
利用一系列单克隆抗体和一种多克隆抗体,通过免疫细胞化学方法,在大鼠小脑的冰冻切片和原代培养物中检测了神经丝免疫反应性的发育表达。在组织切片中,所用的所有抗体进行免疫细胞化学染色时,在篮状细胞中均观察到染色,在14日龄时可检测到类似成年的外观,到约25日龄时达到成年水平强度。颗粒细胞未染色。出生后7天,小脑白质中就出现了强烈染色。相比之下,在7日龄小脑的培养颗粒细胞中检测到了神经丝免疫反应性。只有与神经丝亚基高度保守的中间α螺旋结构域反应的多克隆抗体在培养物中有反应性。接种后第一天就能在神经细胞体中检测到染色;此后染色强度增加,且也分布在神经突延伸部位。我们得出结论,与体内对应物不同,培养的胚胎颗粒细胞能够表达某些神经丝免疫反应性。