Menezes J R, Luskin M B
Department of Anatomy and Cell Biology, Emory University School of Medicine, Atlanta, Georgia 30322.
J Neurosci. 1994 Sep;14(9):5399-416. doi: 10.1523/JNEUROSCI.14-09-05399.1994.
We have used a monoclonal antibody against the neuron-specific class III beta-tubulin (TuJ1; Lee et al., 1990b) to study the distribution and morphology of immature neurons in the proliferative ventricular and subventricular zones of the developing telencephalon. Mouse brains from embryonic day 12 (E12) to postnatal day 5 (P5) were fixed either with a non-cross-linking agent, HistoChoice, or with 4% paraformaldehyde, and processed for TuJ1 immunohistochemistry. TuJ1 immunoreactivity first appeared in the proliferative zones of the developing cerebral cortex at E13-E14 as the cortical plate was emerging. After E14, tangentially oriented TuJ1-positive cells were abundant at the interface between the ventricular and subventricular zones. This tangential pattern was less conspicuous in the developing striatum. Within the cortical and striatal ventricular zone TuJ1-positive cells were less numerous and displayed a variety of orientations and morphologies. Postnatally, after the period of neurogenesis has ended, TuJ1 immunoreactivity continued to increase in the subventricular zone and remained high until the last developmental stage examined (P5). Anti-MAP2, another neuron-specific marker, never labeled the cells of the ventricular and subventricular zones, pre- or postnatally. To determine the birthdates of TuJ1-positive cells in the cortical-ventricular and subventricular zones, brains were double labeled with TuJ1 and bromodeoxyuridine according to different pulse-chase schedules. TuJ1-positive cells were postmitotic and generated throughout the period of cortical neurogenesis. Collectively, the results suggest that TuJ1 immunoreactivity distinguishes two neuronal populations: those that remain for an indefinite period of time in the proliferative zones, and those that leave the proliferative zones soon after being generated. Although the fate of the TuJ1-positive cells that reside in the proliferative zones remains unclear, their tangentially aligned orientation and their distribution suggest that they migrate independent of radial glial fibers.
我们运用了一种针对神经元特异性Ⅲ类β-微管蛋白(TuJ1;Lee等人,1990b)的单克隆抗体,来研究发育中的端脑增殖性脑室区和脑室下区中未成熟神经元的分布及形态。取自胚胎第12天(E12)至出生后第5天(P5)的小鼠脑,用非交联剂HistoChoice或4%多聚甲醛固定,然后进行TuJ1免疫组织化学处理。随着皮质板的出现,TuJ1免疫反应性在E13 - E14时首次出现在发育中的大脑皮质增殖区。E14之后,在脑室区和脑室下区的界面处,沿切线方向排列的TuJ1阳性细胞大量存在。这种切线模式在发育中的纹状体中不太明显。在皮质和纹状体脑室区内,TuJ1阳性细胞数量较少,呈现出各种取向和形态。出生后,在神经发生期结束后,TuJ1免疫反应性在脑室下区持续增加,并一直保持高水平,直到所检测的最后发育阶段(P5)。另一种神经元特异性标记物抗MAP2,在出生前或出生后都从未标记过脑室区和脑室下区的细胞。为了确定皮质脑室区和脑室下区中TuJ1阳性细胞的出生日期,根据不同的脉冲追踪方案,用TuJ1和溴脱氧尿苷对大脑进行双重标记。TuJ1阳性细胞已完成有丝分裂,并且在整个皮质神经发生期都有产生。总体而言,结果表明TuJ1免疫反应性区分了两类神经元群体:一类在增殖区停留不确定的时间,另一类在产生后不久就离开增殖区。尽管位于增殖区的TuJ1阳性细胞的命运仍不清楚,但它们沿切线排列的取向和分布表明,它们的迁移独立于放射状胶质纤维。