Weisheit Gunnar, Gliem Michael, Endl Elmar, Pfeffer Peter L, Busslinger Meinrad, Schilling Karl
Anatomisches Institut, Anatomie and Zellbiologie, University of Bonn, Bonn, Germany.
Eur J Neurosci. 2006 Jul;24(2):466-78. doi: 10.1111/j.1460-9568.2006.04915.x.
The cerebellar cortex consists of a small set of neuronal cell types interconnected in a highly stereotyped way. While the development of cerebellar cortical projection neurons, i.e. Purkinje cells, and that of granule cells has been elucidated in considerable detail, that of cerebellar cortical inhibitory interneurons is still rather fragmentarily understood. Here, we use mice expressing green fluorescent protein (GFP) from the Pax2 locus to analyse the ontogenesis of these cells. Numbers of Pax2-positive inhibitory interneuronal precursors increase following a classical sigmoidal growth curve to yield a total of some 905.000 +/- 77.000 cells. Maximal cell increase occurs at about postnatal day (P)5.4, and some 75% of all inhibitory interneurons are generated prior to P7. Conjoint analysis of the developmental accruement of Pax2-GFP-positive cells and their cell cycle distribution reveals that, at least at P0 and P3, the numerical increase of these cells results primarily from proliferation of a Pax2-negative precursor population and suggests that Pax2 expression begins at or around the final mitosis. Following their terminal mitosis, inhibitory cerebellar cortical interneurons go through a protracted quiescent phase in which they maintain expression of the cell cycle marker Ki-67. During this phase, they translocate into the nascent molecular layer, where they stall next to premigratory granule cell precursors without penetrating this population of cells. These observations provide a quantitative description of cerebellar cortical inhibitory interneuron genesis and early differentiation, and define Pax2 as a marker expressed in basket and stellate cells, from around their final mitosis to their incipient histogenetic integration.
小脑皮质由一小部分以高度固定方式相互连接的神经元细胞类型组成。虽然小脑皮质投射神经元(即浦肯野细胞)和颗粒细胞的发育已得到相当详细的阐明,但小脑皮质抑制性中间神经元的发育仍了解得相当零散。在这里,我们使用从Pax2基因座表达绿色荧光蛋白(GFP)的小鼠来分析这些细胞的个体发生。Pax2阳性抑制性中间神经元前体细胞的数量遵循经典的S形生长曲线增加,最终产生约905,000±77,000个细胞。最大细胞增加发生在出生后约第5.4天,并且所有抑制性中间神经元中约75%在出生后第7天之前产生。对Pax2-GFP阳性细胞的发育积累及其细胞周期分布的联合分析表明,至少在出生后第0天和第3天,这些细胞的数量增加主要源于Pax2阴性前体细胞群的增殖,这表明Pax2表达在最终有丝分裂时或前后开始。在终末有丝分裂后,小脑皮质抑制性中间神经元经历一个漫长的静止期,在此期间它们维持细胞周期标记物Ki-67的表达。在此阶段,它们迁移到新生分子层,在那里它们停留在迁移前颗粒细胞前体旁边,而不穿透这群细胞。这些观察结果提供了小脑皮质抑制性中间神经元发生和早期分化的定量描述,并将Pax2定义为在篮状细胞和星状细胞中从最终有丝分裂前后到其初始组织发生整合期间表达的标记物。