Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut, USA.
Dev Neurobiol. 2010 Jan;70(1):1-15. doi: 10.1002/dneu.20750.
Many of the mitoses that produce pyramidal neurons in neocortex occur at the dorsolateral surface of the lateral ventricles in the embryo. RanBPM was found in a yeast two-hybrid screen to potentially interact with citron kinase (CITK), a protein shown previously to localize to the surface of the lateral ventricles and to be essential to neurogenic mitoses. Similar to its localization in epithelia, RanBPM protein is concentrated at the adherens junctions in developing neocortex. The biochemical interaction between CITK and RanBPM was confirmed in coimmunoprecipitation and protein overlay experiments. To test for a functional role of RanPBM in vivo, we used in utero RNAi. RanBPM RNAi decreased the polarization of CITK to the ventricular surface, increased the number of cells in mitosis, and decreased the number of cells in cytokinesis. Finally, the effect of RanBPM knockdown on mitosis was reversed in embryos mutant for CITK. Together, these results indicate that RanBPM, potentially through interaction with CITK, plays a role in the progression of neocortical precursors through M-phase at the ventricular surface.
许多产生新皮层锥体神经元的有丝分裂发生在胚胎侧脑室的背外侧表面。RanBPM 在酵母双杂交筛选中被发现可能与 citron 激酶 (CITK) 相互作用,CITK 先前被证明定位于侧脑室表面,对于神经发生有丝分裂至关重要。与上皮细胞中的定位相似,RanBPM 蛋白在发育中的新皮层中集中在黏着连接处。CITK 和 RanBPM 之间的生化相互作用在共免疫沉淀和蛋白覆盖实验中得到了证实。为了在体内测试 RanPBM 的功能作用,我们使用了子宫内 RNAi。RanBPM RNAi 降低了 CITK 向脑室表面的极化,增加了有丝分裂细胞的数量,并减少了细胞在胞质分裂中的数量。最后,RanBPM 敲低对有丝分裂的影响在 CITK 突变体胚胎中得到逆转。总之,这些结果表明,RanBPM 可能通过与 CITK 的相互作用,在新皮层前体通过 M 期在脑室表面的进展中发挥作用。