Massachusetts Eye and Ear, Boston, MA.
Zuckerman Mind Brain Behavior Institute, Columbia University, New York, NY.
Mol Biol Cell. 2024 Oct 1;35(10):ar129. doi: 10.1091/mbc.E24-05-0217. Epub 2024 Aug 21.
Nde1 is a cytoplasmic dynein regulatory protein with important roles in vertebrate brain development. One noteworthy function is in the nuclear oscillatory behavior in neural progenitor cells, the control and mechanism of which remain poorly understood. Nde1 contains multiple phosphorylation sites for the cell cycle-dependent protein kinase CDK1, though the function of these sites is not well understood. To test their role in brain development, we expressed phosphorylation-state mutant forms of Nde1 in embryonic rat brains using in utero electroporation. We find that Nde1 T215 and T243 phosphomutants block apical interkinetic nuclear migration (INM) and, consequently, mitosis in radial glial progenitor cells. Another Nde1 phosphomutant at T246 also interfered with mitotic entry without affecting INM, suggesting a more direct role for Nde1 T246 in mitotic regulation. We also found that the Nde1 S214F mutation, which is associated with schizophrenia, inhibits Cdk5 phosphorylation at an adjacent residue which causes alterations in neuronal lamination. These results together identify important new roles for Nde1 phosphorylation in neocortical development and disease, and represent the first evidence for Nde1 phosphorylation roles in INM and neuronal lamination.
Nde1 是一种细胞质动力蛋白调节蛋白,在脊椎动物大脑发育中具有重要作用。其一个显著功能是在神经祖细胞的核振荡行为中,但其调控和机制仍知之甚少。Nde1 包含多个细胞周期依赖性蛋白激酶 CDK1 的磷酸化位点,尽管这些位点的功能尚不清楚。为了测试它们在大脑发育中的作用,我们使用子宫内电穿孔在胚胎大鼠脑中表达了 Nde1 的磷酸化状态突变体。我们发现 Nde1 T215 和 T243 磷酸化突变体阻止了顶端细胞有丝分裂核迁移(INM),从而阻止了放射状神经胶质祖细胞的有丝分裂。另一种 Nde1 磷酸化突变体 T246 也干扰了有丝分裂的进入,而不影响 INM,这表明 Nde1 T246 在有丝分裂调控中具有更直接的作用。我们还发现,与精神分裂症相关的 Nde1 S214F 突变抑制了 Cdk5 在相邻残基上的磷酸化,从而导致神经元分层的改变。这些结果共同确定了 Nde1 磷酸化在新皮层发育和疾病中的重要新作用,并且代表了 Nde1 在 INM 和神经元分层中的磷酸化作用的第一个证据。