Rpsa Signaling Regulates Cortical Neuronal Morphogenesis via Its Ligand, PEDF, and Plasma Membrane Interaction Partner, Itga6
Rpsa Signaling Regulates Cortical Neuronal Morphogenesis via Its Ligand, PEDF, and Plasma Membrane Interaction Partner, Itga6
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DOI：
10.1093/cercor/bhab242
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发表时间：
2021-08-04
期刊：
影响因子：
3.7
通讯作者：
Toyo-Oka, Kazuhito
中科院分区：
文献类型：
--
作者：
Blazejewski, Sara M.;Bennison, Sarah A.;Toyo-Oka, Kazuhito
神经形态学缺陷是神经发育障碍和功能缺陷的基础。我们确定了 Rpsa 在调节神经形态发生中的功能，使用子宫内电穿孔敲低 Rpsa，导致顶端树突定向错误、延伸更少/更短、脊柱密度降低、上神经元层的脊柱形态改变以及上/下皮质层的树枝化减少。 Rpsa 敲低会破坏皮质发育的多个方面，包括放射状胶质细胞纤维形态和神经元分层。我们研究了 Rpsa 的配体 PEDF 和质膜上的相互作用伙伴 Itga6。 Rpsa、PEDF 和 Itga6 敲低会导致相似的表型，其中 Rpsa 和 Itga6 过表达可挽救体内 PEDF 缺陷神经元的形态缺陷。此外，Itga6 过表达可增加并稳定质膜上的 Rpsa 表达。 GCaMP6s 用于通过离体钙成像对 Rpsa 敲低进行功能分析。 Rpsa 缺陷的神经元荧光强度波动较小，表明阈下钙信号传导存在缺陷。编码 PEDF 的 Serpinf1 基因位于染色体 17p13.3，该基因在神经发育障碍米勒-迪克综合征患者中被删除。我们的研究确定了 Rpsa 在早期皮质发育中的作用以及 PEDF-Rpsa-Itga6 信号在神经形态发生中的作用，从而表明这些分子与米勒-迪克综合征等神经发育障碍的病因学有关，并将它们确定为潜在的治疗方法。
Neuromorphological defects underlie neurodevelopmental disorders and functional defects. We identified a function for Rpsa in regulating neuromorphogenesis using in utero electroporation to knockdown Rpsa, resulting in apical dendrite misorientation, fewer/shorter extensions, and decreased spine density with altered spine morphology in upper neuronal layers and decreased arborization in upper/lower cortical layers. Rpsa knockdown disrupts multiple aspects of cortical development, including radial glial cell fiber morphology and neuronal layering. We investigated Rpsa's ligand, PEDF, and interacting partner on the plasma membrane, Itga6. Rpsa, PEDF, and Itga6 knockdown cause similar phenotypes, with Rpsa and Itga6 overexpression rescuing morphological defects in PEDF-deficient neurons in vivo. Additionally, Itga6 overexpression increases and stabilizes Rpsa expression on the plasma membrane. GCaMP6s was used to functionally analyze Rpsa knockdown via ex vivo calcium imaging. Rpsa-deficient neurons showed less fluctuation in fluorescence intensity, suggesting defective subthreshold calcium signaling. The Serpinf1 gene coding for PEDF is localized at chromosome 17p13.3, which is deleted in patients with the neurodevelopmental disorder Miller-Dieker syndrome. Our study identifies a role for Rpsa in early cortical development and for PEDF-Rpsa-Itga6 signaling in neuromorphogenesis, thus implicating these molecules in the etiology of neurodevelopmental disorders like Miller-Dieker syndrome and identifying them as potential therapeutics.