背景:已知神经发育障碍相关基因,Satb2在确定上层神经元规范中起着重要作用。然而,尚不清楚该基因在发育过程中如何调节其他新皮质区域。也缺乏对其在新皮层发育中的空间调节途径的全面描述。
结果:在这项工作中,我们利用空间转录组学和免疫染色,通过比较Satb2+/+和Satb2-/-小鼠在胚胎阶段,系统地研究了Satb2的区域特异性基因调控。包括心室区(VZ)或心室下区(SVZ),中间区(IZ)和皮质板(CP)。染色结果显示,这三个区域在Satb2-/-小鼠中变得中等或显著更薄。在细胞层面,细胞数量在VZ/SVZ中增加,而CP中的细胞数量减少。空间转录组学数据表明,许多重要的基因和相关途径在Satb2-/-小鼠中以区域特异性方式失调。在VZ/SVZ中,参与神经前体细胞增殖的关键基因,包括中间祖细胞标记Tbr2和乳酸产生相关基因Ldha,在Satb2-/-小鼠中上调。在IZ,调节神经元分化和迁移的关键基因,如Rnd2,在Satb2-/-小鼠中表现出异位表达。在CP中,谱系特异性基因,Tbr1和Bcl11b,表达异常。神经肽相关基因Npy在Satb2-/-小鼠中下调。最后,我们通过免疫荧光或qPCR验证了关键调节因子的异常表达。
结论:总之,我们的工作提供了有关Satb2在新皮质发育中调控的区域特异性基因和途径的见解。
BACKGROUND: It is known that the neurodevelopmental disorder associated gene, Satb2, plays important roles in determining the upper layer neuron specification. However, it is not well known how this gene regulates other neocortical regions during the development. It is also lack of comprehensive delineation of its spatially regulatory pathways in neocortical development.
RESULTS: In this work, we utilized spatial transcriptomics and immuno-staining to systematically investigate the region-specific gene regulation of Satb2 by comparing the Satb2+/+ and Satb2-/- mice at embryonic stages, including the ventricle zone (VZ) or subventricle zone (SVZ), intermediate zone (IZ) and cortical plate (CP) respectively. The staining result reveals that these three regions become moderately or significantly thinner in the Satb2-/- mice. In the cellular level, the cell number increases in the VZ/SVZ, whereas the cell number decreases in the CP. The spatial transcriptomics data show that many important genes and relevant pathways are dysregulated in Satb2-/- mice in a region-specific manner. In the VZ/SVZ, the key genes involved in neural precursor cell proliferation, including the intermediate progenitor marker Tbr2 and the lactate production related gene Ldha, are up-regulated in Satb2-/- mice. In the IZ, the key genes in regulating neuronal differentiation and migration, such as Rnd2, exhibit ectopic expressions in the Satb2-/- mice. In the CP, the lineage-specific genes, Tbr1 and Bcl11b, are abnormally expressed. The neuropeptide related gene Npy is down-regulated in Satb2-/- mice. Finally, we validated the abnormal expressions of key regulators by using immunofluorescence or qPCR.
CONCLUSIONS: In summary, our work provides insights on the region-specific genes and pathways which are regulated by Satb2 in neocortical development.