Jiang Wei, Zhang Jian, Wang Molin, Zou Yongxin, Liu Qiao, Song Yu, Sun Gongping, Gong Yaoqin, Zhang Fan, Jiang Baichun
The Key Laboratory of Experimental Teratology of the Ministry of Education and Department of Genetics, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
The Key Laboratory of Experimental Teratology of the Ministry of Education, Department of Histology and Embryology, School of Basic Medical Sciences, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, China.
Acta Neuropathol Commun. 2024 Dec 4;12(1):188. doi: 10.1186/s40478-024-01903-y.
Cullin 4B (CUL4B) is the scaffold protein in the CUL4B-RING E3 ubiquitin ligase (CRL4B) complex. Loss-of-function mutations in the human CUL4B gene lead to syndromic X-linked intellectual disability (XLID). Till now, the mechanism of intellectual disability caused by CUL4B mutation still needs to be elucidated. In this study, we used single-nucleus RNA sequencing (snRNA-seq) to investigate the impact of CUL4B deficiency on the transcriptional programs of diverse cell types. The results revealed that depletion of CUL4B resulted in impaired intercellular communication and elicited cell type-specific transcriptional changes relevant to synapse dysfunction. Golgi-Cox staining of brain slices and immunostaining of in vitro cultured neurons revealed remarkable synapse loss in CUL4B-deficient mice. Ultrastructural analysis via transmission electron microscopy (TEM) showed that the width of the synaptic cleft was significantly greater in CUL4B-deficient mice. Electrophysiological experiments found a decrease in the amplitude of AMPA receptor-mediated EPSCs in the hippocampal CA1 pyramidal neurons of CUL4B-deficient mice. These results indicate that depletion of CUL4B in mice results in morphological and functional abnormalities in synapses. Furthermore, behavioral tests revealed that depletion of CUL4B in the mouse nervous system results in impaired spatial learning and memory. Taken together, the findings of this study reveal the pathogenesis of neurological disorders associated with CUL4B mutations and promote the identification of therapeutic targets that can halt synaptic abnormalities and preserve memory in individuals.
Cullin 4B(CUL4B)是CUL4B-RING E3泛素连接酶(CRL4B)复合物中的支架蛋白。人类CUL4B基因的功能丧失突变会导致综合征性X连锁智力障碍(XLID)。到目前为止,CUL4B突变导致智力障碍的机制仍有待阐明。在本研究中,我们使用单核RNA测序(snRNA-seq)来研究CUL4B缺陷对多种细胞类型转录程序的影响。结果显示,CUL4B的缺失导致细胞间通讯受损,并引发与突触功能障碍相关的细胞类型特异性转录变化。脑片的高尔基-考克斯染色和体外培养神经元的免疫染色显示,CUL4B缺陷小鼠存在明显的突触丢失。通过透射电子显微镜(TEM)进行的超微结构分析表明,CUL4B缺陷小鼠的突触间隙宽度明显更大。电生理实验发现,CUL4B缺陷小鼠海马CA1锥体神经元中AMPA受体介导的兴奋性突触后电流(EPSC)幅度降低。这些结果表明,小鼠中CUL4B的缺失导致突触出现形态和功能异常。此外,行为测试显示,小鼠神经系统中CUL4B的缺失导致空间学习和记忆受损。综上所述,本研究结果揭示了与CUL4B突变相关的神经疾病的发病机制,并促进了对能够阻止突触异常并保护个体记忆的治疗靶点的识别。