Abstract：
OBJECTIVE: Zinc finger protein 804A (ZNF804A) was the first genome-wide associated susceptibility gene for schizophrenia (SCZ) and played an essential role in the pathophysiology of SCZ by influencing neurodevelopment regulation, neurite outgrowth, synaptic plasticity, and RNA translational control; however, the exact molecular mechanism remains unclear.
METHODS: A nervous-system-specific Zfp804a (ZNF804A murine gene) conditional knockout (cKO) mouse model was generated using clustered regularly interspaced short palindromic repeat/Cas9 technology and the Cre/loxP method.
RESULTS: Multiple and complex SCZ-like behaviors, such as anxiety, depression, and impaired cognition, were observed in Zfp804a cKO mice. Molecular biological methods and targeted metabolomics assay validated that Zfp804a cKO mice displayed altered SATB2 (a cortical superficial neuron marker) expression in the cortex; aberrant NeuN, cleaved caspase 3, and DLG4 (markers of mature neurons, apoptosis, and postsynapse, respectively) expressions in the hippocampus and a loss of glutamate (Glu)/γ-aminobutyric acid (GABA) homeostasis with abnormal GAD67 (Gad1) expression in the hippocampus. Clozapine partly ameliorated some SCZ-like behaviors, reversed the disequilibrium of the Glu/GABA ratio, and recovered the expression of GAD67 in cKO mice.
CONCLUSIONS: Zfp804a cKO mice reproducing SCZ-like pathological and behavioral phenotypes were successfully developed. A novel mechanism was determined in which Zfp804a caused Glu/GABA imbalance and reduced GAD67 expression, which was partly recovered by clozapine treatment. These findings underscore the role of altered gene expression in understanding the pathogenesis of SCZ and provide a reliable SCZ model for future therapeutic interventions and biomarker discovery.
METHODS: A nervous-system-specific Zfp804a (ZNF804A murine gene) conditional knockout (cKO) mouse model was generated using clustered regularly interspaced short palindromic repeat/Cas9 technology and the Cre/loxP method.
RESULTS: Multiple and complex SCZ-like behaviors, such as anxiety, depression, and impaired cognition, were observed in Zfp804a cKO mice. Molecular biological methods and targeted metabolomics assay validated that Zfp804a cKO mice displayed altered SATB2 (a cortical superficial neuron marker) expression in the cortex; aberrant NeuN, cleaved caspase 3, and DLG4 (markers of mature neurons, apoptosis, and postsynapse, respectively) expressions in the hippocampus and a loss of glutamate (Glu)/γ-aminobutyric acid (GABA) homeostasis with abnormal GAD67 (Gad1) expression in the hippocampus. Clozapine partly ameliorated some SCZ-like behaviors, reversed the disequilibrium of the Glu/GABA ratio, and recovered the expression of GAD67 in cKO mice.
CONCLUSIONS: Zfp804a cKO mice reproducing SCZ-like pathological and behavioral phenotypes were successfully developed. A novel mechanism was determined in which Zfp804a caused Glu/GABA imbalance and reduced GAD67 expression, which was partly recovered by clozapine treatment. These findings underscore the role of altered gene expression in understanding the pathogenesis of SCZ and provide a reliable SCZ model for future therapeutic interventions and biomarker discovery.
摘要：
目的：锌指蛋白804A（ZNF804A）是第一个全基因组相关的精神分裂症易感基因（SCZ），通过影响神经发育调节在SCZ的病理生理学中起重要作用。神经突生长，突触可塑性，和RNA翻译控制；然而，确切的分子机制尚不清楚。
方法：使用成簇的规则间隔短回文重复/Cas9技术和Cre/loxP方法产生了神经系统特异性Zfp804a（ZNF804A鼠基因）条件敲除（cKO）小鼠模型。
结果：多种复杂的类似SCZ的行为，比如焦虑，抑郁症,和认知受损，在Zfp804acKO小鼠中观察到。分子生物学方法和靶向代谢组学测定验证了Zfp804acKO小鼠在皮质中显示出SATB2（皮质浅表神经元标记）表达的改变；异常的NeuN，裂解的胱天蛋白酶3和DLG4(成熟神经元的标记，凋亡，和突触后，分别）海马中的表达和海马中GAD67（Gad1）表达异常的谷氨酸（Glu）/γ-氨基丁酸（GABA）稳态丧失。氯氮平部分改善了一些SCZ样行为，逆转了Glu/GABA比例的不平衡，并恢复了cKO小鼠中GAD67的表达。
结论：Zfp804acKO小鼠成功复制了SCZ样病理和行为表型。确定了一种新机制，其中Zfp804a引起Glu/GABA失衡并降低GAD67表达，部分通过氯氮平治疗恢复。这些发现强调了基因表达改变在理解SCZ发病机制中的作用，并为未来的治疗干预和生物标志物发现提供了可靠的SCZ模型。
方法：使用成簇的规则间隔短回文重复/Cas9技术和Cre/loxP方法产生了神经系统特异性Zfp804a（ZNF804A鼠基因）条件敲除（cKO）小鼠模型。
结果：多种复杂的类似SCZ的行为，比如焦虑，抑郁症,和认知受损，在Zfp804acKO小鼠中观察到。分子生物学方法和靶向代谢组学测定验证了Zfp804acKO小鼠在皮质中显示出SATB2（皮质浅表神经元标记）表达的改变；异常的NeuN，裂解的胱天蛋白酶3和DLG4(成熟神经元的标记，凋亡，和突触后，分别）海马中的表达和海马中GAD67（Gad1）表达异常的谷氨酸（Glu）/γ-氨基丁酸（GABA）稳态丧失。氯氮平部分改善了一些SCZ样行为，逆转了Glu/GABA比例的不平衡，并恢复了cKO小鼠中GAD67的表达。
结论：Zfp804acKO小鼠成功复制了SCZ样病理和行为表型。确定了一种新机制，其中Zfp804a引起Glu/GABA失衡并降低GAD67表达，部分通过氯氮平治疗恢复。这些发现强调了基因表达改变在理解SCZ发病机制中的作用，并为未来的治疗干预和生物标志物发现提供了可靠的SCZ模型。
