核受体NR4A1基因前脑特异性敲除小鼠的构建及行为学检测
Construction and behavioral detection of forebrain-specific nuclear receptor NR4A1 gene knockout mice
摘要
Abstract
Objective To construct a forebrain excitatory neuron-specific NR4A1 knockout mouse model based on the LoxP-Cre recombinase system and to investigate the effects of this knockout on cognitive function and anxiety-like behaviors in mice.Methods NR4A1flox/+mice were self-crossed and screened to obtain NR4A1flox/flox mice,which were then crossed with Camk2a-Cre mice.NR4A1flox/+-Camk2a-Cre F1 mice were obtained and further self-crossed.The result ing offspring were genotyped using tail genomic DNA to screen out NR4A1flox/flox-Camk2a-Cre mice,which were defined as NR4A1 conditional knockout mice,specifically in forebrain excitatory neurons.Cognitive ability was assessed using behavioral tests,including novel object recognition and Y-maze tests,and anxiety-like behaviors were evaluated using the elevated plus maze,open field,and light-dark box tests.Expression levels of NR4A1 in the hippocampus,as a key forebrain region,were detected by Western Blot and immunofluorescence.Results(1)Genotyping confirmed that NR4A1flox/flox-Camk2a-Cre forebrain excitatory neuron-specific NR4A1 knockout mice were successfully obtained.(2)NR4A1 protein levels were significantly reduced in the hippocampus in knockout mice compared with the findings in control mice,as shown by Western Blot.Also,the intensity of NR4A1 immunofluorescent signals in the hippocampus was also lower in knockout group compared with that in control group mice.(3)In cognitive behavioral tests,there was no significant difference in novel object recognition index,number of entries,distance traveled,or residence time in the novel arm of the Y-maze between knockout group and control group mice.(4)In anxiety-like behavior tests,knockout group mice spent more time in the central area of the open field than control group mice,but there was no significant difference in the distance traveled or number of entries into the central area.In the light-dark box test,knockout group mice spent longer in the light compartment than control group mice,but there was no significant difference in the distance traveled or number of entries into the light compartment between the two groups.There was no significant difference between knockout group and control group mice in the elevated plus maze test.Conclusions This study successfully constructed a forebrain excitatory neuron-specific NR4A1 knockout mouse model.Knockout mice showed no significant changes in cognitive level but a significant reduction in anxiety levels.This model provides an important experimental tool for in-depth investigations of the role of the NR4A1 gene in the physiological functions and pathological mechanisms of forebrain regions.关键词
核受体NR4A1/海马/条件性敲除小鼠模型/认知水平/焦虑样行为Key words
nuclear receptor NR4A1/hippocampus/conditional knockout mouse model/cognitive level/anxiety-like behavior分类
生物科学引用本文复制引用
王紫君,白雁中,安文好,秦豪泽,葛芯雨,刘丽娜,王传升,张瑞岭,张艳杰..核受体NR4A1基因前脑特异性敲除小鼠的构建及行为学检测[J].中国实验动物学报,2025,33(12):1749-1758,10.基金项目
河南省科技研发计划联合基金(产业化)重大项目(235101610004),河南省医学科技攻关(联合共建)项目(LHGJ20230532),河南省自然科学基金面上项目(252300421398),河南省自然科学基金青年项目(242300421539),河南省重点研发与推广专项科技攻关(242102310286),河南省生物精神病学重点实验室开放课题(ZDSYS2022005),河南省大学生创新创业训练计划项目(202410472022). Funded by Henan Province Science and Technology Research and Development Plan Joint Fund(Industry)Major Project(235101610004),Henan Provincial Medical Science and Technology Tackling Key Problems(Jointly Constructed)Project(LHGJ20230532),General Grant Program of Henan Provincial Natural Science Foundation(252300421398),Henan Provincial Natural Science Foundation Youth Project(242300421539),Henan Provincial Key R&D and Promotion Special Program for Technological Breakthroughs(242102310286),the Open Project of Henan Key Laboratory of Biological Psychiatry(ZDSYS2022005),Henan Province College Student Innovation and Entrepreneurship Training Program(202410472022). （产业化）
