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Distinct neuronal excitability alterations of medial prefrontal cortex in early-life neglect model of rats

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Animal Models and Experimental Medicine 2022年第3期5卷 274-280页

作者： Yu Zhang Xiuping Sun Changsong Dou Xianglei Li Ling Zhang Chuan Qin NHC Key Laboratory of Human Disease Comparative Medicine Institute of Laboratory Animal SciencesChinese Academy of Medical Sciences(CAMS)Comparative Medicine CenterPeking Union Medical College(PUMC)BeijingChina National Human Diseases Animal Model Resource Center BeijingChina Beijing Engineering Research Center for Experimental Animal Models of Human Critical Diseases BeijingChina International Center for Technology and Innovation of animal model BeijingChina Changping National laboratory(CPNL) BeijingChina

Object:Early-life neglect has irreversible emotional effects on the central nervous system.In this work,we aimed to elucidate distinct functional neural changes in me-dial prefrontal cortex(mPFC)of model rats.Methods:Maternal separation with early weaning was used as a rat model of early-life neglect.The excitation of glutamatergic and GABAergic neurons in rat mPFC was recorded and analyzed by whole-cell patch clamp.Results:Glutamatergic and GABAergic neurons of mPFC were distinguished by typi-cal electrophysiological properties.The excitation of mPFC glutamatergic neurons was significantly increased in male groups,while the excitation of mPFC GABAergic neurons was significant in both female and male groups,but mainly in terms of rest membrane potential and amplitude,respectively.Conclusions:Glutamatergic and GABAergic neurons in medial prefrontal cortex showed different excitability changes in a rat model of early-life neglect,which can contribute to distinct mechanisms for emotional and cognitive manifestations.

关键词： early-life neglect model GABAergic glutamatergic maternal separation with early weaning medial prefrontal cortex neuronal excitability

A genome-wide identification of the BLH gene family reveals BLH1 involved in cotton fiber development

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Journal of Cotton Research 2020年第3期3卷 199-209页

作者： LIU Cuixia LI Zhifang dou Lingling YUAN Yi ZOU changsong SHANG Haihong CUI Langjun XIAO Guanghui Key Laboratory of the Ministry of Education for Medicinal Plant Resources and Natural Pharmaceutical Chemistry National Engineering Laboratory for Resouce Development of Endangered Crude Drugs in the Northwest of ChinaCollege of Life SciencesShaanxi Normal UniversityXi’an 710119ShaanxiChina College of Biology Pharmacy and Food Engineering Shangluo UniversityShangluo 726000ShaanxiChina State Key Laboratory of Cotton Biology State Key Laboratory of Crop Stress Adaptation and ImprovementSchool of Life SciencesHenan UniversityKaifeng 475001HenanChina School of Chemistry and Chemical Engineering Xianyang Normal UniversityXianyang 712000ShaanxiChina Zhengzhou Research Base State Key Laboratory of Cotton BiologyZhengzhou UniversityZhengzhou 450001China

Background:Cotton is the world’s largest and most important source of renewable natural fiber.BEL1-like homeodomain(BLH)genes are ubiquitous in plants and have been reported to contribute to plant development.However,there is no comprehensive characterization of this gene family in cotton.In this study,32,16,and 18 BLH genes were identified from the G.hirsutum,G.arboreum,and G.raimondii genome,respectively.In addition,we also studied the phylogenetic relationships,chromosomal location,gene structure,and gene expression patterns of the BLH genes.Results:The results indicated that these BLH proteins were divided into seven distinct groups by phylogenetic analysis.Among them,25 members were assigned to 15 chromosomes.Furthermore,gene structure,chromosomal location,conserved motifs,and expression level of BLH genes were investigated in G.hirsutum.Expression profiles analysis showed that four genes(GhBLH1_3,GhBLH1_4,GhBLH1_5,and GhBLH1_6)from BLH1 subfamily were highly expressed during the fiber cell elongation period.The expression levels of these genes were significantly induced by gibberellic acid and brassinosteroid,but not auxin.Exogenous application of gibberellic acid significantly enhanced GhBLH1_3,GhBLH1_4,and GhBLH1_5 transcripts.Expression levels of GhBLH1_3 and GhBLH1_4 genes were significantly increased under brassinosteroid treatment.Conclusions:The BLH gene family plays a very important role in many biological processes during plant growth and development.This study deepens our understanding of the role of the GhBLH1 gene involved in fiber development and will help us in breeding better cotton varieties in the future.

关键词： Cotton G.hirsutum BLH gene family Fiber development

黑质LAMP2A过表达改善帕金森病恒河猴模型的运动功能

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中国比较医学杂志 2022年第4期32卷 7-13页

作者：朱旭超窦长松张钰秦川张玲中国医学科学院医学实验动物研究所北京协和医学院比较医学中心国家卫生健康委员会人类疾病比较医学重点实验室国家中医药管理局人类动物模型三级实验室北京100021

目的在帕金森病(Parkinson’s disease,PD)恒河猴模型上观察黑质过表达溶酶体相关膜蛋白2A(lysosome associated membrane protein 2A,LAMP2A)上调分子伴侣介导的自噬(chaperone-mediated autophagy,CMA)对实验猴运动行为的影响。方法应用立体定位注射仪向7只恒河猴的右脑纹状体区注射PD患者死亡后脑组织(自愿捐献)的路易小体(Lewy bodies,LBs)提取物,同时在左脑纹状体区注射非PD病人死亡患者的脑组织(自愿捐献)提取物来构建PD恒河猴模型。模型动物随机分为两组,LAMP2A过表达组(4只)在其黑质区双侧注射过表达LAMP2A的腺相关病毒载体;对照组(3只)在黑质区双侧注射对照病毒载体。通过阶梯实验、平板取食实验评价PD恒河猴模型运动能力和LAMP2A对该PD模型运动功能的影响。利用ELISA法对实验猴脑脊液、血清的总α-突触核蛋白(α-synuclein,α-syn)及磷酸化α-syn水平进行测定。结果阶梯实验和平板取食实验中对照组恒河猴的左侧上肢的精细动作水平相较于其右侧出现下降;LAMP2A过表达组恒河猴的左右两侧上肢的精细运动水平没有出现明显差异;ELISA测定对照组术后12月脑脊液、血清的总α-syn水平及磷酸化α-syn水平显著增高;LAMP2A过表达组脑脊液磷酸化α-syn水平和血清磷酸化α-syn水平在8月之后均低于同期的对照组。结论过表达LAMP2A可能通过增加α-syn的CMA清除改善PD恒河猴模型精细运动能力。

关键词： LAMP2A 帕金森病行为学恒河猴

Genome-wide identification and expression analysis of the GhIQD gene family in upland cotton(Gossypium hirsutum L.)

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Journal of Cotton Research 2021年第1期4卷 24-37页

作者： dou Lingling LV Limin KANG Yangyang TIAN Ruijie HUANG Deqing LI Jiayin LI Siyi LIU Fengping CAO Lingyan JIN Yuhua LIU Yang LI Huaizhu WANG Wenbo PANG Chaoyou SHANG Haihong ZOU changsong SONG Guoli XIAO Guanghui School of Chemistry and Chemical Engineering Xianyang Normal UniversityXianyang 712000ShaanxiChina State Key Laboratory of Cotton Biology Institute of Cotton Research of Chinese Academy of Agricultural SciencesAnyang 455000China Zhengzhou University research base for State Key Laboratory of Cotton Biology in China ZhengzhouChina Key Laboratory of Plant Stress Biology State Key Laboratory of Cotton BiologySchool of Life SciencesHenan UniversityKaifengChina College of Life Sciences Shaanxi Normal UniversityXi’an 710119China

Background:Calmodulin(CaM)is one of the most important Ca^(2+)signaling receptors because it regulates diverse physiological and biochemical reactions in plants.CaM functions by interacting with CaM-binding proteins(CaMBPs)to modulate Ca^(2+)signaling.IQ domain(IQD)proteins are plant-specific CaMBPs that bind to CaM by their specific CaM binding sites.Results:In this study,we identified 102 GhIQD genes in the Gossypium hirsutum L.genome.The GhIQD gene family was classified into four clusters(Ⅰ,Ⅱ,Ⅲ,andⅣ),and we then mapped the GhIQD genes to the G.hirsutum L.chromosomes.Moreover,we found that 100 of the 102 GhIQD genes resulted from segmental duplication events,indicating that segmental duplication is the main force driving GhIQD gene expansion.Gene expression pattern analysis showed that a total of 89 GhIQD genes expressed in the elongation stage and second cell wall biosynthesis stage of the fiber cells,suggesting that GhIQD genes may contribute to fiber cell development in cotton.In addition,we found that 20 selected GhIQD genes were highly expressed in various tissues.Exogenous application of MeJA significantly enhanced the expression levels of GhIQD genes.Conclusions:Our study shows that GhIQD genes are involved in fiber cell development in cotton and are also widely induced by MeJA.Thw results provide bases to systematically characterize the evolution and biological functions of GhIQD genes,as well as clues to breed better cotton varieties in the future.

关键词： Gossypium hirsutum L. GhIQD genes Segmental duplication Expression analysis

Patterns of presence-absence variants in Upland cotton

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Science China(Life Sciences) 2020年第10期63卷 1600-1603页

作者： Junfang Liu Xianliang Zhang Lingling dou Wei Li Xiaojian Zhou Yangai Liu Xiaoyu Pei Zhongying Ren Wensheng Zhang Huaizhu Li Wenbo Wang changsong Zou Kunlun He Fei Zhang Wenyu Ma Haihong Shang Hongbin Li Daigang Yang Guanghui Xiao Xiongfeng Ma State Key Laboratory of Cotton Biology Institute of Cotton ResearchChinese Academy of Agricultural SciencesAnyang 455000China School of Chemistry and Chemical Engineering Xianyang Normal UniversityXianyang 712000China Key Laboratory of Plant Stress Biology State Key Laboratory of Cotton BiologySchool of Life SciencesHenan UniversityKaifeng 475004China Key Laboratory of the Ministry of Education for Medicinal Plant Resources and Natural Pharmaceutical Chemistry National Engineering Laboratory for Resource Development of Endangered Crude Drugs in the Northwest of ChinaCollege of Life SciencesShaanxi Normal UniversityXi’an 710119China Zhengzhou Research Base State Key Laboratory of Cotton BiologyZhengzhou UniversityZhengzhou 450001China College of Life Sciences Shihezi UniversityShihezi 832003China

Dear Editor,Sources of genetic variations in genomes include small-scale sources(such as single-nucleotide polymorphisms(SNPs),insertions/deletions(InDels),and simple sequence repeats and larger-scale structural variations(mainly presence-absence variants(PAVs))and copy number variants).PAVs are sequences that are either inserted or missing in genomes in comparison with a reference sequence or genome.PAVs can have a much longer sequence than SNPs and InDels,as illustrated in the human genome(Conrad et al.,2010).PAVs are important genomic structural variations that can directly affect genomic structure and key functional genes in the genome(Kumar et al.,2007).Moreover,the use of PAVs for studying quantitative traits has been valuable(Lam et al.,2010).

关键词： traits sequence insertion

锰中毒和PD模型小鼠神经行为改变及损伤部位的比较研究

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中华劳动卫生职业病杂志 2018年第2期36卷 84-90页

作者：窦长松智翠娜刘文丽付娟玲姚碧云北京大学公共卫生学院毒理系、食品安全毒理学研究与评价北京市重点实验室 100191

目的探讨锰中毒和PD模型小鼠神经行为学及组织病理学的异同，为锰中毒的诊断和防治提供依据。方法成年C57BL/6雄性小鼠共80只，随机分为5组，每组16只，MnCl2低、中、高剂量组分别腹腔注射5、10、20 mg/kg MnCl2，PD模型组腹腔注射30 mg... 详细信息

目的探讨锰中毒和PD模型小鼠神经行为学及组织病理学的异同，为锰中毒的诊断和防治提供依据。方法成年C57BL/6雄性小鼠共80只，随机分为5组，每组16只，MnCl2低、中、高剂量组分别腹腔注射5、10、20 mg/kg MnCl2，PD模型组腹腔注射30 mg/kg MPTP，对照组注射等量生理盐水，5次/周，连续4周，每天观察动物一般情况。注射结束1周后，通过爬杆实验、旷场实验、水迷宫实验检测小鼠平衡协调能力、焦虑抑郁状态以及学习记忆等神经行为学改变；处死动物，显微镜下观察纹状体及黑质H-E染色病理切片，检查神经细胞病理变化。结果与对照组比较，MnCl2高剂量组小鼠体重明显降低，差异有统计学意义（P〈0.01）。与对照组比较，MnCl2和MPTP组小鼠爬杆实验总时间和转身时间明显增加差异均有统计学意义（P〈0.05），MnCl2中、高剂量组和MPTP组小鼠旷场实验穿过格子数、站立次数以及中央场活动时间明显减少（P〈0.05），MnCl2高剂量组及MPTP组小鼠水迷宫实验逃避潜伏时间增加且目标象限活动时间减少，差异均有统计学意义（P〈0.05）。与MPTP组比较，MnCl2高剂量组逃避潜伏时间明显增加，差异有统计学意义（P〈0.05）；病理结果显示，与对照组比较，MnCl2及MPTP组嗜酸性水平增高，其中纹状体嗜酸性细胞数MnCl2高剂量组高于MPTP组，差异有统计学意义（P〈0.01），黑质嗜酸性细胞MnCl2组少于MPTP组，差异有统计学意义（P〈0.01）。结论锰和MPTP均可引起多巴胺能神经系统损伤，锰中毒导致的神经行为学改变及损伤部位与MPTP诱导的PD模型不完全一致。

关键词：锰 1-甲基-4-苯基-1 2 3 6-四氢吡啶神经行为学纹状体黑质 C57BL/6小鼠

溶酶体在锰致SK-N-SH细胞毒性中的作用

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中华劳动卫生职业病杂志 2019年第5期37卷 332-336页

作者：智翠娜来丽叶窦长松王雪珩赵鹏傅娟玲姚碧云北京大学公共卫生学院毒理系食品安全毒理学研究与评价北京市重点实验室100191 北京大学基础医学院 100191

目的探讨溶酶体在锰致人神经母细胞瘤(SK-N-SH)细胞毒性中的作用。方法以浓度分别为0、0.0625、0.125、0.25、0.5、1.0、2.0和4.0mmol/LMnCl2处理SK-N-SH细胞24h,通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞存活率。... 详细信息

目的探讨溶酶体在锰致人神经母细胞瘤(SK-N-SH)细胞毒性中的作用。方法以浓度分别为0、0.0625、0.125、0.25、0.5、1.0、2.0和4.0mmol/LMnCl2处理SK-N-SH细胞24h,通过3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法检测细胞存活率。以0、0.125、0.25、0.5和1.0mmol/LMnCl2处理细胞24h,激光共聚焦显微镜观察红色荧光探针标记的溶酶体,蛋白免疫印迹(Westernblot)法检测溶酶体膜相关蛋白1(LAMP1)、组织蛋白酶D(CTSD)表达水平,CTSD活力荧光测定试剂盒检测CTSD活力。结果与对照组比较,0.5~4.0mmol/LMnCl2处理组SK-N-SH细胞存活率明显降低,0.5~1.0mmol/LMnCl2处理组SK-N-SH细胞相对荧光强度明显增加,差异均有统计学意义(P0.05)。结论MnCl2可引起SK-N-SH细胞毒性,溶酶体在锰低剂量时发挥正常功能,但在锰高剂量时受损。溶酶体作为自噬下游发挥降解功能的细胞器,可能参与了锰的神经毒性机制。

关键词：锰神经母细胞瘤细胞溶酶体溶酶体膜相关蛋白1 组织蛋白酶D

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飞机超低量喷洒噻虫啉防治松褐天牛应用报告

安徽林业科技 2019年第1期45卷 13-15,23页

作者：解全荣吴庆杰郭晋保刘伟炜窦长松山东昆丰农林科技股份有限公司山东潍坊261000

松材线虫病是松树的毁灭性病害,防治松褐天牛,切断松材线虫传播途径是防治的主要措施之一。使用飞机进行超低量喷洒2%噻虫啉微囊悬浮剂,分两次对李沧区松材线虫病进行防治,两次防治面积共5333hm^2。结果表明,该药剂对天牛成虫的防治效... 详细信息

松材线虫病是松树的毁灭性病害,防治松褐天牛,切断松材线虫传播途径是防治的主要措施之一。使用飞机进行超低量喷洒2%噻虫啉微囊悬浮剂,分两次对李沧区松材线虫病进行防治,两次防治面积共5333hm^2。结果表明,该药剂对天牛成虫的防治效果平均为90%以上,对松材线虫病的防治效果达到了66.67%。

关键词：松材线虫病松褐天牛超低量飞防噻虫啉

MnCl2和MPP^＋诱导SK-N-SH细胞氧化应激及自噬的比较

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中华劳动卫生职业病杂志 2017年第2期35卷 96-100页

作者：刘文丽窦长松王裕赵鹏傅娟玲姚碧云周宗灿北京大学公共卫生学院食品安全毒理学研究与评价北京市重点实验室100191

目的研究锰和1-甲基-4-苯基吡啶（MPP^＋）引起人神经母细胞瘤（SK-N-SH）细胞氧化应激和自噬的异同,进一步探讨锰神经毒性机制.方法以0.0625、0.125、0.25、0.5、1.0和2.0 mmol/L MnCl2和MPP^＋分别处理SK-N-SH细胞24 h,采用3-（4,5... 详细信息

目的研究锰和1-甲基-4-苯基吡啶（MPP^＋）引起人神经母细胞瘤（SK-N-SH）细胞氧化应激和自噬的异同,进一步探讨锰神经毒性机制.方法以0.0625、0.125、0.25、0.5、1.0和2.0 mmol/L MnCl2和MPP^＋分别处理SK-N-SH细胞24 h,采用3-（4,5-二甲基噻唑-2）-2,5-二苯基四氮唑溴盐（MTT）试验检测细胞存活率;0.125、0.25、0.5 mmol/L MnCl2和MPP^＋分别处理细胞24 h,采用流式细胞仪检测细胞内活性氧（ROS）含量,激光共聚焦显微镜观察细胞自噬体,蛋白免疫印迹（Western blotting）法检测自噬相关蛋白P62和LC3-Ⅱ/LC3-Ⅰ表达.结果与对照组比较,0.0625-2.0 mmol/L MnCl2和0.125-2.0 mmol/L MPP^＋处理组SK-N-SH细胞存活率均明显降低,且0.25-2.0 mmol/L MnCl2处理组SK-N-SH细胞存活率明显低于相同剂量MPP^＋处理组（均P〈0.05）;与对照组比较,0.125-0.25 mmol/L MnCl2和0.125-0.5 mmol/L MPP^＋处理组SK-N-SH细胞内ROS含量均明显增加,0.25-0.5 mmol/L MPP^＋处理组细胞内ROS含量明显高于相同剂量MnCl2处理组（均P〈0.05）;与对照组比较,0.25-0.5 mmol/L MnCl2和MPP^＋处理组细胞自噬相关蛋白LC3-Ⅱ/LC3-Ⅰ值均明显增加,P62表达均明显下降,且0.125-0.5 mmol/L MPP^＋处理组LC3-Ⅱ/LC3-Ⅰ值明显高于相同剂量MnCl2处理组,0.125和0.25 mmol/L MPP^＋处理组P62表达水平明显低于相同剂量MnCl2处理组（均P〈0.05）.结论 MnCl2和MPP^＋均可引起SK-N-SH细胞氧化应激,诱导自噬,且MPP^＋对SK-N-SH细胞自噬的诱导作用明显高于MnCl2.

关键词：锰 1-甲基-4-苯基吡啶自噬氧化应激 SK-N-SH细胞 MPP+