,1, 张明1, 刘瑞莹1, 万贵钧1, 潘卫东2, 陈法军,1Effects of the Interference of Key Magnetic Response Genes on the Longevity of Brown Planthopper (Nilaparvata lugens) Under Near-Zero Magnetic Field
HE JingLan,1, ZHANG Ming1, LIU RuiYing1, WAN GuiJun1, PAN WeiDong2, CHEN FaJun,1通讯作者:
第一联系人:
收稿日期:2018-08-2接受日期:2018-09-11网络出版日期:2019-01-01
| 基金资助: |
Received:2018-08-2Accepted:2018-09-11Online:2019-01-01
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贺静澜, 张明, 刘瑞莹, 万贵钧, 潘卫东, 陈法军. 近零磁场下干扰磁响应关键基因对褐飞虱寿命的影响[J]. 中国农业科学, 2019, 52(1): 45-55 doi:10.3864/j.issn.0578-1752.2019.01.005
HE JingLan, ZHANG Ming, LIU RuiYing, WAN GuiJun, PAN WeiDong, CHEN FaJun.
0 引言
【研究意义】磁场能以机械力的方式作用于一定范围内的磁体或运动电荷[1]。根据磁场的强度（用磁感应强度B表示,单位为特斯拉,T）,可人为地将磁场划分为弱磁场（<1 mT）、中等强度磁场（1 mT—1 T）、强磁场（1—5 T）和超强磁场（>5 T）[2]。地球磁场（geomagnetic field,GMF）的平均场强为50 μT左右,属于弱磁场范围,而强度比地磁场更小的弱磁环境又可进一步被定义为亚磁场（亚磁空间）,也可称近零磁场（near-zero magnetic field,NZMF）[3]。生物磁响应（magnetic response）指生物感知磁场强度及方向的信息,并直接或间接地通过行为、生理、代谢等作出从宏观表型到微观分子水平响应的现象[4]。目前对于弱磁场,尤其是地磁场,人们更侧重于研究其对生物行为的影响,如以家鸽为代表的迁徙性动物极有可能利用自地磁信息进行定向和导航[5];至于强磁场和超强磁场,人们则更关注这类磁场对人体造成的潜在威胁,如核磁共振成像时的短时强磁空间等[6];因此磁场生物学效应的研究一度集中于中等强度磁场[7];而随着近千年来地球磁场的不断衰弱,针对强度低于地磁场强度——即近零磁场的生物磁响应研究也逐渐增多。实验证实,如果长期处于该磁场环境下,动物的中枢神经系统将产生障碍[8,9,10,11,12],早期发育过程受到明显影响[13,14],而短期暴露近零磁场环境也会影响人体的健康[15,16,17]。可见进行有关近零磁场下生物的磁生理响应研究,进而推测地球磁场的变化对生物的不利效应具有重要意义。【前人研究进展】大量研究有力地证实了生物可对磁场变化作出响应,而相较于此,对于生物磁响应机制的探索却一直未有定论。事实上,生物对地磁场感知的关键在于“磁受体”（magnetoreceptor）的确定,对此****们提出了多种假说,而其中有两种得到了广泛认可,即磁颗粒介导的磁受体假说与依赖光的自由基对假说。前者认为,生物可借助体内的磁铁矿（Fe3O4）颗粒感受磁场[18],后者则认为生物体内的隐花色素（cryptochrome,Cry）可受光激发产生自由基对反应,是理想的磁受体[19]。这两种假说均具有坚实的理论与实验基础,且目前科学家们更倾向于认同生物磁响应过程是这两种机制共同作用的结果[20]。此外,近期我国科学家发现了另一种潜在的磁受体[21],即铁硫簇蛋白IscA（iron-sulfur cluster assembly）,IscA也被认为是一种潜在的磁受体蛋白（magnetoreceptor protein,MagR）。该蛋白可与隐花色素结合形成复合物,体外实验证明该复合物不仅可感受磁场,且自身具有磁特性。同时,另一项侧重于体内生物学实验的研究也支持铁硫簇蛋白中的一种：IscA1具有磁感受功能,但后者的研究结果表明该蛋白可不依赖隐花色素单独调控神经活动[22]。【本研究切入点】目前,对于昆虫的磁生理响应多集中于中等磁场和强磁场,而对于弱磁场特别是地磁场和近零磁场环境下昆虫的生理变化研究较少[23]。褐飞虱（Nilaparvata lugens）是一种典型的夜间迁飞型害虫,而夜间迁飞的昆虫最可能利用地磁场作为其黑暗中的定向信号[24],暗示其具有磁响应的能力。利用超导量子磁强计结合普鲁士蓝染色观察发现,褐飞虱腹部含有铁磁性物质,可能为Fe3O4,且这种铁磁颗粒的含量与其翅型、性别和发育历期等有关[25]。研究表明,相较于地球磁场,近零磁场可影响褐飞虱与白背飞虱（Sogatella furcifera）的趋光性、飞行能力、卵及若虫历期、成虫寿命以及翅型比率等,且实时荧光定量PCR（RT-qPCR）结果显示,稻飞虱体内Cry1与Cry2表达量在两个磁场间存在显著差异[26,27],但进一步的分子验证仍是空白。【拟解决的关键问题】基于生物磁响应现象与磁感受机制的研究现状,以及前人针对稻飞虱对磁场的生理响应表型实验,选择褐飞虱雌雄成虫体内的磁响应关键基因,即NlCry1、NlCry2以及潜在磁受体蛋白基因NlMagR（即IscA）作为研究对象,通过RNA干扰（RNAi）技术分别敲减以上3种基因,并分别暴露于南京本地磁场（GMF：49 μT）和近零磁场（NZMF：1.8 μT）中,通过比较RNAi的褐飞虱雌雄成虫寿命的变化规律,探讨3种磁响应关键基因对磁场变化的响应及其对寿命的调控情况,为进一步明确磁受体及磁响应生物学通路打下基础。1 材料与方法
试验于2017年在南京农业大学完成。1.1 磁场发生装置的建立
采用自主研发的直流电亥姆霍兹线圈创造稳定的静态磁场环境（专利号：ZL201320004497.5）,以中心点为球心的直径30 cm的球形范围为该线圈可控制的有效磁场范围,本研究中所使用的几组线圈可控最小值范围为500—1 800 nT,因此取1.8 μT为本试验中近零磁场磁场强度。共设置两个稳定静态磁场,即模拟南京本地磁场强度的地磁场（GMF：49 μT）,以及抵消绝大部分地磁场以达到近零水平的近零磁场（NZMF：1.8 μT）。在磁场搭建调试、试验的开始、过程及结束阶段,使用磁通门计监测各磁场处理的强度,以保证控制环境试验中磁场强度设置的稳定性。1.2 供试昆虫与试验条件
供试褐飞虱采集自位于南京市的江苏省农业科学院试验稻田,饲养于室内种植的感虫水稻品种TN1上,温室环境为温度（26±1）℃,相对湿度80%,光周期14 L﹕10 D,磁场环境为南京本地地磁场。供试种群已于该环境内连续培养10代以上,且用于寿命分析的试虫均为发育整齐的短翅型雌雄成虫。1.3 主要试剂
总RNA提取试剂RNAiso Plus（TaKaRa）、反转录试剂盒PrimeScriptTM RT Reagent Kit with gDNA Eraser（TaKaRa）、凝胶回收试剂盒AxyPrepTM DNA Gel Extraction Kit（Axygen）、PCR试剂2×Fast Taq Master Mix（Gentbios）、零背景pTOPO-TA Simple克隆试剂盒（Aidlab）、感受态细胞Trans1-T1（TransGen Biotech）、质粒抽提与纯化试剂盒DNA Clean & ConcentratorTM -5（ZYMO RESEARCH）、体外转录试剂盒T7 RiboMAXTM Express RNAi System（Promega）、RT-qPCR试剂SYBR? Premix Ex TaqTM（TaKaRa）。引物合成及测序均由南京金斯瑞科技有限公司完成。
1.4 dsRNA的制备
根据前期研究[28,29],获得褐飞虱NlCry1（GenBank登录号：KM108578）、NlCry2（GenBank登录号：KM108579）、NlMagR（NlIscA）（GenBank登录号：KY026177）基因序列,经NCBI网站进行序列比对鉴定。应用Primer Premier 5.0软件设计3种基因的上下游克隆引物,同时设计绿色荧光蛋白基因GFP的克隆引物（表1,下划线为T7启动子）。其中阴性对照GFP的模板来自实验室保存的菌种。Table 1
表1
表1本试验所用引物及序列
Table 1
| 引物用途 Use of primers | 引物名称 Primer name | 引物序列 Primer sequence (5′-3′) |
|---|---|---|
| cDNA克隆 cDNA cloning | dsNlCry1-F | TAATACGACTCACTATAGGGTCAGACATGGGCTTCGATT |
| dsNlCry1-R | TAATACGACTCACTATAGGGCATTTGGTAAGTTAGCGGTGGA | |
| dsNlCry2-F | TAATACGACTCACTATAGGGTGTCAGCATCAATAAGTGGAGG | |
| dsNlCry2-R | TAATACGACTCACTATAGGGGCACACCAAACTTGTCGTC | |
| dsNlMagR-F | TAATACGACTCACTATAGGGAGAAAGGAAAGTTTGACGAAG | |
| dsNlMagR-R | TAATACGACTCACTATAGGGAGCCCTAAATATTAACATCGT | |
| dsGFP-F | TAATACGACTCACTATAGGGACGTAAACGGCCACAAGTTC | |
| dsGFP-R | TAATACGACTCACTATAGGGTGTTCTGCTGGTAGTGGTCG | |
| 实时荧光定量PCR RT-qPCR | qNlCry1-F | CAGACATGGGCTTCGATTTCA |
| qNlCry1-R | ACCAGCACTTTCTCCGTCAAAT | |
| qNlCry2-F | CGCATACTCTCTACAGACTTGAT | |
| qNlCry2-R | CACCGTCTGGAATTTGCGATAC | |
| qNlMagR-F | CGTTTAATACCTTCAAGAGCAGCAC | |
| qNlMagR-R | CCCTACTTTCAAGCCGATAGCAT | |
| qActin-F | CTTCTAAACGCCAACCACTCC | |
| qActin-R | TCACCCGAAATCACTCACGA | |
| q18S-F | TGTCTGCTTAATTGCGATAACGAAC | |
| q18S-R | CCTCAAACTTCCATCGGCTTG |
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取1.2中饲养的褐飞虱雌雄成虫各3只,同性别试虫冰浴中混合研磨匀浆后采用RNAiso Plus提取褐飞虱总RNA,其质量通过琼脂糖凝胶电泳进行检测,浓度通过Nanodrop2000（Thermo,USA）进行测定,随后按照反转录试剂盒PrimeScriptTM RT Reagent Kit的操作要求,获得可作为模板的cDNA序列。PCR反应体系为2×FastTaq PCR Master Mix 25 μL,10 μmol·L-1的上下游引物各1.5 μL,cDNA模板4 μL,ddH2O 18 μL,总反应体系为50 μL。PCR反应条件为95℃预变性3 min,95℃变性20 s,60℃复性20 s,72℃延伸30 s,30个循环,最后72℃延伸5 min。反应产物经琼脂糖凝胶电泳检测无误后,通过凝胶回收试剂盒AxyPrepTM DNA Gel Extraction Kit回收特定片段。
通过零背景pTOPO-TA Simple克隆试剂盒,将含有T7启动子的目的片段插入pTOPO-T质粒载体,转入大肠杆菌感受态细胞Trans1-T1,均匀涂布在含有1%青霉素的LB固体培养基上37℃倒置培养9 h,挑取单个菌落于1%青霉素的LB液体培养基中37℃摇菌5 h,取1 μL菌液进行PCR扩增,并送南京金斯瑞科技有限公司测序,测序结果通过MEGA软件进行比对。最终,按照DNA Clean & ConcentratorTM-5试剂盒的说明书对合格的菌液进行质粒抽提,随后按照体外转录试剂盒T7 RiboMAXTM Express RNAi System的操作说明合成dsRNA,退火及纯化后最终获得4种dsRNA,即dsNlCry1、dsNlCry2、dsNlMagR及dsGFP,调整浓度至4 000 ng·μL-1储存于-80℃备用。
1.5 RNAi
随机选取于1.2中饲养至初羽化1 d的短翅型雌雄成虫进行显微注射,以其胸部中足和后足间的外侧表皮处作为注射位点,并根据雌雄虫体形差异,确定雌虫注射量为75 nL,雄虫注射量为50 nL。共设3个处理组,即注射 dsNlCry1、dsNlCry2 和 dsNlMagR;以注射dsGFP 的试虫为阴性对照组。组内再分雌雄性别和不同磁场处理。因磁场空间有限,该试验分3批进行。注射后的试虫以单头单管的形式饲养于试管内种植的15日龄TN1水稻苗上,并分别置于南京本地磁场的GMF（49 μT）和近零磁场NZMF（1.8 μT）中,温湿度和光周期等环境条件同1.2。每日记录各处理试虫死亡情况。1.6 干扰效率检查
于RNAi处理后的24、48及72 h，随机取各处理位于GMF中的雌虫各9只，生物学重复设置3次，进行RNA抽提及基因RNAi后表达量情况测定。RT-qPCR采用20 μL体系，即SYBR? Premix Ex TaqTM 10 μL，上下游引物（10 μmol·L-1）各0.4 μL，ROX Reference Dey 0.4 μL，cDNA模板2 μL，ddH2O 6.8 μL。RT-qPCR反应采用两步法程序，即95℃预变性30 s，95℃变性5 s，60℃退火34 s，循环40次。扩增片段（引物列于表 1）经过测序和原序列比对，以β-actin和18S基因为双内参，以Comparative ΔΔCT法检测褐飞虱NlCry1、NlCry2及NlMagR的相对表达量。1.7 数据统计与分析
试验数据通过Excel 2010软件进行汇总整理,并采用SPSS16.0软件进行数据分析。其中,dsRNA的干扰效率通过独立样本T检验进行分析,敲减关键磁响应基因（NlCry1、NlCry2、NlMagR）、磁场变化（NZMF vs GMF）及其交互作用对褐飞虱雌雄成虫寿命的影响分别通过双因子方差分析（Two-way ANOVA）进行比较,并采用群体T检验进行处理间差异显著性比较（P<0.05为显著,P<0.01为极显著）。2 结果
2.1 dsRNA干扰效率
RT-qPCR结果显示,与注射dsGFP的阴性对照相比,注射dsNlCry1、dsNlCry2和dsNlMagR的雌成虫体内各对应基因1—3 d的相对表达量均显著或极显著降低（P<0.05或0.01）。与注射dsGFP的阴性对照相比,注射dsNlCry1 2 d后试虫体内NlCry1表达量显著降低了71%（P<0.05）,注射3 d后该基因表达量仍维持在较低水平,但差异不显著（图1-A）;注射dsNlCry2 1—2 d后试虫体内NlCry2表达量分别极显著降低了49%和54%（P<0.01,图1-B）;注射dsNlMagR 2—3 d后试虫体内NlMagR表达量分别显著降低了60%和53%（P<0.05,图1-C）。可见,显微注射dsRNA对3个磁响应关键的靶标基因具有相应的干扰效果。图1
新窗口打开|下载原图ZIP|生成PPT图1分别注射dsNlCry1、dsNlCry2和dsNlMagR后褐飞虱初羽化短翅型雌成虫各靶标RNA相对表达量
试虫均为初羽化短翅型雌成虫,置于B=49 μT的GMF中;*和**分别表示经T检验处理间差异达P<0.05和P<0.01的显著水平
Fig. 1Relative expression level of corresponding RNA of the newly emerged brachypterous female adults of N. lugens after respectively injected with dsNlCry1, dsNlCry2 and dsNlMagR
All of the sampled brown planthoppers are newly emerged brachypterous female adults and placed in the geomagnetic field that B=49 μT; * and ** mean significantly different between the dsRNA and the control dsGFP by the T test at P<0.05 and P<0.01, respectively
2.2 敲减NlCry1后褐飞虱雌雄成虫在正常磁场和近零磁场环境下的寿命
RNAi、磁场变化及两者之间的交互作用均未显著影响褐飞虱雌雄成虫的寿命（F≤2.87,P≥0.092,表2）。Table 2
表2
表2RNAi（dsNlCry1 vs. dsGFP）和磁场处理（GMF vs. NZMF）对褐飞虱雌雄成虫寿命影响的双因素方差分析
Table 2
| 影响因子 Impact factor | 雌虫寿命 Female longevity | 雄虫寿命 Male longevity | ||
|---|---|---|---|---|
| F值 F value | P值 P value | F值 F value | P值 P value | |
| RNAi | 2.87 | 0.092 | 0.90 | 0.34 |
| 磁场 Magnetic?field (MF) | 0.34 | 0.56 | 0.60 | 0.44 |
| RNAi×磁场 RNAi×MF | 0.54 | 0.47 | 1.88 | 0.17 |
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与注射dsGFP的对照相比,GMF下注射dsNlCry1的雌雄成虫寿命分别缩短了8.36%（图2-A）和20.26%（图2-B）,NZMF下注射dsNlCry1的雌雄成虫寿命分别延长了11.97%（图2-A）和7.23%（图2-B）;与GMF相比,NZMF下注射dsNlCry1的雌成虫寿命缩短了6.70%（图2-A）,而雄成虫寿命延长了5.00%（图2-B）。但以上RNAi处理以及磁场处理间差异均不显著（P>0.05,图2）。
图2
新窗口打开|下载原图ZIP|生成PPT图2注射dsNlCry1后正常磁场和近零磁场下褐飞虱雌成虫（A）和雄成虫（B）寿命
相同小写字母和相同大写字母分别表示同一磁场同一性别褐飞虱成虫寿命在dsNlCry1与dsGFP处理之间,以及同一性别同一RNAi干扰处理下GMF和NZMF处理之间经T检验差异不显著（P>0.05）
Fig. 2The longevity of female (A) and male (B) adults of N. lugens after dsNlCry1 injection under geomagnetic field (GMF) and near-zero magnetic field (NZMF)
The same lowercase and uppercase letters indicate no significant difference between the treatments of dsNlCry1 and dsGFP for the same sex N. lugens under same magnetic field of GMF or NZMF, and between the treatments of GMF and NZMF for same sex N. lugens injected of dsNlCry1 or dsGFP by the T test (P>0.05), respectively
2.3 敲减NlCry2后褐飞虱雌雄成虫在正常磁场和近零磁场环境下的寿命
磁场变化显著影响了褐飞虱雄成虫寿命（F=4.14,P=0.044<0.05）,且RNAi和磁场变化之间的交互作用还显著影响了雌成虫寿命（F=4.45,P=0.037<0.05）,并极显著地影响了雄成虫寿命（F=7.08,P=0.009<0.01,表3）。与注射dsGFP的对照相比,GMF下注射dsNlCry2的雌成虫寿命缩短了2.59%,而NZMF下注射dsNlCry2的雌成虫寿命延长了13.07%,但差异均不显著（P>0.05,图3-A）;GMF下注射dsNlCry2的雄成虫寿命显著缩短了25.41%（P<0.05）,而在NZMF下注射dsNlCry2的雄成虫寿命缩短了10.73%,但差异不显著（P>0.05,图3-B）。与GMF相比,NZMF下注射dsNlCry2的褐飞虱雌雄成虫寿命分别显著延长了27.28%和50.04%（P<0.05,图3）。Table 3
表3
表3RNAi（dsNlCry2 vs. dsGFP）和磁场处理（GMF vs. NZMF）对褐飞虱雌雄成虫寿命影响的双因素方差分析
Table 3
| 影响因子 Impact factor | 雌虫寿命 Female longevity | 雄虫寿命 Male longevity | ||
|---|---|---|---|---|
| F值 F value | P值 P value | F值 F value | P值 P value | |
| RNAi | 2.33 | 0.13 | 1.64 | 0.20 |
| 磁场 Magnetic?field (MF) | 0.70 | 0.40 | 4.14 | 0.044* |
| RNAi×磁场 RNAi×MF | 4.45 | 0.037* | 7.08 | 0.009** |
新窗口打开|下载CSV
图3
新窗口打开|下载原图ZIP|生成PPT图3注射dsNlCry2后正常磁场和近零磁场下褐飞虱雌成虫（A）和雄成虫（B）寿命
不同小写字母和不同大写字母分别表示同一磁场同一性别褐飞虱成虫寿命在dsNlCry2与dsGFP处理之间,以及同一性别同一RNAi干扰处理下GMF和NZMF处理之间经T检验差异显著（P<0.05）
Fig. 3The longevity of female (A) and male (B) adults of N. lugens after dsNlCry2 injection under geomagnetic field (GMF) and near-zero magnetic field (NZMF)
Different lowercase and uppercase letters indicate significantly different between the treatments of dsNlCry2 and dsGFP for the same sex N. lugens under same magnetic field of GMF or NZMF, and between the treatments of GMF and NZMF for same sex N. lugens injected of dsNlCry2 or dsGFP by the T test (P<0.05), respectively
2.4 敲减NlMagR后褐飞虱雌雄成虫在正常磁场和近零磁场环境下的寿命
RNAi及其与磁场变化之间的交互作用均未显著影响褐飞虱雌雄成虫寿命（F≤2.59,P≥0.11）,敲减NlMagR仅在NZMF下显著影响了雌成虫寿命（F=3.97,P=0.048<0.05,表4）。与注射dsGFP的对照相比,GMF下注射dsNlMagR的雌成虫寿命缩短6.69%,NZMF下则显著缩短16.48%（P<0.05,图4-A）;GMF下注射dsNlMagR的雄成虫寿命延长9.32%,而NZMF下则缩短12.98%,但二者差异并不显著（P>0.05,图4-B）;与GMF相比,NZMF下注射dsNlMagR的褐飞虱雌雄虫寿命分别缩短了12.16%和12.68%,但差异并未达到显著水平（P>0.05,图4）。Table 4
表4
表4RNAi（dsNlMagR vs. dsGFP）和磁场处理（GMF vs. NZMF）对褐飞虱雌雄虫寿命影响的双因素方差分析
Table 4
| 影响因子 Impact factor | 雌虫寿命 Female longevity | 雄虫寿命 Male longevity | ||
|---|---|---|---|---|
| F值 F value | P值 P value | F值 F value | P值 P value | |
| RNAi | 3.97 | 0.048* | 0.11 | 0.74 |
| 磁场 Magnetic?field (MF) | 1.33 | 0.25 | 0.08 | 0.78 |
| RNAi×磁场 RNAi×MF | 0.68 | 0.41 | 2.59 | 0.11 |
新窗口打开|下载CSV
图4
新窗口打开|下载原图ZIP|生成PPT图4注射dsNlMagR后正常磁场和近零磁场下褐飞虱雌成虫（A）和雄成虫（B）寿命
不同小写字母和不同大写字母分别表示同一磁场同一性别褐飞虱成虫寿命在dsNlMagR与dsGFP处理之间,以及同一性别同一RNAi干扰处理下GMF和NZMF处理之间经T检验差异显著（P<0.05）
Fig. 4The longevity of female (A) and male (B) adults of N. lugens after dsNlMagR injection under geomagnetic field (GMF) and near-zero magnetic field (NZMF)
Different lowercase and uppercase letters indicate significantly different between the treatments of dsNlMagR and dsGFP for the same sex N. lugens under same magnetic field of GMF or NZMF, and between the treatments of GMF and NZMF for same sex N. lugens injected of dsNlMagR or dsGFP by the T test (P<0.05), respectively
3 讨论
近年来,大量研究表明隐花色素（Cry）可作为潜在磁受体参与生物的磁场响应过程。昆虫中的果蝇只含有一种隐花色素,可作为研究该蛋白响应磁场变化的模式生物。有关行为学实验表明,全光照下野生型果蝇可对磁场作出显著反应,而隐花色素缺陷的果蝇突变体则对磁场无响应,有趣的是,当屏蔽蓝紫光（即隐花色素可接收的光谱）后,即使是野生型果蝇也不再响应磁场的变化,证实了隐花色素的存在是果蝇进行磁响应活动的必要条件,也为基于隐花色素的磁敏系统提供了首个遗传学证据[30]。随后,在有关昼夜节律的研究中发现,蓝光光谱下磁场可增强隐花色素对果蝇生物钟节律的“减缓”现象,而隐花色素突变体果蝇中则未观察到这一现象[31]。此外,另有研究发现,果蝇的负趋地性会受到磁场的干扰,而这种“干扰”又受隐花色素的调节,具体而言,破坏隐花色素的羧基端后,磁场对果蝇负趋地性的影响消失,证明隐花色素在这一信号通路中发挥着重要的作用[32]。近期一项针对磁场对稻飞虱生物学效应的研究表明,与GMF相比,NZMF使白背飞虱若虫历期延长,并显著缩短了长翅未交配白背飞虱成虫寿命;且RT-qPCR结果显示,NZMF下5龄若虫体内的Cry1和Cry2均上调表达[33]。本研究中,注射dsNlCry2后,相比GMF,NZMF中褐飞虱雌雄成虫寿命均显著延长,而作为阴性对照的dsGFP处理组雌雄虫寿命在磁场间均无显著差异,且双因素方差分析显示,磁场与干扰互作对褐飞虱成虫寿命有极显著影响,表明该基因可对磁场变化产生敏感响应。此外,磁场与敲减NlCry2的交互作用对不同性别的褐飞虱成虫寿命的影响不同,具体表现为注射dsNlCry1后,NZMF下褐飞虱雌雄成虫寿命均呈现延长趋势;注射dsNlMagR后,NZMF下褐飞虱雌雄成虫寿命均出现缩短的趋势;而注射dsNlCry2后,NZMF下雌成虫寿命延长,雄成虫寿命缩短,即表现“性二型性”。综上所述,NlCry2参与了褐飞虱成虫的寿命调控,且该过程受到磁场影响,符合隐花色素可响应磁场变化的假说。本研究中,干扰NlCry2后褐飞虱的成虫寿命在不同磁场处理间存在显著差异,而干扰NlCry1后褐飞虱的成虫寿命在磁场处理间无显著差异,可见不同磁场环境下两种隐花色素调控褐飞虱成虫寿命的功能不同。隐花色素主要在动物的生物钟调控方面发挥重要作用,可分为两类,即光敏的Ⅰ型隐花色素和非光敏的Ⅱ型隐花色素;其中,Ⅰ型为蓝光受体,在生物钟系统中起着感光作用,而Ⅱ型为昼夜节律调节过程中的反馈回路抑制子[34]。褐飞虱的两种隐花色素基因NlCry1与NlCry2分别属于动物Ⅰ型与动物Ⅱ型隐花色素。有研究表明,NlCry1与NlCry2在褐飞虱的发育生理中扮演着不同的角色,前者在褐飞虱的迁徙行为中发挥作用,而后者对昼夜节律的适应显示出更明显的动态变化[29]。可见NlCry2更有可能作为动物生物钟的核心组件。而生物钟又可与神经内分泌系统协同作用[35,36,37],二者可能间接调控昆虫体内激素稳态,如蜕皮激素、保幼激素的分泌等[38,39,40]。因此,NlCry2可能作为关联磁场效应与激素信号传导的“桥梁”,进而在褐飞虱的生物钟系统中发挥潜在作用。另一方面,NlCry1或与稻飞虱的趋光行为有关。WAN等研究表明,NZMF显著提高了初羽化2日龄的白背飞虱内的正趋光性;且RT-qPCR结果显示,该时段内其体内NlCry1上调表达,进而推测NZMF提高白背飞虱正趋光性或与NlCry1羧基端多效性有关[27]。因此,作为Ⅰ型隐花色素,NlCry1可能更多发挥感光受体作用,或参与调节磁场影响下的起飞或趋光等行为,而不参与磁场对褐飞虱寿命的调控。
铁硫簇蛋白IscA属于铁硫蛋白亚家族hesB,是一类高度保守的蛋白。近期,QIN等研究发现该蛋白在离体条件下具有磁性,推测可能是一种全新的磁受体蛋白[21,22]。QIN等[21]以离体实验为主,认为铁硫簇蛋白与隐花色素结合,前者感磁,后者感光,协同发挥“生物指南针”功能;而LONG等[22]以线虫为研究对象,认为铁硫簇蛋白不需与隐花色素结合便可发挥磁受体功能。但该理论提出后受到质疑,且缺乏活体验证,至今仍无后续相关报道。本研究中,NZMF下干扰NlMagR的褐飞虱雌虫寿命显著短于对照处理,而雄虫中虽有相似趋势,却未达显著水平,推测NlMagR或对磁场存在一定的感知,且它的缺失可缩短雌虫寿命,进而影响其生长发育。铁硫簇蛋白是一类重要的线粒体内蛋白,MagR（IscA）对于维持线粒体稳定以及调控呼吸能量代谢具有重要生物学意义。有研究表明,缺失该基因会导致铁元素在酵母菌线粒体内累积,进而使其无法正常生长[41,42,43]。此外MagR（IscA）也是线粒体铁硫蛋白成熟所必需的组装蛋白,敲除斑马鱼体内的IscA会导致其血红蛋白缺失,从而造成重度贫血[44]。最近有关该基因在人类疾病方面的研究也表明,IscA的异常表达会导致某些疾病的发生,如脑白质病变与神经退行性疾病[45]以及槭糖尿病[46]。本试验中,GMF条件下干扰NlMagR后,相较于dsGFP处理,褐飞虱雌成虫寿命缩短但不显著,但NZMF条件下干扰该基因后的试虫寿命进一步缩短达并到显著水平,这可能暗示了：（1）NlMagR（即铁硫簇蛋白）的缺失对褐飞虱雌成虫有消极影响,原因可能是上文提到的线粒体或铁硫蛋白受损等;（2）NZMF“放大”了NlMagR缺失带来的消极影响。据此推测NlMagR可以感受磁场的变化,并在褐飞虱雌成虫的寿命这一体征中有所体现,符合“生物磁受体”的理论要求。
4 结论
对于迁飞型害虫褐飞虱而言,其体内NlCry2对磁场变化存在响应,表现为敲减该基因与磁场变化的互作可对褐飞虱雌雄虫寿命产生显著影响,且具有“性二型性”;NlMagR（IscA）也可对磁场产生一定响应,但该响应只存在于雌虫中,雄虫的影响不明显;而以寿命为指标不能判定NlCry1对磁场变化是否存在响应,该基因或与褐飞虱寿命调节无关。研究结果可为进一步明确生物磁受体以及磁响应生物学通路提供有力的理论支撑。参考文献 原文顺序
文献年度倒序
文中引用次数倒序
被引期刊影响因子
DOI:10.3969/j.issn.0253-9608.2013.03.007URLMagsci [本文引用: 1]
<p>两千多年前，人类发明了指南针用于辨别方向；而如今科学家们发现，很多动物也能利用体内的生物指南针<br />感应磁场。笔者从磁场的产生入手，详细介绍了地磁场的性质和生物磁感应现象的产生。候鸟是最早被注意到能利<br />用磁场导航，且目前已获得最多磁生物学研究结果的一类高等生物。大量的行为学实验证明，候鸟在长距离迁徙的<br />过程中主要靠对地磁场的感应来确定方向，它们的大脑能记录下每一个特殊地点的磁特征，并据此找出到达各个目<br />的地的飞行路线。不仅如此，部分鸟类的磁导航还有一定程度的蓝光依赖性、会受到异常磁场的干扰，这种现象可<br />以用生物磁受体的磁铁矿的感应假说和化学感受假说来解释。很多生物，包括人类也都有与候鸟类似的磁受体，有<br />些动物能用它们感受磁场并以此作为视觉和听觉的辅助，而其他生物的磁感应能力也许已经在进化的某个阶段被别<br />的功能替代或是直接消失了。目前人们对于生物磁现象的研究才刚刚起步，还有很多未知的谜团等待我们去揭开，<br />希望在不久的将来能看到更多令人惊讶的实验结果。</p>
DOI:10.3969/j.issn.0253-9608.2013.03.007URLMagsci [本文引用: 1]
<p>两千多年前，人类发明了指南针用于辨别方向；而如今科学家们发现，很多动物也能利用体内的生物指南针<br />感应磁场。笔者从磁场的产生入手，详细介绍了地磁场的性质和生物磁感应现象的产生。候鸟是最早被注意到能利<br />用磁场导航，且目前已获得最多磁生物学研究结果的一类高等生物。大量的行为学实验证明，候鸟在长距离迁徙的<br />过程中主要靠对地磁场的感应来确定方向，它们的大脑能记录下每一个特殊地点的磁特征，并据此找出到达各个目<br />的地的飞行路线。不仅如此，部分鸟类的磁导航还有一定程度的蓝光依赖性、会受到异常磁场的干扰，这种现象可<br />以用生物磁受体的磁铁矿的感应假说和化学感受假说来解释。很多生物，包括人类也都有与候鸟类似的磁受体，有<br />些动物能用它们感受磁场并以此作为视觉和听觉的辅助，而其他生物的磁感应能力也许已经在进化的某个阶段被别<br />的功能替代或是直接消失了。目前人们对于生物磁现象的研究才刚刚起步，还有很多未知的谜团等待我们去揭开，<br />希望在不久的将来能看到更多令人惊讶的实验结果。</p>
DOI:10.1016/j.micron.2004.12.009URL [本文引用: 1]
DOI:10.3724/SP.J.1206.2011.00597URLMagsci [本文引用: 1]
Here, we review the progresses of researches on the biological effects of the hypomagnetic field (HMF) and propose that a magnetic field with magnetic induction of "0 < |B| ≤ 5 μT" is defined as a hypomagnetic field (HMF)．Interplanetary space is a natural hypomagnetic field．The astronauts, enabled by space sciences thriving in the recent decades, are spending longer time in the hypomagnetic outer space, e.g．landing on the moon and heading to mars. The effects of HMF on many aspects of biological processes, especially the adverse impacts on the functions of the central nervous system, remind us that the astronauts would suffer from potential risks due to the HMF exposure．Unfolding the mechanism of the biological responses to the HMF is the fundament for developing the counteractions of the adverse space environmental factors, and has recently become a hot topic in the field of space life sciences. Refining the concept of HMF and standardizing the HMF simulation systems for biological experiments will benefit the comparability of the HMF effects as described, and improve our understanding of how HMF influences homeostasis, cell signaling pathways and cognitive behaviors, and to what extent HMF contribute to such disturbances.
DOI:10.3724/SP.J.1206.2011.00597URLMagsci [本文引用: 1]
Here, we review the progresses of researches on the biological effects of the hypomagnetic field (HMF) and propose that a magnetic field with magnetic induction of "0 < |B| ≤ 5 μT" is defined as a hypomagnetic field (HMF)．Interplanetary space is a natural hypomagnetic field．The astronauts, enabled by space sciences thriving in the recent decades, are spending longer time in the hypomagnetic outer space, e.g．landing on the moon and heading to mars. The effects of HMF on many aspects of biological processes, especially the adverse impacts on the functions of the central nervous system, remind us that the astronauts would suffer from potential risks due to the HMF exposure．Unfolding the mechanism of the biological responses to the HMF is the fundament for developing the counteractions of the adverse space environmental factors, and has recently become a hot topic in the field of space life sciences. Refining the concept of HMF and standardizing the HMF simulation systems for biological experiments will benefit the comparability of the HMF effects as described, and improve our understanding of how HMF influences homeostasis, cell signaling pathways and cognitive behaviors, and to what extent HMF contribute to such disturbances.
URL [本文引用: 1]
地球上的生物每时每刻都受到地球磁场的影响.本文从介绍磁场及地球磁场入手,详细阐述了生物对地球磁场的磁响应现象.从候鸟到厌氧细菌,大量生物已被证实可对磁场产生生理和行为响应,即表现为磁场影响生物的生理发育,且提供地磁信息作为"罗盘"与"地图",指导动物远距离迁徙或短距离扩散等.关于生物磁响应的机制,目前有两种假说得到广泛认可,即磁颗粒介导的磁受体假说和依赖光并基于自由基对的磁响应假说,而近期由中国科学家提出的磁蛋白生物指南针模型更是引起了广泛关注,但至今尚缺乏生物活体验证.目前,全球变化背景下的地磁场变化面临加剧风险,而国际上(尤其是国内)对生物磁响应现象的研究才刚刚起步,相关研究亟待深入开展,以便为应对地磁场变化提供基于生物磁响应的科学依据.
URL [本文引用: 1]
地球上的生物每时每刻都受到地球磁场的影响.本文从介绍磁场及地球磁场入手,详细阐述了生物对地球磁场的磁响应现象.从候鸟到厌氧细菌,大量生物已被证实可对磁场产生生理和行为响应,即表现为磁场影响生物的生理发育,且提供地磁信息作为"罗盘"与"地图",指导动物远距离迁徙或短距离扩散等.关于生物磁响应的机制,目前有两种假说得到广泛认可,即磁颗粒介导的磁受体假说和依赖光并基于自由基对的磁响应假说,而近期由中国科学家提出的磁蛋白生物指南针模型更是引起了广泛关注,但至今尚缺乏生物活体验证.目前,全球变化背景下的地磁场变化面临加剧风险,而国际上(尤其是国内)对生物磁响应现象的研究才刚刚起步,相关研究亟待深入开展,以便为应对地磁场变化提供基于生物磁响应的科学依据.
DOI:10.1242/jeb.001313URLPMID:17951410 [本文引用: 1]
Abstract Diverse animals detect the Earth's magnetic field and use it as a cue in orientation and navigation. Most research on magnetoreception has focused on the directional or ;compass' information that can be extracted from the Earth's field. Because the field varies predictably across the surface of the globe, however, it also provides a potential source of positional or 'map' information, which some animals use to steer themselves along migratory pathways or to navigate toward specific target areas. The use of magnetic positional information has been demonstrated in several diverse animals including sea turtles, spiny lobsters, newts and birds, suggesting that such systems are phylogenetically widespread and can function over a wide range of spatial scales. These ;magnetic maps' have not yet been fully characterized. They may be organized in several fundamentally different ways, some of which bear little resemblance to human maps, and they may also be used in conjunction with unconventional navigational strategies.
DOI:10.1002/1522-2586(200007)12:1<2::AID-JMRI2>3.0.CO;2-VURLPMID:10931560 [本文引用: 1]
Abstract Issues associated with the exposure of patients to strong, static magnetic fields during magnetic resonance imaging (MRI) are reviewed and discussed. The history of human exposure to magnetic fields is reviewed, and the contradictory nature of the literature regarding effects on human health is described. In the absence of ferromagnetic foreign bodies, there is no replicated scientific study showing a health hazard associated with magnetic field exposure and no evidence for hazards associated with cumulative exposure to these fields. The very high degree of patient safety in strong magnetic fields is attributed to the small value of the magnetic susceptibility of human tissues and to the lack of ferromagnetic components in these tissues. The wide range of susceptibility values between magnetic materials and human tissues is shown to lead to qualitatively differing behaviors of these materials when they are exposed to magnetic fields. Mathematical expressions are provided for the calculation of forces and torques. J. Magn. Reson. Imaging 2000;12:2 19. 2000 Wiley-Liss, Inc.
DOI:10.1385/CBB:39:2:163URLPMID:14515021 [本文引用: 1]
There is substantial evidence indicating that moderate-intensity static magnetic fields (SMF) are capable of influencing a number of biological systems, particularly those whose function is closely linked to the properties of membrane channels. Most of the reported moderate SMF effects may be explained on the basis of alterations in membrane calcium ion flux. The mechanism suggested to explain these effects is based on the diamagnetic anisitropic properties of membrane phospholipids. It is proposed that reorientation of these molecules during moderate SMF exposure will result in the deformation of imbedded ion channels, thereby altering their activation kinetics. Channel inactivation would not be expected to be influenced by these fields because this mechanism is not located within the intramembraneous portion of the channel. Patch-clamp studies of calcium channels have provided support for this hypothesis, as well as demonstrating a temperature dependency that is understandable on the basis of the membrane thermotropic phase transition. Additional studies have demonstrated that sodium channels are similarly affected by SMFs, although to a lesser degree. These findings support the view that moderate SMF effects on biological membranes represent a general phenomenon, with some channels being more susceptible than others to membrane deformation.
DOI:10.3321/j.issn:0023-074X.2003.19.008URLMagsci [本文引用: 1]
自然地磁场中孵化的一日龄小鸡(对照组)一次性味觉回避学习行为(OTPAT)的记忆保持曲线与已公认的三阶段模型基本相一致, 第20分钟和第60分钟为两个低谷, 短时和中间记忆时段对红色小珠的回避率(AR<SUB>R</SUB>)平均为68.4%, 长时记忆时段的AR<SUB>R</SUB>平均为74.8%. 亚磁空间中孵化的一日龄小鸡(实验组)OTPAT的记忆保持曲线存在显著的时间效应, 第25分钟和第50分钟为两个低谷, 短时和中间记忆与对照组相似, AR<SUB>R</SUB>平均为74.1%; 但长时记忆出现明显的振荡现象, AR<SUB>R</SUB>比对照组平均下降25.3%, 变差系数增大1.3倍, 即其记忆能力和稳定性都显著变差. 这表明, 近似地磁场消除的亚磁空间对鸡胚脑功能发育具有不可忽视的负面影响.
DOI:10.3321/j.issn:0023-074X.2003.19.008URLMagsci [本文引用: 1]
自然地磁场中孵化的一日龄小鸡(对照组)一次性味觉回避学习行为(OTPAT)的记忆保持曲线与已公认的三阶段模型基本相一致, 第20分钟和第60分钟为两个低谷, 短时和中间记忆时段对红色小珠的回避率(AR<SUB>R</SUB>)平均为68.4%, 长时记忆时段的AR<SUB>R</SUB>平均为74.8%. 亚磁空间中孵化的一日龄小鸡(实验组)OTPAT的记忆保持曲线存在显著的时间效应, 第25分钟和第50分钟为两个低谷, 短时和中间记忆与对照组相似, AR<SUB>R</SUB>平均为74.1%; 但长时记忆出现明显的振荡现象, AR<SUB>R</SUB>比对照组平均下降25.3%, 变差系数增大1.3倍, 即其记忆能力和稳定性都显著变差. 这表明, 近似地磁场消除的亚磁空间对鸡胚脑功能发育具有不可忽视的负面影响.
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DOI:10.1002/bem.20056URLPMID:15672364 [本文引用: 1]
We have recently observed that a single exposure of mice to a magnetically shielded environment can attenuate opioid induced analgesia. Here, we report the effect of repeated exposures to the same magnetically shielded environment. Adult male Swiss CD-1 mice were placed in a Mu-metalTM lined box or an opaque PlexiglasTM box (sham condition) for 1 h per day for 10 consecutive days. Nociception was measured as the latency time to a foot lift/lick in response to an aversive thermal stimulus (hotplate analgesiometer, 50 1 C) before and immediately after exposure. Multiple experiments were conducted in which thermal latency was tested on each of the 10 days or on days 1, 5, and 10, with some utilizing post-exposure testing only. It was shown that mice can detect and will respond to the repeated absence of the ambient magnetic field, with a maximum analgesic response occurring over days 4-6 of exposure and returning to baseline thereafter. The effect was robust, independent of pre-exposure and intermittent testing, and seems to be opioid related, since the results obtained on day 5 were similar to those from a 5 mg/kg dose of morphine and were abolished with the opioid antagonist, naloxone. Bioelectromagnetics 26:109-117, 2005. ? 2005 Wiley-Liss, Inc.
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DOI:10.1038/261411a0URLPMID:934271 [本文引用: 1]
http://www.nature.com/doifinder/10.1038/261411a0
DOI:10.3109/15368371003627290URLPMID:20230271 [本文引用: 1]
It was shown that the 250-fold screening of the geomagnetic field (GMF) (090008zero090009 magnetic field with an induction of 0.2 0204T) affects early embryogenesis and the reproduction capacity of mice in vivo. Pregnant NMRI mice at the zygote stage placed in this 090008zero090009 magnetic field (MF) lost the ability to bear offspring babies although their embryos developed up to the blastocyst stage without any visible deviations from the norm. The abortion of development in the 090008zero090009 MF occurred after the exit of the blastocysts from the zona pellicida and invasion into the uterus during implantation. Histological analysis indicates that possible reasons of the abnormalities of postimplantation development are a decrease in the proliferative activity of embryonic cells and the impairment of the interaction between the trophoblast and endometrium, which finally results in the resorption of embryos in the uterus.
DOI:10.1002/bem.20699URLPMID:21853450 [本文引用: 1]
<P>The hypogeomagnetic field (HGMF; magnetic fields <20065nT) is one of the fundamental environmental factors of space. However, the effect of HGMF exposure on living systems remains unclear. In this article, we examine the biological effects of HGMF on the embryonic development of Xenopus laevis (African clawed frog). A decrease in horizontal third cleavage furrows and abnormal morphogenesis were observed in Xenopus embryos growing in the HGMF. HGMF exposure at the two-cell stage, but no later than the four-cell stage, is enough to alter the third cleavage geometry pattern. Immunofluorescent staining for -tubulin showed reorientation of the spindle of four-cell stage blastomeres. These results indicate that a brief (2-h) exposure to HGMF is sufficient to interfere with the development of Xenopus embryos at cleavage stages. Also, the mitotic spindle could be an early sensor to the deprivation of the geomagnetic field, which provides a clue to the molecular mechanism underlying the morphological and other changes observed in the developing and/or developed embryos. Bioelectromagnetics 33:238–246, 2012. 08 2011 Wiley Periodicals, Inc.</P>
DOI:10.3109/15368370903167246URLPMID:20001705 [本文引用: 1]
In our previous works, we reported that compensation of the geomagnetic field to a level less than 0.4 0204T (090008zero magnetic field,090009 or ZMF) affected human cognitive processes. ZMF exposure increased the number of errors and the task processing time by 2.4% in average. However, in the array of the magnetic effects calculated from the experimental data, some readings have been found to deviate from the mean magnetic effect by more than three standard deviations. This finding could give rise to doubt as to whether the magnetic effect observed was a mere sequence of the presence of such unlikely data values. In the present work we examine the results of the unlikely data elimination and show that the corrected magnetic effect in tested humans remains statistically significant, though at a reduced magnitude 1.5%.
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DOI:10.1134/S0362119716070057URLPMID:25087408 [本文引用: 1]
The impact of attenuated magnetic field ( F) on human health is a burning issue of present-day cosmonautics. A series of experiments with animals exposed to attenuated MF revealed violent disorders in the development of the cardiovascular system. The purpose of this study was to estimate the effects of hypomagnetic environment (HME) on capillary blood circulation, blood pressure (BP), and heart rate (HR) in healthy humans. Participants ( n = 34) were 24 men and 10 women without cardiovascular symptoms. The mean age was 43.3 15.4 years. Thirteen participants, eight men and five women, were randomly selected for a repeated investigation under natural conditions (sham exposure). The mean age in this group was 47.9 18 years. Cardiac rhythm and heart rate were recorded using an Astrocard cardiac monitor (Russia). BP was measured by means of a Tonocard automatic blood pressure monitor (Russia). Capillary circulation was determined using a digital capillaroscope (Russia) with a high-speed CMOS camera (100 frames/s). The duration of HME exposure was 60 min. It has been demonstrated that HME increases capillary circulation rate by 22.4% in healthy humans without cardiovascular symptoms as compared to the records made under natural conditions. There was a significant HR reduction by the end of HME exposure as compared to the measurements taken at the beginning. At the end of the exposure, diastolic BP dropped considerably relative to mid-exposure values; on the contrary, systolic BP significantly increased by the end.
DOI:10.1098/rsif.2015.0499URLPMID:26333810 [本文引用: 1]
Behavioural studies underpin the weight of experimental evidence for the existence of a magnetic sense in animals. In contrast, studies aimed at understanding the mechanistic basis of magnetoreception by determining the anatomical location, structure and function of sensory cells have been inconclusive. In this review, studies attempting to demonstrate the existence of a magnetoreceptor based on the principles of the magnetite hypothesis are examined. Specific attention is given to the range of techniques, and main animal model systems that have been used in the search for magnetite particulates. Anatomical location/cell rarity and composition are identified as two key obstacles that must be addressed in order to make progress in locating and characterizing a magnetite-based magnetoreceptor cell. Avenues for further study are suggested, including the need for novel experimental, correlative, multimodal and multidisciplinary approaches. The aim of this review is to inspire new efforts towards understanding the cellular basis of magnetoreception in animals, which will in turn inform a new era of behavioural research based on first principles.
DOI:10.1016/S0006-3495(00)76629-XURL [本文引用: 1]
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DOI:10.1038/nmat4484URLPMID:26569474 [本文引用: 3]
Abstract The notion that animals can detect the Earth's magnetic field was once ridiculed, but is now well established. Yet the biological nature of such magnetosensing phenomenon remains unknown. Here, we report a putative magnetic receptor (Drosophila CG8198, here named MagR) and a multimeric magnetosensing rod-like protein complex, identified by theoretical postulation and genome-wide screening, and validated with cellular, biochemical, structural and biophysical methods. The magnetosensing complex consists of the identified putative magnetoreceptor and known magnetoreception-related photoreceptor cryptochromes (Cry), has the attributes of both Cry- and iron-based systems, and exhibits spontaneous alignment in magnetic fields, including that of the Earth. Such a protein complex may form the basis of magnetoreception in animals, and may lead to applications across multiple fields.
DOI:10.1007/s11434-015-0902-0URLPMID:4692962 [本文引用: 3]
Current neuromodulation techniques such as optogenetics and deep-brain stimulation are transforming basic and translational neuroscience. These two neuromodulation approaches are, however, invasive since surgical implantation of an optical fiber or wire electrode is required. Here, we have invented a non-invasive magnetogenetics that combines the genetic targeting of a magnetoreceptor with remote magnetic stimulation. The non-invasive activation of neurons was achieved by neuronal expression of an exogenous magnetoreceptor, an iron-sulfur cluster assembly protein 1 (Isca1). In HEK-293 cells and cultured hippocampal neurons expressing this magnetoreceptor, application of an external magnetic field resulted in membrane depolarization and calcium influx in a reproducible and reversible manner, as indicated by the ultrasensitive fluorescent calcium indicator GCaMP6s. Moreover, the magnetogenetic control of neuronal activity might be dependent on the direction of the magnetic field and exhibits on-response and off-response patterns for the external magnetic field applied. The activation of this magnetoreceptor can depolarize neurons and elicit trains of action potentials, which can be triggered repetitively with a remote magnetic field in whole-cell patch-clamp recording. In transgenic Caenorhabditis elegans expressing this magnetoreceptor in myo-3-specific muscle cells or mec-4-specific neurons, application of the external magnetic field triggered muscle contraction and withdrawal behavior of the worms, indicative of magnet-dependent activation of muscle cells and touch receptor neurons, respectively. The advantages of magnetogenetics over optogenetics are its exclusive non-invasive, deep penetration, long-term continuous dosing, unlimited accessibility, spatial uniformity and relative safety. Like optogenetics that has gone through decade-long improvements, magnetogenetics, with continuous modification and maturation, will reshape the current landscape of neuromodulation toolboxes and will have a broad range of applications to basic and translational neuroscience as well as other biological sciences. We envision a new age of magnetogenetics is coming.
DOI:10.1002/bem.20290URLPMID:17016848 [本文引用: 1]
Previous studies found that elimination of the geomagnetic field (GMF) interferes with the normal brain functions, but the underlying mechanism remains unknown. The present study examined the effects of long-term exposures to a near-zero magnetic environment on the noradrenergic activities in the brainstem of golden hamsters. Both the content of norepinephrine (NE) and the density of NE-immunopositive neurons in the tissue decreased significantly after the treatment, and the effects could be progressive with time. These variations may substantially contribute to behavioral and mood disorders reported in other studies when animals are shielded from the GMF. Bioelectromagnetics 2006 Wiley-Liss, Inc.
DOI:10.1146/annurev-ento-120709-144820URL [本文引用: 1]
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DOI:10.1038/srep18771URLPMID:4700427 [本文引用: 1]
Biogenic magnetic particles have been detected in some migratory insects, which implies the basis of magnetoreception mechanism for orientation and navigation. Here, the biogenic magnetic particles in the migratory brown planthopper (BPH),Nilaparvata lugenswere qualitatively measured by SQUID magnetometry, and their characteristics were further determined by Prussian Blue staining, electron microscopy and energy dispersive x-ray spectroscopy. The results indicate that there were remarkable magnetic materials in the abdomens and not in the head or thorax of the 3rd–5thinstar nymphs, and in macropterous and brachypterous female and male adults of BPH. The size of magnetic particles was shown to be between 50–45065nm with a shape factor estimate of between 0.8–1.0 for all the tested BPHs. Moreover, the amount of magnetic particles was associated with the developmental stage (the 3rd–5thinstar), wing form (macropterous vs. brachypterous) and sex. The macropterous female adults had the largest amount of magnetic particles. Although the existence of magnetic particles in the abdomens of BPH provides sound basis for the assumption of magnetic orientation, further behavioral studies and complementary physical characterization experiments should be conducted to determine whether the orientation behavior of BPH is associated with the magnetic particles detected in this study.
DOI:10.1016/j.jinsphys.2014.06.016URLPMID:24995837 [本文引用: 1]
Magnetic fields markedly affect the growth and development of many species of organisms potentially due to cryptochrome and endogenous presence of magnetic materials. Sensitivity to magnetic fields can also be involved in geomagnetic orientation by some long-distance migratory insects. In this study, near-zero magnetic fields (NZMF) in relation to normal geomagnetic fields (GMF) were setup using the Hypomagnetic Field Space System (HMFs) to investigate the effects of magnetic fields on the growth, development and reproduction of two species of migratory planthopper, the small brown planthopper (abbr. SBPH), Laodelphax striatellus, and the brown planthopper (abbr. BPH), Nilaparvata lugens. Exposure of both L. striatellus and N. lugens to NZMF delayed egg and nymphal developmental durations and decreased adult weight and female fecundity. The 1st鈥5th instars of SBPH and BPH showed different responses to NZMF. The 4th instar was significantly affected by NZMF, especially for BPH males, in which NZMF exposure reduced the difference in development duration between females and males. Compared with GMF, the vitellogenin transcript levels of newly molted female adults and the number of eggs per female were significantly reduced in both planthopper species, indicating a negative effect on fertility under NZMF. Our findings provided experimental evidence that NZMF negatively affected the growth and development of SBPH and BPH, with particularly strong effects on reproduction.
DOI:10.1016/j.anbehav.2016.08.024URL [本文引用: 2]
61Near-zero magnetic field (NZMF) enhanced positive phototaxis of adultSogatella furcifera.61NZMF affected flight capacity of adultS.02furciferain a sexually dimorphic way.61Antioxidative stress-related CRYs-circadian clock-AKH/AKHR pathway is proposed.61Reduced geomagnetic field may affect phototaxis and flight (migratory) behaviour.61We suggest a link between the probable mechanisms underlying magnetic field effects.
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DOI:10.1111/1744-7917.12256URLPMID:26227859 [本文引用: 2]
Cryptochromes (叫喊) 是蓝、紫外的轻光敏电阻器，知道在生理节奏的节奏并且在昆虫的轻依赖者的 magnetosensitivity 起关键作用。二新奇 cryptochrome 基因从棕色的 planthopper 被克隆，并且被给 Nlcry1 和 Nlcry2 的指明，与在 GenBank 的就职数字 KM108578 和 KM108579。互补 DNA 分别地 Nlcry1 定序， Nlcry2 在长度是 1935 bp 和 2463 bp，并且他们包含 1629 bp 和 1872 bp 的一个开的读物框架，与 62.53 kDa 和 70.60 kDa 的预言的分子的重量，编码 542 和 623 的氨基酸。象 DNA-photolyase 和 FAD-binding-7 领域那样的保存得好的主题在 Nlcry1 和 Nlcry2 被观察。种系发生的分析表明了 Nlcry1 和 Nlcry2 的蛋白质被聚类进昆虫 cryptochrome 1 并且 cryptochrome 2 分别地。量的聚合酶链反应证明在棕色的 planthopper 的头的送信人 RNA (mRNA ) 表示的每日的摆动为 Nlcry1 是温和的，并且为 Nlcry2 谦虚。在整个所有发展舞台， Nlcry1 和 Nlcry2 展出了极端变化和特殊表示侧面。Cryptochrome mRNA 表示在成年出现以后立即达到顶点然后随后减少了。最新出现的棕色的 planthopper 成年人的织物表示侧面比的在头显示出叫喊的更高的表示层次在胸或腹部，以及显著地在 macropterous 紧张的头的叫喊高级比在 brachypterous 紧张的头。一起拿，我们的学习的结果建议在棕色的 planthopper 描绘的二 cryptochrome 基因可能与发展生理学和移植被联系。
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DOI:10.1038/nature07183URLPMID:2559964 [本文引用: 1]
Although many animals use the Earth's magnetic field for orientation and navigation, the precise biophysical mechanisms underlying magnetic sensing have been elusive. One theoretical model proposes that geomagnetic fields are perceived by chemical reactions involving specialized photoreceptors. However, the specific photoreceptor involved in such magnetoreception has not been demonstrated conclusively in any animal. Here we show that the ultraviolet-A/blue-light photoreceptor cryptochrome (Cry) is necessary for light-dependent magnetosensitive responses in Drosophila melanogaster. In a binary-choice behavioural assay for magnetosensitivity, wild-type flies show significant naive and trained responses to a magnetic field under full-spectrum light (~300-700nm) but do not respond to the field when wavelengths in the Cry-sensitive, ultraviolet-A/blue-light part of the spectrum (<420nm) are blocked. Notably, Cry-deficient cryand cryflies do not show either naive or trained responses to a magnetic field under full-spectrum light. Moreover, Cry-dependent magnetosensitivity does not require a functioning circadian clock. Our work provides, to our knowledge, the first genetic evidence for a Cry-based magnetosensitive system in any animal.
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DOI:10.1038/ncomms5391URLPMID:25019586 [本文引用: 1]
Abstract Many higher animals have evolved the ability to use the Earth's magnetic field, particularly for orientation. Drosophila melanogaster also respond to electromagnetic fields (EMFs), although the reported effects are quite modest. Here we report that negative geotaxis in flies, scored as climbing, is disrupted by a static EMF, and this is mediated by cryptochrome (CRY), the blue-light circadian photoreceptor. CRYs may sense EMFs via formation of radical pairs of electrons requiring photoactivation of flavin adenine dinucleotide (FAD) bound near a triad of Trp residues, but mutation of the terminal Trp in the triad maintains EMF responsiveness in climbing. In contrast, deletion of the CRY C terminus disrupts EMF responses, indicating that it plays an important signalling role. CRY expression in a subset of clock neurons, or the photoreceptors, or the antennae, is sufficient to mediate negative geotaxis and EMF sensitivity. Climbing therefore provides a robust and reliable phenotype for studying EMF responses in Drosophila.
DOI:10.1371/journal.pone.0132966URLPMID:4501744 [本文引用: 1]
Abstract Although there are considerable reports of magnetic field effects (MFE) on organisms, very little is known so far about the MFE-related signal transduction pathways. Here we establish a manipulative near-zero magnetic field (NZMF) to investigate the potential signal transduction pathways involved in MFE. We show that exposure of migratory white-backed planthopper, Sogatella furcifera, to the NZMF results in delayed egg and nymphal development, increased frequency of brachypterous females, and reduced longevity of macropterous female adults. To understand the changes in gene expression underlying these phenotypes, we examined the temporal patterns of gene expression of (i) CRY1 and CRY2 as putative magnetosensors, (ii) JHAMT, FAMeT and JHEH in the juvenile hormone pathway, (iii) CYP307A1 in the ecdysone pathway, and (iv) reproduction-related Vitellogenin (Vg). The significantly altered gene expression of CRY1 and CRY2 under the NZMF suggest their developmental stage-specific patterns and potential upstream location in magnetic response. Gene expression patterns of JHAMT, JHEH and CYP307A1 were consistent with the NZMF-triggered delay in nymphal development, higher proportion of brachypterous female adults, and the shortened longevity of macropterous female adults, which show feasible links between hormone signal transduction and phenotypic MFE. By conducting manipulative NZMF experiments, our study suggests an important role of the geomagnetic field (GMF) in modulating development and physiology of insects, provides new insights into the complexity of MFE-magnetosensitivity interactions, and represents an initial but crucial step forward in understanding the molecular basis of cryptochromes and hormone signal transduction involved in MFE.
DOI:10.1016/j.cub.2005.11.030URLPMID:16332522 [本文引用: 1]
Animal flavoproteins called cryptochromes (CRYs) are generally believed to have distinct circadian clock functions in insects and mammals. We have discovered that the monarch butterfly, Danaus plexippus, has two cry genes: one encodes a fly-like protein with photosensitive properties, while the other encodes a mouse-like protein with potent transcriptional repressive activity. Database searches show that other non-drosophilid insects also have two cry genes. These findings change our view of how some insect clocks may work and redefine the evolution of animal CRYs.
DOI:10.1016/S0083-6729(08)60934-6URL [本文引用: 1]
DOI:10.1016/S0965-1748(00)00036-9URLPMID:1087610810876108 [本文引用: 1]
In the 1950s, Berta Scharrer predicted that neurosecretions from the brain regulated corpus allatum activity based upon the observation of the change in localization of neurosecretory material in the brain and change in gland activity after severance of nerves between the brain and corpus allatum. Isolation and characterization of neuropeptide regulators of juvenile hormone production by the corpora allata in the late 1980s has confirmed this prediction. Both a stimulatory allatotropin and an inhibitory allatostatin have been isolated from moth brains. Two families of allatostatins, both quite different from each other and that of moths, have been isolated from cockroaches and crickets. The wide distribution of these peptides in the nervous system, in nerves to visceral muscle, in endocrine cells of the midgut and in blood cells, indicate multifunctions in the insects in which they are allatoregulatory. Some of these other functions have been demonstrated in these insects and in insects in which these neuropeptides occur but do not act as corpus allatum regulators. For the latter group, the neuropeptide regulators of the corpora allata have yet to be isolated. The families of neurosecretory regulators will continue to grow.
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DOI:10.1016/j.tem.2004.11.003URLPMID:15620543 [本文引用: 1]
Two hormones, 20-hydroxyecdysone (20E) and juvenile hormone (JH), coordinately orchestrate insect growth and development. 20E initiates all major developmental transitions from egg, to larva, to pupa, to adult, but it is an interaction with the JH signal that transduces 20E pulses into stage-specific responses. Years of research have given us an understanding of 20E signaling pathway. By contrast, the molecular mechanism of JH action remains an enigma. Recent studies provide insight into the molecular background to JH–20E regulatory interplay. Two transcription factors – BR-C and E75A – contribute to the cross-talk between the two hormones. It appears that BR-C is a key target of JH status quo action, and E75A is a part of the mechanism whereby JH prevents BR-C activation.
DOI:10.1111/j.1365-2583.2008.00832.xURLPMID:18828836 [本文引用: 1]
After a slow start, the comparative analysis of clock genes in insects has developed into a mature area of study in recent years. Brain transplant or surgical interventions in larger insects defined much of the early work in this area, before the cloning of clock genes became possible. We discuss the evolution of clock genes, their key sequence differences, and their likely modes of regulation in several different insect orders. We also present their expression patterns in the brain, focusing particularly on Diptera, Lepidoptera, and Orthoptera, the most common non-genetic model insects studied. We also highlight the adaptive involvement of clock molecules in other complex phenotypes which require biological timing, such as social behaviour, diapause and migration.
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DOI:10.1074/jbc.M909502199URLPMID:10748136 [本文引用: 1]
In , mitochondria execute a central task in the assembly of cellular (Fe/S) proteins. The organelles synthesize their own set of Fe/S proteins, and they initiate the generation of extramitochondrial Fe/S proteins. In the present study, we identify the mitochondrial Isa1p of as a new member of the Fe/S cluster machinery. Isa1p belongs to a family of homologous proteins present in and . Deletion of the gene results in the loss of mitochondrial DNA precluding the use of the Deltaisa1 strain for functional analysis. Cells in which Isa1p was depleted by regulated gene expression maintained the mitochondrial DNA, yet the cells displayed retarded growth on nonfermentable carbon sources. This finding indicates the importance of Isa1p for mitochondrial function. Deficiency of Isa1p caused a defect in mitochondrial Fe/S protein assembly. Moreover, Isa1p was required for maturation of cytosolic Fe/S proteins. Two in a conserved sequence motif characterizing the Isa1p protein family were found to be essential for Isa1p function in the biogenesis of both intra- and extramitochondrial Fe/S proteins. Our findings suggest a function for Isa1p in the of iron or an intermediate of Fe/S cluster assembly.
DOI:10.1016/S0014-5793(00)01711-7URLPMID:10913600 [本文引用: 1]
The assembly of iron ulfur (Fe/S) clusters in a living cell is mediated by a complex machinery which, in eukaryotes, is localised within mitochondria. Here, we report on a new component of this machinery, the protein Isa2p of the yeast Saccharomyces cerevisiae. The protein shares sequence similarity with yeast Isa1p and the bacterial IscA proteins which recently have been shown to perform a function in Fe/S cluster biosynthesis. Like the Isa1p homologue, Isa2p is localised in the mitochondrial matrix as a soluble protein. Deletion of the ISA2 gene results in the loss of mitochondrial DNA and a strong growth defect. Simultaneous deletion of the ISA1 gene does not further exacerbate this growth phenotype suggesting that the Isa proteins perform a non-essential function. When Isa2p was depleted by regulated gene expression, mtDNA was maintained, but cells grew slowly on non-fermentable carbon sources. The maturation of both mitochondrial and cytosolic Fe/S proteins was strongly impaired in the absence of Isa2p. Thus, Isa2p is a new member of the Fe/S cluster biosynthesis machinery of the mitochondrial matrix and may be involved in the binding of an intermediate of Fe/S cluster assembly.
DOI:10.1016/j.cmet.2009.06.012URLPMID:19656490 [本文引用: 1]
Heme biosynthesis consists of a series of eight enzymatic reactions that originate in mitochondria and continue in the cytosol before returning to mitochondria. Although these core enzymes are well studied, additional mitochondrial transporters and regulatory factors are predicted to be required. To discover such unknown components, we utilized a large-scale computational screen to identify mitochondrial proteins whose transcripts consistently coexpress with the core machinery of heme biosynthesis. We identified SLC25A39, SLC22A4, and TMEM14C, which are putative mitochondrial transporters, as well as C1orf69 and ISCA1, which are iron-sulfur cluster proteins. Targeted knockdowns of all five genes in zebrafish resulted in profound anemia without impacting erythroid lineage specification. Moreover, silencing of Slc25a39 in murine erythroleukemia cells impaired iron incorporation into protoporphyrin IX, and vertebrate Slc25a39 complemented an iron homeostasis defect in the orthologous yeast mtm1 deletion mutant. Our results advance the molecular understanding of heme biosynthesis and offer promising candidate genes for inherited anemias.
DOI:10.1136/jmedgenet-2014-102592URLPMID:25539947 [本文引用: 1]
Abstract BACKGROUND: There are numerous nuclear genes that cause mitochondrial disorders and clinically and genetically heterogeneous disorders whose aetiology often remains unsolved. In this study, we aim to investigate an autosomal recessive syndrome causing leukodystrophy and neuroregression. We studied six patients from five unrelated consanguineous families. METHODS: Patients underwent full neurological, radiological, genetic, metabolic and dysmorphological examinations. Exome sequencing coupled with autozygosity mapping, Sanger sequencing, microsatellite haplotyping, standard and molecular karyotyping and whole mitochondrial DNA sequencing were used to identify the genetic cause of the syndrome. Immunohistochemistry, transmission electron microscopy, confocal microscopy, dipstick assays, quantitative PCR, reverse transcription PCR and quantitative reverse transcription PCR were performed on different tissue samples from the patients. RESULTS: We identified a homoallelic missense founder mutation in ISCA2 leading to mitochondrial depletion and reduced complex I activity as well as decreased ISCA2, ISCA1 and IBA57 expression in fibroblasts. MRI indicated similar white matter abnormalities in the patients. Histological examination of the skeletal muscle showed mild to moderate variation in myofibre size and the presence of many randomly distributed atrophic fibres. CONCLUSIONS: Our data demonstrate that ISCA2 deficiency leads to a hereditary mitochondrial neurodegenerative white matter disease in infancy. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.
DOI:10.1128/MCB.01963-07URLPMID:911106 [本文引用: 1]
Abstract A genome-wide screen for Saccharomyces cerevisiae iron-sulfur (Fe/S) cluster assembly mutants identified the gene IBA57. The encoded protein Iba57p is located in the mitochondrial matrix and is essential for mitochondrial DNA maintenance. The growth phenotypes of an iba57Delta mutant and extensive functional studies in vivo and in vitro indicate a specific role for Iba57p in the maturation of mitochondrial aconitase-type and radical SAM Fe/S proteins (biotin and lipoic acid synthases). Maturation of other Fe/S proteins occurred normally in the absence of Iba57p. These observations identify Iba57p as a novel dedicated maturation factor with specificity for a subset of Fe/S proteins. The Iba57p primary sequence is distinct from any known Fe/S assembly factor but is similar to certain tetrahydrofolate-binding enzymes, adding a surprising new function to this protein family. Iba57p physically interacts with the mitochondrial ISC assembly components Isa1p and Isa2p. Since all three proteins are conserved in eukaryotes and bacteria, the specificity of the Iba57/Isa complex may represent a biosynthetic concept that is universally used in nature. In keeping with this idea, the human IBA57 homolog C1orf69 complements the iba57Delta growth defects, demonstrating its conserved function throughout the eukaryotic kingdom.
