氯化镉暴露对斑马鱼幼鱼神经行为毒性作用
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摘要
为探索低剂量镉持续暴露对斑马鱼胚胎和幼鱼神经发育的影响,将受精后36h(36hpf)的斑马鱼胚胎暴露于不同浓度氯化镉受试液中持续至受精后6d(6dpf)。在一般毒性观察的基础上,采用高通量视频跟踪技术检测了48孔板中幼鱼的自发运动能力和对外界刺激产生的惊恐反应及恢复能力。结果显示:在不引起外观形态明显改变的暴露剂量下幼鱼自发运动表现异常。其中,0.1～0.5μmol·L-1暴露组幼鱼自发运动增多、轨迹紊乱、刻板式转向运动增加、触壁活动减少;1.0μmol·L-1暴露组幼鱼出现运动抑制、静止和原地震颤等状态,幼鱼的自发运动距离与氯化镉浓度关系呈现剂量依赖性倒"U"型,这种行为毒性效应与乙醇急性暴露引起的运动效应相似。光照惊恐反射实验中,暴露组幼鱼对突发的光暗变化表现敏感或迟钝。在撤除光源后暗适应减慢或加快。以上结果说明,斑马鱼胚胎和幼鱼对氯化镉的神经发育毒性作用敏感,并且高通量的幼鱼神经行为学方法在外源性化学物的毒性评价中能够提高检测效率,具有良好的应用前景。
To realize the developmental neurotoxicity of cadmium on zebrafish embryos and larvae,the neurobehavioral response of individual zebrafish larva ( 6 days post-fertilization) in 48-well microtiter plates was investigated after cadmium chloride ( 0. 5-2. 0 μmol·L-1 ) administered to the embryos ( 36 hours post-fertilization) by immersion. Movement of larvae were recorded and analyzed by using a video-tracking system. Results showed that low-concentration cadmium chloride evokes the spontaneous movement tracks of larvae disorder. The tereotypic circling swimming of larvae increased,but the touching wall response reduced in range of 0. 1-0. 5 μmol·L-1 exposure dose. Larvae represented motorial inhibition,tremor and freezing at 1. 0 μmol·L-1 cadmium chloride. Moreover,the distance moved of larvae showed the "Inverted U" dose-response to the concentration of cadmium chloride,which the locomotor activity pattern was similar to that caused by acute ethanol exposure. Light-evoked startle escape response results showed that cadmium chloride can promote or reduce the sensibility of larvae to light or dark transition. Accordingly,the habituation of activity presented postpone or accelerate in dark after light stimulation. These results demonstrated that the development of nerve of zebrafish embryos and larvae were sensitive to toxicity of cadmium chloride. In addition,zebrafish larvae were a type of powerful model in developmental neurotoxicity of exogenous assessment by using high-throughput neurobehavioral detections.
To realize the developmental neurotoxicity of cadmium on zebrafish embryos and larvae,the neurobehavioral response of individual zebrafish larva ( 6 days post-fertilization) in 48-well microtiter plates was investigated after cadmium chloride ( 0. 5-2. 0 μmol·L-1 ) administered to the embryos ( 36 hours post-fertilization) by immersion. Movement of larvae were recorded and analyzed by using a video-tracking system. Results showed that low-concentration cadmium chloride evokes the spontaneous movement tracks of larvae disorder. The tereotypic circling swimming of larvae increased,but the touching wall response reduced in range of 0. 1-0. 5 μmol·L-1 exposure dose. Larvae represented motorial inhibition,tremor and freezing at 1. 0 μmol·L-1 cadmium chloride. Moreover,the distance moved of larvae showed the "Inverted U" dose-response to the concentration of cadmium chloride,which the locomotor activity pattern was similar to that caused by acute ethanol exposure. Light-evoked startle escape response results showed that cadmium chloride can promote or reduce the sensibility of larvae to light or dark transition. Accordingly,the habituation of activity presented postpone or accelerate in dark after light stimulation. These results demonstrated that the development of nerve of zebrafish embryos and larvae were sensitive to toxicity of cadmium chloride. In addition,zebrafish larvae were a type of powerful model in developmental neurotoxicity of exogenous assessment by using high-throughput neurobehavioral detections.
引文
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[2]梁友信.劳动卫生与职业病学[M].4版.北京:人民卫生出版社,2000:167-681
[3]Weiss B,Spyker J M.The susceptibility of the fetus and child to chemical pollutants:Behavioral implications of prenatal and early postnatal exposure to chemical pollu-tants[J].Pediatrics,1974,53(5):851-859
[4]Chisolm Jr J J.Heavy metal exposures:Toxicity from metal-metal interactions,and behavioral effects[J].Pediatrics,1974,53(5):841-843
[5]Thatcher R W,Lester M L,McAlaster R,et al.Effects of low levels of cadmium and lead on cognitive functio-ning in children[J].Archives of Environmental Health,1982,37(3):159-166
[6]Dési I,Nagymajtényi L,Schulz H.Behavioural and neurotoxicological changes caused by cadmium treatment of rats during development[J].Journal of Applied Tox-icology,1998,18(1):63-70
[7]朱俊德,余资江,戈果,等.慢性镉中毒对小鼠学习记忆及海马CA3区的影响[J].环境与健康杂志,2009,26(7):576-579Zhu J D,Yu Z J,Ge G,et al.Effects of chronic cadmi-um exposure on learning,memory and hippocampus CA3in mice[J].Journal of Environment and Health,2009,26(7):576-579(in Chinese)
[8]Baranski B.Effect of prenatal exposure to cadmium on avoidance acquisition in rats[J].Medycyna Pracy,1983,34(5-6):381-383
[9]Ali M M,Murthy R C,Chandra S V.Developmental and long term neurobehavioral toxicity of low level in-utero cadmium exposure in rats[J].Neurobehavior Toxicoloy and Teratology,1986,8(5):463-468
[10]Linney E,Upchurch L,Donerly S.Zebrafish as a neu-rotoxicological model[J].Neurotoxicology and Teratol-ogy,2004,26(6):709-718
[11]Chow E S,Hui M N,Lin C C,et al.Cadmium inhibits neurogenesis in zebrafish embryonic brain development[J].Aquatic Toxicology,2008,87(3):157-169
[12]邹苏琪,殷梧,杨昱鹏,等.斑马鱼行为学实验在神经科学中的应用[J].生物化学与生物物理进展,2009,36(1):5-12Zou S Q,Yin W,Yang Y P,et al.The ethology appli-cation of zebrafish in neuroscience[J].Progress in Bi-ochemistry and Biophysics,2009,36(1):5-12(in Chinese)
[13]Tierney K B.Behavioural assessments of neurotoxic effects and neurodegeneration in zebrafish[J].Biochimica et Biophysica Acta,2011,1812(3):381-389
[14]MacPhail R C,Brooks J,Hunter D L,et al.Locomotion in larval zebrafish:Influence of time of day,lighting and eth-anol[J].Neurotoxicology,2009,30(1):52-58
[15]Westerfield M.The Zebrafish Book(4th)[M].Eu-gene:University of Oregon Press,2000:28-196
[16]Kimmel C,Ballard W W,Kimmel S R,et al.Stages of embryonic development in the zebrafish[J].Develop-mental Dynamics,1995,203(3):253-310
[17]Parng C,Roy N M,Ton C,et al.Neurotoxicity assess-ment using zebrafish[J].Journal of Pharmacological and Toxicological Methods,2007,5(1):103-112
[18]Agency for Toxic Substances and Disease Registry(ATSDR).Toxicological Profile for Cadmium[R].Atlanta:U.S.Department of Health and Human Serv-ices,1999
[19]Cheng S H,Wai A W K,So C H,et al.Cellular and molecular basis of cadmium-induced deformities in ze-brafish embryos[J].Environmental Toxicology and Chemistry,2000,19(12):3024-3031
[20]Blechinger S R,Kusch R C,Haugo K,et al.Brief em-bryonic cadmium exposure induces a stress response and cell death in the developing olfactory system followed by long-term olfactory deficits in juvenile zebrafish[J].Toxicology and Applied Pharmacology,2007,224(1):72-80
[21]Matsui J L,Egana A L,Sponboltz T R,et al.Effects of ethanol on photoreceptors and visual function in develo-ping zebrafish[J].Investigative Opthalmology&Visual Science,2006,47(10):4589-4597
[22]Carvan III M J,Loucks E,Weber D N,et al.Ethanol effects on the developing zebrafish:Neurobehavior and skeletal morphogenesis[J].Neurotoxicology and Tera-tology,2004,26(6):757-768
[23]Sylvain N J,Brewster D L,Ali D W.Zebrafish embryos exposed to alcohol undergo abnormal development of motor[J].Neurotoxicology and Teratology,2010,32(1):472-480
[24]Irons T D,MacPhail R C,Hunter D L,et al.Acute neuroactive drug exposures alter locomotor activity in larval zebrafish[J].Neurotoxicology and Teratology,2010,32(1):84-90
[25]Easter S S,Nicola G N.The development of vision in the zebrafish(Danio rerio)[J].Developmental Biolo-gy,1996,180(2):646-663
[26]史慧勤,张利军,彭辉,等.氯丙嗪在斑马鱼胚胎和早期幼鱼发育过程中的神经毒性作用[J].中国比较医学杂志,2011,21(10-11):150-156Shi H Q,Zhang L J,Peng H,et al.Developmental neurotoxicity effect of chlorpromazine on zebrafish em-bryo and larvae[J].Chinese Journal of Comparative Medicine,2011,21(10-11):150-156(in Chinese)
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