大鼠背根神经元中Ca~(2+)的激光共聚焦显微成像与定量分析Imaging and Quantitative Analysis of Ca~(2+) in Dorsal Root Ganglion Neurons by Confocal Laser Scanning Microscopy
沈秀球;黄义梅;杨洪钦;郑莉琴;王瑜华;谢树森;
摘要(Abstract):
成功实现体外培养背根神经元(Dorsal Root Ganglion,DRG),利用差速贴壁法进行提纯,通过细胞免疫荧光技术鉴定所培养的细胞纯度,发现其纯度高达90%,完全适合运用于细胞分子机理研究.在此基础上,采用激光共聚焦显微成像实验观察DRG内的Ca2+荧光信号,并对Ca2+浓度进行了定量分析.研究结果有助于深入探究和定量分析神经元中Ca2+参与特定的细胞生理功能.
关键词(KeyWords): 背根神经节(DRG);神经元;疼痛;Ca2+;fluo-3/AM
基金项目(Foundation): 国家自然科学基金资助项目(60978071);; 福建省自然科学基金资助项目(2010J01322)
作者(Authors): 沈秀球;黄义梅;杨洪钦;郑莉琴;王瑜华;谢树森;
参考文献(References):
- [1]Babes A,Zorzon D,Reid G.Two populations of cold-sensitive neurons in rat dorsal root ganglia and their modulationby nerve growth factor[J].European Journal of Neuroscience,2004,20(9):2276-2282.
- [2]Shu X,Mendell L M.Nerve growth factor acutely sensitizes the response of adult rat sensory neurons to capsaicin[J].Neuroscience Letters,1999,274(3):159-162.
- [3]Farinas I,Cano-Jaimez M,Bellmunt E,et al.Regulation of neurogenesis by neurotrophins in developing spinalsensory ganglia[J].Brain Research Bulletin,2002,57(6):809-816.
- [4]Mamet J,Baron A,Lazdunski M,et al.Proinflammatory mediators stimulators of sensory neuron excitability viathe expression of acid-sensing ion channels[J].The Journal of Neuroscience,2002,22(24):10662-10670.
- [5]Sacha A M,Derek C M,Richard K,et al.Glial cell line-derived neurotrophic factor family members sensitizenociceptors in vitro and produce thermal hyperalgesia in vivo[J].The Journal of Neuroscience,2006,26(33):8588-8599.
- [6]Taherzadeh O,Otto W R,Anand U,et al.Influence of human skin injury on regeneration of sensory neurons[J].Cell and Tissue Research,2003,312(3):275-280.
- [7]Malin S A,Davis B M,Molliver D C.Production of dissociated sensory neuron cultures and considerations for theiruse in studying neuronal function and plasticity[J].Nature Protocols,2007,2(1):152-160.
- [8]Anirvan G,Michael E G.Calcium signaling in neurons:molecular mechanisms and cellular consequences context[J].Science,1995,268(5208):239-247.
- [9]陈蓓蕾,于布为.Ca2+钙离子在疼痛和抗伤害性感受中的作用[J].国际麻醉学与复苏杂志,2006,27(3):170-173.
- [10]Baccaglini P I,Hogan P G.Some rat sensory neurons in culture express characteristics of differentiated painsensory cells[J].Proceedings of the National Academy of Sciences USA,1983,80(2):594-598.
- [11]Schuichi Koizumi,Kayoko Fujishita,Kaori Inoue,et al.Ca2+waves in keratinocytes are transmitted to sensoryneurons:the involvement of extracellular ATP and P2Y2 receptor activation[J].Biochemical Journal,2004,380(2):329-338.
- [12]Liu Xiaohong,Zeng Junwei,Zhao Yandong,et al.Inhibition of ATP-induced Ca2+influx by corticosterone indorsal root ganglion neurons[J].Neurochemical Research,2010,35(5):804-810.
- [13]王丽琴,宋学琴,王晓娟,等.大鼠背根神经节细胞的纯化培养[J].脑与神经疾病杂志,2003,11(5):265-267.
- [14]隋峰,杜新亮,张畅斌,等.一种原代培养大鼠DRG神经元的新方法[J].中国药理学通报,2009,25(7):971-973.
- [15]McDonough S I,Cseresnyes Z,Schneider M F.Origin sites of calcium release and calcium oscillations in frogsympathetic neurons[J].The Journal of Neuroscience,2000,20(24):9059-9070.
- [16]张小玲,鲍霞,贾宏瑛,等.细胞Ca2+测定研究进展[J].咸阳师范学院学报,2002,17(6):19-21.
- [17]Barry D B,Doouglas L,Jeffery D K.Intranuclear Ca2+transients during neurite regeneration of an adultmammalian neuron[J].Neurobiology,1992,89(17):7978-7982.