基于鼠精细胞的味觉阻抗传感器用于苦味物质检测的研究
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摘要
本文设计了一种基于雄鼠精细胞的细胞阻抗传感器用于苦味物质的特异性检测。雄鼠精细胞内含有大量T2Rs受体(G蛋白偶联受体)可以对苦味物质产生特异性响应,苦味受体被苦味物质激活后产生的响应会引起细胞形态骨架的变化,从而可以被细胞阻抗传感器检测。本文探索了实验的最佳细胞密度,检测了苯硫脲和奎宁两种常见的苦味物质的响应,苯硫脲检测范围为10μmol/L~200μmol/L,检出限为4μmol/L;奎宁检测范围为62.5μmol/L~1 000μmol/L,检出限为40μmol/L。传感器阻抗值增量与苦味物质浓度呈一定的线性相关性。此外,论文对该味觉传感器的特异性进行了测试分析,表明了基于细胞传感器的检测方法为代替动物和人的苦味测试提供了一种可能的新的手段。
In this study,the feasibility of utilizing male mouse germ cells in testis and electrical cell-substrate impedance sensor(ECIS)was explored to build a cell-based bitter biosensor,which can sensitively and specifically respond to bitter compounds. Male mouse germ cells express lots of bitter receptor T2Rs(G protein coupled receptors)which can sensitively and specifically respond to bitter compounds,when active by ligands will affect cell morphology in a specific manner. The best cell density was explored;cell-impedance response profiles of germ cells to two bitter compounds were investigated by analyzing the response intensity under various concentrations,The linear detection range of PTC is 10 μmol/L~200 μmol/L,the detection limit is 4 μmol/L;the linear detection range of quinine is 62.5 μmol/L~1 000 μmol/L,the detection limit is 4 μmol/L; Furthermore,impedance responses to five basic tastes were examined to evaluate the specificity of cell-based bitter biosensor in bitter detection. The results revealed that this hybrid bitter biosensor could respond to those bitter compounds in a dose-dependent manner. And it could respond to bitter compounds specifically. This biosensor may provide a promising approach for detecting various bitter compounds.
In this study,the feasibility of utilizing male mouse germ cells in testis and electrical cell-substrate impedance sensor(ECIS)was explored to build a cell-based bitter biosensor,which can sensitively and specifically respond to bitter compounds. Male mouse germ cells express lots of bitter receptor T2Rs(G protein coupled receptors)which can sensitively and specifically respond to bitter compounds,when active by ligands will affect cell morphology in a specific manner. The best cell density was explored;cell-impedance response profiles of germ cells to two bitter compounds were investigated by analyzing the response intensity under various concentrations,The linear detection range of PTC is 10 μmol/L~200 μmol/L,the detection limit is 4 μmol/L;the linear detection range of quinine is 62.5 μmol/L~1 000 μmol/L,the detection limit is 4 μmol/L; Furthermore,impedance responses to five basic tastes were examined to evaluate the specificity of cell-based bitter biosensor in bitter detection. The results revealed that this hybrid bitter biosensor could respond to those bitter compounds in a dose-dependent manner. And it could respond to bitter compounds specifically. This biosensor may provide a promising approach for detecting various bitter compounds.
引文
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[7]Xu J,Cao J,Iguchi N,et al.Functional Characterization of BitterTaste Receptors Expressed in Mammalian Testis[J].Molecular Human Reproduction,2013,19(1):17-28.
[8]Li F,Zhou M.Depletion of Bitter Taste Transduction Leads to Massive Spermatid Loss in Transgenic Mice[J].Molecular Human Reproduction,2012,18(6):289-297.
[9]Hu L,Xu J,Qin Z,et al.Detection of Bitterness in Vitro by a Novel Male Mouse Germ Cell-Based Biosensor[J].Sensors and Actuators B:Chemical,2016,223()461-469.
[10]Kammermann M,Denelavas A,Imbach A,et al.Impedance Measurement:A New Method to Detect Ligand-Biased Receptor Signaling[J].Biochemical and Biophysical Research Communications,2011,412(3):419-424.
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[12]Ghosh P M,Keese C R,Giaever I.Morphological Response of Mammalian Cells to Pulsed AC Fields[J].Bioelectrochemistry and Bioenergetics,1994,33(2):121-133.
[13]Giaever I,Keese C.Monitoring Fibroblast Behavior in Tissue Culture with an Applied Electric Field[J].Proceedings of the National Academy of Sciences,1984,81(12):3761-3164.
[14]Xiao C,Lachance B,Sunahara G,et al.An In-Depth Analysis of Electric Cell-Substrate Impedance Sensing to Study the Attachment and Spreading of Mammalian Cells[J].Analytical Chemistry,2002,74(6):1333-1339.
[15]Asphahani F,Wang K,Thein M,et al.Single-Cell Bioelectrical Impedance Platform for Monitoring Cellular Response to Drug Treatment[J].Physical Biology,2011,8(1):015006.
[16]苏凯麒,邹玲,王琴,等.基于细胞阻抗传感器的腹泻性毒素检测系统设计与实现[J].传感技术学报,2014,27(3):283-288.
[17]王天星,黎洪波,苏凯麒,等.基于细胞电阻抗传感器的细胞多生理参数分析系统设计[J].传感技术学报,2014,27(12):1589-1595.
[18]Abassi Y A,Jackson J A,Zhu J,et al.Label-Free,Real-Time Monitoring of Ig E-Mediated Mast Cell Activation on Microelectronic Cell Sensor Arrays[J].Journal of Immunological Methods,2004,292(1):195-205.
[19]Abassi Y A,Xi B,Zhang W,et al.Kinetic Cell-Based Morphological Screening:Prediction of Mechanism of Compound Action and Off-Target Effects[J].Chemistry&biology,2009,16(7):712-23.