Name: measuring spatial and temporal ca2+ signals in arabidopsis plants
Uploaded: 2014-09-02T14:00:00
Description: Watch this Scientific Journal Video about Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants at JoVE.com

We are grateful to B. Stevenson for technical assistance and Dr Marc R. Knight for providing Aequorin transgenic line. This work was funded by the National Institutes of Health Grant R01 GM059138.

1，基于水母发光蛋白钙成像使用FAS系统<ol><li>准备苗木发光成像。消毒拟南芥植物表达水母发光蛋白含0.01％的Triton-100 10％的漂白粉溶液的种子。播下含有全强度MS（Murashige和Skoog基础盐混合物），1％蔗糖和1.2％琼脂的无菌种子在方形板（10×10厘米的正方形培养皿格）。代替板垂直地在分层后培养室中于4℃下进行2天（ 图1A）。 </li><li>转移到苗木电影。在7-10天龄幼苗的生长?...

<ol>
	<li>Kudla, J., Batistic, O., Hashimoto, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+signals:+the+lead+currency+of+plant+information+processing.">Calcium signals: the lead currency of plant information processing.</a> Plant Cell. 22 (3), 541-563 (2010).</li><li>Pan, Z., Zhao, Y., Zheng, Y., Liu, J., Jiang, X., Guo, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+high-throughput+method+for+screening+Arabidopsis+mutants+with+disordered+abiotic+stress-induced+calcium+signal.">A high-throughput method for screening Arabidopsis mutants with disordered abiotic stress-induced calcium signal.</a> J. Genet. Genomics. 39 (5), 225-235 (2012).</li><li>Ranf, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Defense-related+calcium+signaling+mutants+uncovered+via+a+quantitative+high-throughput+screen+in+Arabidopsis+thaliana.">Defense-related calcium signaling mutants uncovered via a quantitative high-throughput screen in Arabidopsis thaliana.</a> Mol. Plant. 5 (1), 115-130 (2012).</li><li>Zhu, X., Feng, Y., Liang, G., Liu, N., Zhu, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aequorin-based+luminescence+imaging+reveals+stimulus-+and+tissue-specific+Ca2++dynamics+in+Arabidopsis+plants.">Aequorin-based luminescence imaging reveals stimulus- and tissue-specific Ca2+ dynamics in Arabidopsis plants.</a> Mol. Plant. 6 (2), 444-455 (2013).</li><li>Knight, M. R., Campbell, A. K., Smith, S. M., Trewavas, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+plant+aequorin+reports+the+effects+of+touch+and+cold-shock+and+elicitors+on+cytoplasmic+calcium.">Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium.</a> Nature. 352 (6335), 524-526 (1991).</li><li>Van der Luit, A. H., Olivari, C., Haley, A., Knight, M. R., Trewavas, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+calcium+signalling+pathways+regulate+calmodulin+gene+expression+in+tobacco.">Distinct calcium signalling pathways regulate calmodulin gene expression in tobacco.</a> Plant Physiol. 121 (3), 705-714 (1999).</li><li>Johnson, C. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Circadian+oscillations+of+cytosolic+and+chloroplastic+free+calcium+in+plants.">Circadian oscillations of cytosolic and chloroplastic free calcium in plants.</a> Science. 269 (5232), 489-503 (1995).</li><li>Knight, H., Trewavas, A. J., Knight, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cold+calcium+signaling+in+Arabidopsis+involves+two+cellular+pools+and+a+change+in+calcium+signature+after+acclimation.">Cold calcium signaling in Arabidopsis involves two cellular pools and a change in calcium signature after acclimation.</a> Plant Cell. 8 (3), 489-503 (1996).</li><li>Logan, D. C., Knight, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mitochondrial+and+cytosolic+calcium+dynamics+are+differentially+regulated+in+plants.">Mitochondrial and cytosolic calcium dynamics are differentially regulated in plants.</a> Plant Physiol. 133 (1), 21-24 (2003).</li><li>Mehlmer, N., Parvin, N., Hurst, C. H., Knight, M. R., Teige, M., Vothknecht, U. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+toolset+of+aequorin+expression+vectors+for+in+planta+studies+of+subcellular+calcium+concentrations+in+Arabidopsis+thaliana.">A toolset of aequorin expression vectors for in planta studies of subcellular calcium concentrations in Arabidopsis thaliana.</a> J. Exp. Bot. 63 (4), 1751-1761 (2012).</li><li>Kiegle, E., Moore, C. A., Haseloff, J., Tester, M. A., Knight, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-type-specific+calcium+responses+to+drought,+salt+and+cold+in+the+Arabidopsis+root.">Cell-type-specific calcium responses to drought, salt and cold in the Arabidopsis root.</a> Plant J. 23 (2), 267-278 (2000).</li><li>Palmer, A. E., Tsien, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+calcium+signaling+using+genetically+targetable+fluorescent+indicators.">Measuring calcium signaling using genetically targetable fluorescent indicators.</a> Nat. Protoc. 1 (3), 1057-1065 (2006).</li><li>Souslova, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+fluorescent+protein-based+Ca2++sensors+with+increased+dynamic+range.">Single fluorescent protein-based Ca2+ sensors with increased dynamic range.</a> BMC. Biotechnol. 7, 37-46 (2007).</li><li>Tanaka, K., Swanson, S., Gilroy, S., Stacey, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+nucleotides+elicit+cytosolic+free+calcium+oscillations+in+Arabidopsis.">Extracellular nucleotides elicit cytosolic free calcium oscillations in Arabidopsis.</a> Plant Physiol. 154 (2), 705-719 (2010).</li><li>Iwano, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fine-tuning+of+the+cytoplasmic+Ca2++concentration+is+essential+for+pollen+tube+growth.">Fine-tuning of the cytoplasmic Ca2+ concentration is essential for pollen tube growth.</a> Plant Physiol. 150 (3), 1322-1334 (2009).</li><li>Sieberer, B., Chabaud, M., Timmers, A., Monin, A., Fournier, J., Barker, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+nuclear-targeted+Cameleon+demonstrates+intranuclear+Ca2++spiking+in+Medicago+truncatula+root+hairs+in+response+to+rhizobial+nodulation+factors.">A nuclear-targeted Cameleon demonstrates intranuclear Ca2+ spiking in Medicago truncatula root hairs in response to rhizobial nodulation factors.</a> Plant Physiol. 151 (3), 1197-1206 (2009).</li><li>Loro, G., Drago, I., Pozzan, T., Lo Schiavo, F., Zottini, M., Costa, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+of+Cameleons+to+different+subcellular+compartments+reveals+a+strict+cytoplasmic/mitochondrial+Ca2++handling+relationship+in+plant+cells.">Targeting of Cameleons to different subcellular compartments reveals a strict cytoplasmic/mitochondrial Ca2+ handling relationship in plant cells.</a> Plant J. 71 (1), 1-13 (2012).</li><li>Krebs, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=FRET-based+genetically+encoded+sensors+allow+high-resolution+live+cell+imaging+of+Ca2++dynamics.">FRET-based genetically encoded sensors allow high-resolution live cell imaging of Ca2+ dynamics.</a> Plant J. 69 (1), 181-192 (2012).</li><li>Bonza, M. C., Loro, G., Behera, S., Wong, A., Kudla, J., Costa, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analyses+of+Ca2++accumulation+and+dynamics+in+the+endoplasmic+reticulum+of+Arabidopsis+root+cells+using+a+genetically+encoded+Cameleon+sensor.">Analyses of Ca2+ accumulation and dynamics in the endoplasmic reticulum of Arabidopsis root cells using a genetically encoded Cameleon sensor.</a> Plant Physiol. 163 (3), 1230-1241 (2013).</li><li>Costa, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=H2O2+in+plant+peroxisomes:+An+in+vivo+analysis+uncovers+a+Ca2++dependent+scavenging+system.">H2O2 in plant peroxisomes: An in vivo analysis uncovers a Ca2+ dependent scavenging system.</a> Plant J. 62 (5), 760-772 (2010).</li><li>Zhu, X., Caplan, J., Mamillapalli, P., Czymmek, K., Dinesh-Kumar, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+of+endoplasmic+reticulum+calcium+ATPase+in+innate+immunity-mediated+programmed+cell+death.">Function of endoplasmic reticulum calcium ATPase in innate immunity-mediated programmed cell death.</a> EMBO J. 29 (5), 1007-1018 (2010).</li><li>Roszik, J., Lisboa, D., Sz&#246;llosi, J., Vereb, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+intensity-based+ratiometric+FRET+in+image+cytometry--approaches+and+a+software+solution.">Evaluation of intensity-based ratiometric FRET in image cytometry--approaches and a software solution.</a> Cytometry A. 75 (9), 761-767 (2009).</li><li>Kotlikoff, M. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetically+encoded+Ca2++indicators:+using+genetics+and+molecular+design+to+understand+complex+physiology.">Genetically encoded Ca2+ indicators: using genetics and molecular design to understand complex physiology.</a> J. Physiol. 578 (Pt 1), 55-67 (2007).</li><li>Tian, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+neural+activity+in+worms,+flies+and+mice+with+improved+GCaMP+calcium+indicators.">Imaging neural activity in worms, flies and mice with improved GCaMP calcium indicators.</a> Nat methods. 6 (12), 875-881 (2009).</li><li>Akerboom, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+a+GCaMP+calcium+indicator+for+neural+activity+imaging.">Optimization of a GCaMP calcium indicator for neural activity imaging.</a> J Neurosci. 32 (40), 13819-13840 (2012).</li></ol>

我们已经展示了FAS系统记录拟南芥幼苗的时空钙离子的响应。这FAS 钙记录系统提供了一个可能被适用于测量通过钙除了在此处提出的非生物胁迫的刺激各种刺激引起离子动力简便，灵敏，强健的方法。使用这个系统，我们可以很容易地在整株水平比较组织或刺激特定的时空钙离子动态。 FAS的高灵敏度允许使用低强度刺激的用于检查的 Ca 2 +的反应?...

植物细胞响应该坐标细胞动作经由信令环境。早期的细胞在应对环境刺激信号事件是一个短暂的钙增加。的图案，或一过性升高签名中游离的 Ca 2 +浓度的特征在于，其振幅，频率和持续时间。不同的时空钙签名规范不同的细胞活动1。特定刺激，如热，冷，盐，干旱，光，或植物激素，可以微调膜本地化的 Ca 2 +通道和/或转运的时空活性，导致具体的 Ca 2 +的签名。虽然钙离子转运都得到很好的特点，鲜为人知的是， 钙离子通道在植物1分子的身份和功能。对于突变体遗传筛选具有改变的 Ca 2 +响应强调刺激可能是一种有效APPRoach用于识别构成的 Ca 2 +的签名的组件。最近基于几个水母发光蛋白的 Ca 2 +检测系统已被开发，以促进遗传筛选的钙响应于病原体攻击和非生物胁迫2-4信令组件。 水母发光蛋白最早是用于检测钙在植物体内离子信号在90年代初5。从那时起，水母发光蛋白已经被定位到不同的细胞区室，如细 ​​胞质5，核6，叶绿体7，液泡膜8，线粒体9和基质10，以及不同的细胞类型中的根来监视特定的Ca 2细胞+信号11。基于水母发光蛋白钙离子的测量揭示细胞应力刺激人口的空间和时间的 Ca 2 +的响应。然而，在大多数情况下，单个细胞的钙离子的反应是unsynchroni捷思在响应组织4。因此，水母发光蛋白的 Ca 2 +的记录不一定报告在个体细胞中的钙信号。近年来，基因编码荧光蛋白（FP）为基础的 Ca 2 +的指标，如黄色卡默莱昂（YCS）12和CASEs12 13已被用于研究的 Ca 2 +具有高亚细胞分辨率的信号。 YCS是荧光共振能量转移（FRET）为基础的 Ca 2 +的指标，含有CFP和YFP的变体由钙挂钩2 +结合蛋白钙调蛋白和钙调蛋白结合肽M13。钙调蛋白发生构象变化，因为它结合的 Ca 2 +，由此带来的CFP和YFP更加靠近，从而增加了能量转移（FRET增强）。随着时间的荧光共振能量转移的水平，大约为YFP对CFP信号强度的比值计算，反映了细胞内钙离子动态。几个YC版本已被用于在植物中。 YC3.6为吨argeted到细胞质14,15，核16，线粒体17，和质膜18，和YC4.6和D4ER被定位到ER 15,19，和D3cpv被靶向过氧化物酶体20。转基因植物表达YCS允许不同类型的细胞不同的细胞内钙离子动态的活细胞成像。例（大概钙 lcium 本身 nsor）是单圆置换荧光蛋白（cpFPs）窝藏钙调蛋白和钙调蛋白结合肽M13。结合后的 Ca 2 +，情况发生构象变化，从而导致增加的荧光强度。该案例的荧光响应和Ca 2 +浓度之间的相关性使得细胞内钙离子动力学来进行定量测定。在钙 -饱和脂肪形式Case12变种12倍的增加荧光。N。 benthiminana植物瞬时EXPRessing Case12或拟南芥稳定表达病例12为材料，研究钙信号在国防和非生物胁迫4,21。异步时空钙离子振荡，细胞应对病原体的攻击，或脱水的压力已经显露了Case12基于钙离子成像。 在这里，我们提出的组织-和刺激基于水母发光蛋白发光成像特定的CA在拟南芥幼苗离子动力，并为细胞内的共焦成像与核钙的详细说明，在拟南芥根细胞离子的动态表达的FAS案例12发光成像可适于分析在完整植物或此处未描述的组织应力诱导的 Ca 2 +的动态，或筛选诱变的拟南芥属植物种群，对具有改变的应力诱导的 Ca 2 +信号的突变体。活细胞的 Ca 2 +成像设置可适于分析钙使用其他钙指标内的不同亚细胞区室或在不同类型的细胞离子动力。

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			<td height="21" style="height:21px;width:356px;">Name of Material/ Equipment</td>
			<td style="width:172px;">Company</td>
			<td style="width:175px;">Catalog Number</td>
			<td style="width:184px;">Comments/Description</td>
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			<td height="21" style="height:21px;">10X10 cm square petri dish&#160;</td>
			<td>VWR</td>
			<td style="width:175px;">60872-310</td>
			<td></td>
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			<td height="21" style="height:21px;">adhesive film&#160;</td>
			<td style="width:172px;">VWR</td>
			<td>60941-062</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;">polyethylene tubing</td>
			<td>PerkinElmer</td>
			<td style="width:175px;">9908265</td>
			<td></td>
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			<td height="21" style="height:21px;">1 mL syringe&#160;</td>
			<td style="width:172px;">VWR</td>
			<td style="width:175px;">53548-000</td>
			<td></td>
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			<td height="21" style="height:21px;">silicone grease&#160;</td>
			<td style="width:172px;">Beckman</td>
			<td style="width:175px;">335148</td>
			<td></td>
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			<td height="21" style="height:21px;">2-well chambered cover glass&#160;</td>
			<td>Nalge Nunc international</td>
			<td>155379</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:356px;">8-well chambered cover glass</td>
			<td>Nalge Nunc international</td>
			<td>155409</td>
			<td></td>
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			<td height="21" style="height:21px;">luminescence imaging system&#160;</td>
			<td>Princeton Instruments</td>
			<td style="width:175px;">N/A</td>
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			<td height="21" style="height:21px;">inverted confocal laser-scanning microscope&#160;</td>
			<td>Nikon Instruments Inc.</td>
			<td style="width:175px;">N/A</td>
			<td style="width:184px;">Nikon A1R&#160;</td>
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			<td height="21" style="height:21px;width:356px;">Imaging software</td>
			<td>Nikon Instruments Inc.</td>
			<td style="width:175px;">N/A</td>
			<td>Nikon Elements&#160;</td>
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			<td height="21" style="height:21px;width:356px;">ImageJ</td>
			<td style="width:172px;">NIH</td>
			<td style="width:175px;">N/A</td>
			<td>Public domain, Java-based image processing program</td>
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			<td height="21" style="height:21px;">DataGraph</td>
			<td>Visual Data Tools Inc</td>
			<td style="width:175px;">N/A</td>
			<td>DataGraph 3.1.1 is the newest version</td>
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		<tr height="21">
			<td height="21" style="height:21px;">coelenterazine</td>
			<td>NanoLight Technolgies</td>
			<td style="width:175px;">#301B NF-BCTZ-FB</td>
			<td></td>
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			<td height="21" style="height:21px;width:356px;">All purpose Bleach</td>
			<td style="width:172px;">Any local store</td>
			<td style="width:175px;">N/A</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:356px;">Triton X-100</td>
			<td style="width:172px;">Fisher</td>
			<td style="width:175px;">BP151500</td>
			<td></td>
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			<td height="21" style="height:21px;width:356px;">MS salt</td>
			<td style="width:172px;">Phytotechnology Labs</td>
			<td style="width:175px;">M524-50L</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:356px;">sucrose</td>
			<td style="width:172px;">VWR</td>
			<td style="width:175px;">BDH8029-12KG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:356px;">Agar</td>
			<td style="width:172px;">Sigma</td>
			<td style="width:175px;">A1296-5KG</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:356px;">Phytagel</td>
			<td style="width:172px;">Sigma</td>
			<td style="width:175px;">P8169-1KG</td>
			<td></td>
		</tr>
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</table>

measuring spatial and temporal ca2+ signals in arabidopsis plants

发展与环境线索诱发的 Ca 2 +的波动在植物细胞中。刺激特定的空间-时间的 Ca 2 +的图案是由发起的 Ca 2 +信号传导级联的细胞的 Ca 2 +结合蛋白的检测。但是，我们仍然知之甚少如何刺激产生特定的钙信号。 à 钙离子信号的特殊性可能是由于钙离子通道和/或转运活动的复杂调控响应给定的刺激。识别这些细胞成分和理解它们的功能，关键的是要使用的系统，允许的 Ca 2 +信号的灵敏和鲁棒记录同时在组织和细胞水平。基因编码的定位到不同的细胞钙指标都规定了细胞钙信号活细胞共聚焦成像的平台。在这里，我们描述的指令S为使用两钙离子检测系统：基于水母发光蛋白FAS（薄膜粘合苗）发光钙成像和case12基于活细胞共聚焦荧光钙离子成像。使用FAS系统发光成像提供了一种简单，可靠和灵敏的检测的空间和时间的 Ca 2 +信号，在组织水平的，同时采用Case12活细胞的共聚焦成像提供同时检测胞质和核的 Ca 2 +信号中的较高的分辨率。

甘露糖醇，氯化钠和H 2 O 2被用作代理进行脱水，盐和氧化应激刺激，分别。检查，如果重金属离子铜离子协同作用与任何这三个应力刺激，我们比较了的 Ca 2 +响应于在铜离子的存在或不存在各刺激。如示于图2中 ，FAS发光成像显示，Arabidoposis苗反应不同脱水，盐和氧化应激。为刺激我们研究在本研究中的浓度，75mM氯化钠诱导的第40秒的曝光（...

Watch this Scientific Journal Video about Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants at JoVE.com

Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants

测量时空的Ca<sup&gt; 2 +</sup&gt;在拟南芥信号

的 Ca 2 +信号调控植物中不同的生物过程。在这里，我们提出的方法用于监测非生物胁迫诱导的时空钙在拟南芥的细胞，并使用基因编码的钙指标水母发光蛋白或Case12组织2 +信号。

Developmental and environmental cues induce Ca2+ fluctuations in plant cells. Stimulus-specific spatial-temporal Ca2+ patterns are sensed by cellular Ca2+ ...

Research

JoVE Journal

Biology

Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants

Modeling Biological Membranes with Circuit Boards and Measuring Electrical Signals in Axons: Student Laboratory Exercises

建模生物膜与电路板，并在轴突测量电气信号：学生实验室练习

This is a demonstration of how electrical models can be used to characterize biological membranes. This exercise also introduces biophysical terminology ...

科研

生物学

Ice-Cap: A Method for Growing Arabidopsis and Tomato Plants in 96-well Plates for High-Throughput Genotyping

Ice-Cap: A Method for Growing Arabidopsis and Tomato Plants in 96-well Plates for High-Throughput Genotyping

冰帽：为发展方法拟南芥和番茄植株在96孔板高通量的基因分型

It is becoming common for plant scientists to develop projects that require the genotyping of large numbers of plants. The first step in any genotyping ...

Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging

Measuring Intracellular Ca2+ Changes in Human Sperm using Four Techniques: Conventional Fluorometry, Stopped Flow Fluorometry, Flow Cytometry and Single Cell Imaging

测量细胞内Ca<sup&gt; +</sup在人类精子的变化采用4项技术：传统的荧光，停流荧光，流式细胞术及单细胞成像

Spermatozoa  are male reproductive cells especially designed to reach, recognize and fuse  with the egg. To perform these tasks, sperm cells must be ...

Automated Analysis of Dynamic Ca2+ Signals in Image Sequences

Automated Analysis of Dynamic Ca2+ Signals in Image Sequences

动态钙的自动分析<sup&gt; 2 +</sup&gt;在图像序列信号

Intracellular Ca2+ signals are commonly studied with fluorescent Ca2+ indicator dyes and microscopy techniques. However, quantitative analysis of Ca2+ ...

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

Imaging Local Ca2+ Signals in Cultured Mammalian Cells

成像本地CA<sup&gt; 2+</sup&gt;在培养的哺乳动物细胞信号

Cytosolic Ca2+ ions regulate numerous aspects of cellular activity in almost all cell types, controlling processes as wide-ranging as gene transcription, ...

Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor

Real-time In Vivo Recording of Arabidopsis Calcium Signals During Insect Feeding Using a Fluorescent Biosensor

实时在体记录了拟南芥钙信号在使用荧光传感器的昆虫饲喂时

Calcium ions are predicted to be key signaling entities during biotic interactions, with calcium signaling forming an established part of the plant ...

Applications of Spatio-temporal Mapping and Particle Analysis Techniques to Quantify Intracellular Ca2+ Signaling In Situ

Applications of Spatio-temporal Mapping and Particle Analysis Techniques to Quantify Intracellular Ca2+ Signaling In Situ

时空映射和粒子分析技术在细胞内 ca2 +原位信令定量中的应用

Ca2+ imaging of isolated cells or specific types of cells within intact tissues often reveals complex patterns of Ca2+ signaling. This activity requires ...

Laser-Capture Microdissection RNA-Sequencing for Spatial and Temporal Tissue-Specific Gene Expression Analysis in Plants

激光捕获微分块RNA-测序空间和时间组织特异性基因表达分析

The development of a complex multicellular organism is governed by distinct cell types that have different transcriptional profiles. To identify ...

Wide-Field, Real-Time Imaging of Local and Systemic Wound Signals in Arabidopsis

Wide-Field, Real-Time Imaging of Local and Systemic Wound Signals in Arabidopsis

阿拉伯多普西斯局部和系统伤口信号的广域实时成像

Plants respond to mechanical stresses such as wounding and herbivory by inducing defense responses both in the damaged and in the distal undamaged parts. ...

An Easy and Flexible Inoculation Method for Accurately Assessing Powdery Mildew-Infection Phenotypes of Arabidopsis and Other Plants

一种简单灵活的接种方法，用于准确评估拟南芥和其他植物的白粉病感染表型

Reducing crop losses due to fungal diseases requires improved understanding of the mechanisms governing plant immunity and fungal pathogenesis, which in ...