Name: a method to study &#945;-synuclein toxicity and aggregation using a humanized yeast model
Uploaded: 2022-11-25T12:30:00
Description: Watch this Scientific Journal Video about A Method to Study Î±-Synuclein Toxicity and Aggregation Using a Humanized Yeast Model at JoVE.com

我们感谢James Bardwell和Tiago F. Outeiro分享含有α-突触核蛋白的质粒。李昌汉获得了韩国政府（MSIT）资助的韩国国家研究基金会（NRF）（赠款2021R1C1C1C1011690），教育部资助的NRF基础科学研究计划（赠款2021R1A6A1A1A10044950）和亚洲大学的新教师研究基金。

1. 培养基和溶液的制备
<ol><li>培养基制备<ol><li>要制备YPD培养基，将50gYPD粉末溶解在dH2O中，使最终体积为1L.高压灭菌器。在室温 （RT） 下储存。</li>
		<li>要制作YPD琼脂培养基，将50gYPD粉末和20g琼脂溶解在1L dH2O.高压灭菌器中。冷却后，倒入培养皿上。储存在4°C。</li>
		<li>要用棉子糖（SRd）-Ura培养基制备SC，将6.7g酵母氮碱与硫酸铵溶解在795mL的dH2O.高压灭菌器...

<ol>
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A., Cherry, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Yeast+as+a+model+organism.">Yeast as a model organism.</a> Science. 277 (5330), 1259-1260 (1997).</li><li>Kachroo, A. H., Vandeloo, M., Greco, B. M., Abdullah, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+yeast+to+model+human+biology,+disease+and+evolution.">Humanized yeast to model human biology, disease and evolution.</a> Disease Models &amp; Mechanisms. 15 (6), (2022).</li><li>Outeiro, T. 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A., Xu, H., Golding, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+mRNA+copy+number+in+individual+Escherichia+coli+cells+using+single-molecule+fluorescent+in+situ+hybridization.">Measuring mRNA copy number in individual Escherichia coli cells using single-molecule fluorescent in situ hybridization.</a> Nature Protocols. 8 (6), 1100-1113 (2013).</li><li>Tenreiro, S., Munder, M. C., Alberti, S., Outeiro, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Harnessing+the+power+of+yeast+to+unravel+the+molecular+basis+of+neurodegeneration.">Harnessing the power of yeast to unravel the molecular basis of neurodegeneration.</a> Journal of Neurochemistry. 127 (4), 438-452 (2013).</li><li>Nielsen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Yeast+systems+biology:+Model+organism+and+cell+factory.">Yeast systems biology: Model organism and cell factory.</a> Biotechnology Journal. 14 (9), 1800421 (2019).</li><li>Eleutherio, 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=Oxidative+stress+and+aging:+learning+from+yeast+lessons.">Oxidative stress and aging: learning from yeast lessons.</a> Fungal Biology. 122 (6), 514-525 (2018).</li><li>Rencus-Lazar, S., DeRowe, Y., Adsi, H., Gazit, E., Laor, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Yeast+models+for+the+study+of+amyloid-associated+disorders+and+development+of+future+therapy.">Yeast models for the study of amyloid-associated disorders and development of future therapy.</a> Frontiers in Molecular Biosciences. 6, 15 (2019).</li><li>Franco, R., Rivas-Santisteban, R., Navarro, G., Pinna, A., Reyes-Resina, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genes+implicated+in+familial+Parkinson's+disease+provide+a+dual+picture+of+nigral+dopaminergic+neurodegeneration+with+mitochondria+taking+center+stage.">Genes implicated in familial Parkinson's disease provide a dual picture of nigral dopaminergic neurodegeneration with mitochondria taking center stage.</a> International Journal of Molecular Sciences. 22 (9), 4643 (2021).</li><li>Wan, O. W., Chung, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+alpha-synuclein+oligomerization+and+aggregation+in+cellular+and+animal+models+of+Parkinson's+disease.">The role of alpha-synuclein oligomerization and aggregation in cellular and animal models of Parkinson's disease.</a> PLoS One. 7 (6), 38545 (2012).</li><li>Xu, L., Pu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alpha-synuclein+in+Parkinson's+disease:+From+pathogenetic+dysfunction+to+potential+clinical+application.">Alpha-synuclein in Parkinson's disease: From pathogenetic dysfunction to potential clinical application.</a> Parkinson's Disease. 2016, 1720621 (2016).</li><li>Gitler, A. D., 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=The+Parkinson's+disease+protein+&#945;-synuclein+disrupts+cellular+Rab+homeostasis.">The Parkinson's disease protein &#945;-synuclein disrupts cellular Rab homeostasis.</a> Proceedings of the National Academy of Sciences of the United States of America. 105 (1), 145-150 (2008).</li><li>Sampson, T. R., 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=A+gut+bacterial+amyloid+promotes+&#945;-synuclein+aggregation+and+motor+impairment+in+mice.">A gut bacterial amyloid promotes &#945;-synuclein aggregation and motor impairment in mice.</a> Elife. 9, 53111 (2020).</li><li>Evans, M. 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鉴于人类各种细胞系统的复杂性，使用酵母作为研究人类神经退行性疾病的模型是有利的。虽然使用酵母研究人脑的复杂细胞相互作用几乎是不可能的，但从单细胞的角度来看，酵母细胞在基因组序列同源性和基本的真核细胞过程方面与人类细胞具有高度的相似性8，13。此外，鉴于遗传操作的便利性，酵母可以促进复杂和苛刻的人类系统的研究。因此?...

帕金森病（PD）是全球老龄化社会的一个严重问题。α-突触核蛋白的聚集与PD密切相关，α-突触核蛋白的蛋白质聚集体被广泛用作诊断疾病的分子生物标志物1。α-突触核蛋白是一种小的酸性蛋白质（长度为 140 个氨基酸），具有三个结构域，即 N 端脂质结合α螺旋、淀粉样蛋白结合中心结构域 （NAC） 和 C 端酸性尾部2。α-突触核蛋白的错误折叠可以自发发生，并最终导致淀粉样蛋白聚集体的形成，称为路易体3。α-突触核蛋白可能以多种方式促进PD的发病机制。一般来说，人们认为其异常的可溶性低聚形式称为原纤维是有毒物种，通过影响各种细胞靶标（包括突触功能）导致神经元细胞死亡3。
用于研究神经退行性疾病的生物学模型必须在基因组和细胞生物学方面与人类相关。最好的模型是人类神经元细胞系。然而，这些细胞系与一些技术问题有关，例如培养物维护困难，转染效率低以及费用高4。由于这些原因，需要一个简单可靠的工具来加速这一研究领域的进展。重要的是，该工具应该易于用于分析收集的数据。从这些角度来看，各种模式生物已被广泛使用，包括 果蝇、 秀丽隐杆线虫、 Danio rerio、酵母和啮齿动物5。其中，酵母是最好的模式生物，因为基因操作很容易，而且比其他模式生物便宜。最重要的是，酵母与人类细胞具有高度相似性，例如与人类直系同源物有60%的序列同源性，与人类疾病相关基因6的近源性为25%，并且它们还共享基本的真核细胞生物学。酵母含有许多蛋白质，其序列和功能与人体细胞相似7。事实上，表达人类基因的酵母已被广泛用作阐明细胞过程的模型系统8。这种酵母菌株被称为人源酵母，是探索人类基因功能的有用工具9。人源化酵母在研究遗传相互作用方面具有优点，因为遗传操作在酵母中已经建立起来。
本研究以酿酒酵母为模式生物研究PD的发病机制，特别是研究α-突触核蛋白聚集体形成和细胞毒性10。为了在出芽酵母中表达α-突触核蛋白，使用W303a菌株进行质粒转化，质粒编码α-突触核蛋白的野生型和家族性PD相关变体。由于W303a菌株在URA3上具有营养突变，因此适用于选择含有URA3质粒的细胞。质粒中编码的α-突触核蛋白的表达在GAL1启动子下调节。因此，可以控制α-突触核蛋白的表达水平。此外，绿色荧光蛋白（GFP）在α-突触核蛋白C末端区域的融合可以监测α-突触核蛋白病灶的形成。为了了解α-突触核蛋白家族性PD相关变异的特征，我们还在酵母中表达这些变异并检查了它们的细胞效应。该系统是筛选具有保护作用的化合物或基因的简单工具，可防止α-突触核蛋白的细胞毒性。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>96 well plate</td>
			<td>SPL</td>
			<td>30096</td>
			<td></td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>TAESHIN</td>
			<td>0158</td>
			<td></td>
		</tr>
		<tr>
			<td>Bacto Agar</td>
			<td>BD Difco</td>
			<td>214010</td>
			<td></td>
		</tr>
		<tr>
			<td>Breathe-easy</td>
			<td>diversified biotech</td>
			<td>BEM-1</td>
			<td>Gas permeable sealing membrane for microtiter plates</td>
		</tr>
		<tr>
			<td>cover glasses</td>
			<td>Marienfeld</td>
			<td></td>
			<td>24 x 60 mm</td>
		</tr>
		<tr>
			<td>Culture tube</td>
			<td>SPL</td>
			<td>40014</td>
			<td></td>
		</tr>
		<tr>
			<td>Cuvette</td>
			<td>ratiolab</td>
			<td>2712120</td>
			<td></td>
		</tr>
		<tr>
			<td>D-(+)-Galactose</td>
			<td>sigma</td>
			<td>G0625</td>
			<td></td>
		</tr>
		<tr>
			<td>D-(+)-Glucose</td>
			<td>sigma</td>
			<td>G8270</td>
			<td></td>
		</tr>
		<tr>
			<td>D-(+)-Raffinose pentahydrate</td>
			<td>Daejung</td>
			<td>6638-4105</td>
			<td></td>
		</tr>
		<tr>
			<td>Incubator (shaking)</td>
			<td>Labtron</td>
			<td></td>
			<td>model: SHI1</td>
		</tr>
		<tr>
			<td>Incubator (static)</td>
			<td>Vision scientific</td>
			<td></td>
			<td>model: VS-1203PV-O</td>
		</tr>
		<tr>
			<td>LiAc</td>
			<td>sigma</td>
			<td>L6883</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate reader</td>
			<td>Tecan</td>
			<td>30050303 01</td>
			<td>Model: Infinite 200 pro</td>
		</tr>
		<tr>
			<td>multichannel pipette 20-200 &#181;L</td>
			<td>gilson</td>
			<td>FA10011</td>
			<td></td>
		</tr>
		<tr>
			<td>multichannel pipette 2-20 &#181;L</td>
			<td>gilson</td>
			<td>FA10009</td>
			<td></td>
		</tr>
		<tr>
			<td>Olympus microscope</td>
			<td>Olympus</td>
			<td>IX-53</td>
			<td></td>
		</tr>
		<tr>
			<td>PEG</td>
			<td>sigma</td>
			<td>P4338</td>
			<td>average mol wt 3,350</td>
		</tr>
		<tr>
			<td>Petridish</td>
			<td>SPL</td>
			<td>10090</td>
			<td></td>
		</tr>
		<tr>
			<td>pRS426</td>
			<td></td>
			<td></td>
			<td>Christianson, T. W., Sikorski, R. S., Dante, M., Shero, J. H. &#38; Hieter, P. Multifunctional yeast high-copy-number shuttle vectors. Gene. 110 (1), 119-122 (1992).</td>
		</tr>
		<tr>
			<td>pRS426 GAL1 promoter &#945;-synuclein A30P</td>
			<td></td>
			<td></td>
			<td>Outeiro, T. F. &#38; Lindquist, S. Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science. 302 (5651), 1772-1775 (2003)</td>
		</tr>
		<tr>
			<td>pRS426 GAL1 promoter &#945;-synuclein A53T</td>
			<td></td>
			<td></td>
			<td>Outeiro, T. F. &#38; Lindquist, S. Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science. 302 (5651), 1772-1775 (2003)</td>
		</tr>
		<tr>
			<td>pRS426 GAL1 promoter &#945;-synuclein E46K</td>
			<td></td>
			<td></td>
			<td>Outeiro, T. F. &#38; Lindquist, S. Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science. 302 (5651), 1772-1775 (2003)</td>
		</tr>
		<tr>
			<td>pRS426 GAL1 promoter &#945;-synuclein WT</td>
			<td></td>
			<td></td>
			<td>Outeiro, T. F. &#38; Lindquist, S. Yeast cells provide insight into alpha-synuclein biology and pathobiology. Science. 302 (5651), 1772-1775 (2003)</td>
		</tr>
		<tr>
			<td>Reservoir</td>
			<td>SPL</td>
			<td>23050</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>eppendorf</td>
			<td>6131 05560</td>
			<td></td>
		</tr>
		<tr>
			<td>W303a</td>
			<td></td>
			<td></td>
			<td>Present from James Bardwell</td>
		</tr>
		<tr>
			<td>Yeast nitrogen base w/o amino acids</td>
			<td>Difco</td>
			<td>291940</td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast synthetic drop-out medium supplements without uracil</td>
			<td>sigma</td>
			<td>Y1501</td>
			<td></td>
		</tr>
		<tr>
			<td>YPD</td>
			<td>Condalab</td>
			<td>1547.00</td>
			<td></td>
		</tr>
	</tbody>
</table>

a method to study &#945;-synuclein toxicity and aggregation using a humanized yeast model

帕金森病是第二常见的神经退行性疾病，其特征是由含有错误折叠和聚集的α突触核蛋白的路易体的形成引起的进行性细胞死亡。α-突触核蛋白是一种丰富的突触前蛋白，可调节突触囊泡运输，但其蛋白质内含物的积累会导致神经毒性。最近的研究表明，包括细菌伴侣在内的各种遗传因素可以在体外减少α-突触核蛋白聚集体的形成。然而，监测细胞中的抗聚集作用也很重要，以将其作为患者的潜在治疗方法。使用神经元细胞是理想的，但这些细胞难以处理，需要很长时间才能表现出抗聚集表型。因此，需要一种快速有效的体内工具进一步评估体内抗聚集活性。此处描述的方法用于监测和分析表达人α突触核蛋白的人源化酵母酿酒酵母中的抗聚集表型。该协议展示了可用于监测α-突触核蛋白诱导的细胞毒性以及细胞中α-突触核蛋白聚集体形成的体内工具。

已知α-突触核蛋白的高表达与PD模型系统中的神经元细胞死亡和PD有关。本研究描述了三种监测α-突触核蛋白细胞毒性和酵母中聚集α-突触核蛋白病灶形成的方法。在这里，α-突触核蛋白在酵母中过表达，并检查了野生型α突触核蛋白和三种被称为PD家族突变体的α-突触核蛋白变体的表型（图1 和 材料表）。
pRS426载体中 GAL1 启动子下的?...

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A Method to Study &#945;-Synuclein Toxicity and Aggregation Using a Humanized Yeast Model

需要α-突触核蛋白的 体内 生理模型来研究和了解帕金森病的发病机制。我们描述了一种使用人源化酵母模型监测α-突触核蛋白的细胞毒性和聚集体形成的方法。

一种基于人源酵母模型研究α-突触核蛋白毒性和聚集的方法

Parkinson's disease is the second most common neurodegenerative disorder and is characterized by progressive cell death caused by the formation of Lewy ...

这是一种监测R-pass细胞核表型和特征的简单方法。它也经常可以在全基因组研究中恢复，以找到影响R-pass细胞核稳定性的遗传因素。由于这是一种成熟的运动生物，因此与使用人类细胞系或动物模型相比，该技术很容易并且不需要太多时间和精力。R-pass细胞核的聚集与帕金森病密切相关。可能影响R-pass细胞核稳定性的蛋白质或化学物质可以在这种人源化酵母模型中进行测试。首先，将三个单菌落贴在SD-URA琼脂平板上，以选择含有α-突触核蛋白质粒的细胞。然后，将每株三个贴片的酵母转化体接种到 3 毫升 SRD-URA 中，用于用 2% 棉子糖选择性生长含有质粒的酵母，以抑制 Gal1 启动子和 0.1% 葡萄糖下 α-突触核蛋白的诱导。将接种的培养物在30摄氏度下以每分钟200转的速度搅拌孵育。第二天，将过夜培养的细胞接种到3毫升新鲜SRD-URA中，直到它们在600纳米处达到0.2的光密度。然后，将培养物在30摄氏度下以每分钟200转的速度搅拌孵育6小时。使用分光光度计测量600纳米处的光密度。接下来，在96孔板的泳道1中用无菌蒸馏水将样品稀释至600纳米处的光密度为0.5，以平衡每个培养物中的细胞数。通过向接下来的五个泳道中加入 160 微升无菌蒸馏水，对酵母细胞进行 5 倍连续稀释，并确保在每个泳道中连续稀释细胞时总体积为 40 微升。使用多通道移液器从每个孔转移10微升，以在SD-URA琼脂平板上发现样品以进行对照，并使用SG-URA琼脂平板监测毒性。将斑点板在 30 摄氏度下倒置孵育四天。每24小时拍摄一次平板图像，直到第四天，然后通过比较肉眼发现的菌株之间的生长差异来分析数据。将每个菌株的三个修补的酵母转化体接种到三毫升 SRD-URA 中，并在 30 摄氏度下以每分钟 200 转的速度孵育过夜。第二天，测量在600纳米下培养过夜的细胞的光密度，并将细胞接种到96孔板中总共200微升新鲜SD-URA和SG-URA中。将包括细胞在内的最终体积调整为200微升，并将初始细胞光密度在600纳米处调整为0.05。调整微量滴定板中的程序设置。将目标温度设置为30摄氏度，间隔时间为15分钟，振荡持续时间设置为890秒，振荡幅度设置为5毫米，测量为600纳米处的吸光度。将板孵育2-3天，以使所有菌株在酶标仪中到达固定相。通过绘制增长曲线来分析数据。将修补的酵母转化体接种到 3 毫升 SRD-URA 中，并在 30 摄氏度下以每分钟 200 转的速度搅拌培养过夜。第二天，将过夜培养的细胞重新接种到3毫升新鲜SRD-URA中，在600纳米处的光密度为0.003，然后在30摄氏度下以每分钟200转的速度搅拌孵育培养物，直到光密度达到0.2。然后加入300微升20%半乳糖，然后在30摄氏度下以每分钟200转的速度搅拌下再孵育6小时。通过以800g离心5分钟收集一毫升细胞培养物，并弃去上清液。将细胞沉淀轻轻重悬于30微升蒸馏水中。在载玻片上准备琼脂糖凝胶垫。重悬细胞并在载玻片上加载五微升细胞重悬，并使用切割的琼脂糖垫覆盖细胞进行检查。要使用100倍物镜进行显微成像，请使用屏幕捕获选项使用该程序生成图像，并选择明场以使用浸油使用100倍物镜聚焦细胞。切换到GFP荧光滤光片，并将图像曝光时间调整为50毫秒至300毫秒之间，通过平衡信噪比获得清晰的图像。打开图像文件，通过使用图像分析软件根据细胞中的病灶数量对细胞进行计数来分析数据。PRS426载体中Gal1启动子下的突触核蛋白表达在含有半乳糖作为诱导剂的琼脂平板上表现出明显的生长迟缓。在液体培养物中也观察到突触核蛋白表达的生长差异。在两种培养条件下，野生型突触核蛋白和具有E46K或A53T突变的变体由于突触核蛋白毒性而显示出生长缺陷。然而，不形成聚集体的突触核蛋白A30p变体显示出无毒表型。表现出严重细胞毒性的菌株显示出突触核蛋白聚集体的病灶，但在A30p变体中，毒性较小的突触核蛋白形式扩散到整个细胞中。为了获得可重复的结果，重要的是在每个实验中使用处于相同生长阶段的细胞，尤其是在监测突触核蛋白相关表型时。正如我们在荧光显微镜数据中所显示的，突触核蛋白可以形成更大的聚集体。因此，它可以简单地通过离心分离，也可以通过使用突触核蛋白特异性抗体的蛋白质印迹进行定量。该技术适用于高通量筛选，以发现影响突触核蛋白聚集的新遗传因子和化合物。因此，我们希望找到新的帕金森病治疗候选药物。

Research

JoVE Journal

Biology

Genetic Studies of Human DNA Repair Proteins Using Yeast as a Model System 

人类DNA修复蛋白的遗传研究利用酵母作为模型系统

Understanding the roles of human DNA repair proteins in genetic pathways is a formidable challenge to many researchers.  Genetic studies in mammalian ...

科研

生物学

Platelet Adhesion and Aggregation Under Flow using Microfluidic Flow Cells

根据流量的血小板粘附和聚集，使用微流控流细胞

Platelet aggregation occurs in response to vascular injury where the extracellular matrix below the endothelium has been exposed. The platelet adhesion ...

Hi-C: A Method to Study the Three-dimensional Architecture of Genomes.

嗨 - C：一个方法研究基因组的三维架构。

The three-dimensional folding of chromosomes compartmentalizes the genome and and can bring distant functional elements, such as promoters and enhancers, ...

Growth Assays to Assess Polyglutamine Toxicity in Yeast

生长实验，以评估在酵母中的多聚谷氨酰胺毒性

Protein misfolding is associated with many human diseases, particularly neurodegenerative diseases, such as Alzheimer&#x2019;s disease, Parkinson's ...

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

Using Caenorhabditis elegans as a Model System to Study Protein Homeostasis in a Multicellular Organism

The folding and assembly of proteins is essential for protein function, the long-term health of the cell, and longevity of the organism. Historically, the ...

Generating a &#34;Humanized&#34; Drosophila S2 Cell Line Sensitive to Pharmacological Inhibition of Kinesin-5

Generating a "Humanized" Drosophila S2 Cell Line Sensitive to Pharmacological Inhibition of Kinesin-5

生成一个“人性化”<em&gt;果蝇</em&gt; S2细胞敏感的驱动蛋白-5的药理抑制

Kinetochores are large protein-based structures that assemble on centromeres during cell division and link chromosomes to spindle microtubules. Proper ...

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

Using a Whole-mount Immunohistochemical Method to Study the Innervation of the Biliary Tract in Suncus murinus

使用全身免疫组织化学方法研究胆道的神经支配<em&gt; Suncus murinus</em

This work describes the whole-mount immunohistochemistry staining method in detail, using neurofilament protein antibody to label the innervation of the ...

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans

Methods to Study Changes in Inherent Protein Aggregation with Age in Caenorhabditis elegans

方法研究线虫线虫中固有的蛋白质聚集与年龄的变化

In the last decades, the prevalence of neurodegenerative disorders, such as Alzheimer's disease (AD) and Parkinson's disease (PD), has grown. These ...

Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes

新型杀虫化学对蚊子毒性测试规程

New classes of insecticides with novel modes of action are needed to control insecticide resistant populations of mosquitoes that transmit diseases such ...