Name: quantitative measurement of intrathecally synthesized proteins in mice
Uploaded: 2019-11-29T13:00:00
Description: Watch this Scientific Journal Video about Quantitative Measurement of Intrathecally Synthesized Proteins in Mice at JoVE.com

作者感谢达特茅斯的比较医学和研究中心（CCMR）的工作人员对用于这些研究的小鼠进行专家护理。伯恩斯坦研究基金资助了这项研究。

所有动物工作都采用达特茅斯盖塞尔医学院机构动物护理和使用委员会 （IACUC） 审查和批准的协议。
1. 流体收集
注：血清和CSF均是必需的。生存和尸体解剖需要每个流体收集两个协议。
<ol><li>使用生存程序收集血清和CSF 注：对于生存液体收集，血清收集应先于CSF收集，因为它是一个侵入性较低的程序。CSF 必须在血清抽取后一周内获得。<ol><li>血清?...

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评估CSF蛋白质浓度增加的定量方法是中枢神经系统生理和病理状态表征的有用工具。然而，除了可靠的CSF蛋白质水平定量外，CSF蛋白的检测还需要表达在CSF中区分血液和CNS衍生成分的结果。然而，迄今为止，常用的蛋白质定量测定不允许区分两种蛋白质成分，即使借助高通量工具也是如此。因此，为了区分在CNS隔间中合成的蛋白质和从血液中提取的蛋白质，提出了对蛋白质测量的具体校正。这种?...

脑脊液（CSF）是一种围绕大脑和脊髓的透明无色液体，具有重要的临床和基本科学意义。CSF保持中枢神经系统（CNS）的电解环境，平衡全身酸碱状态，为神经元和胶质细胞提供营养，作为中枢神经系统的淋巴系统发挥作用，并在CNS1中传输激素、神经递质、细胞因子和其他神经肽。因此，由于CSF成分反映了中枢神经系统的活性，这种流体提供了一个有价值的，但间接的，以表征的中枢神经系统的生理和病理状态。
CSF已经用于诊断影响中枢神经系统的疾病超过一百年，在大部分时间，它主要是由临床医生作为诊断工具研究。然而，近年来，神经生物学家已经认识到CSF在研究中枢神经系统病理生理学方面的潜力。特别是，神经科学领域引入了几种高通量蛋白分析工具，以便对CSF的蛋白质组成进行详细研究，并期望这种分析有助于深入了解动态变化发生在CNS内。
Luminex和Simoa技术2，3等多倍免疫测定技术的技术发展，使今天的研究人员能够以极低的浓度检测数百种蛋白质。此外，这些相同的技术允许使用小样本量，从而促进对小动物，包括小鼠的研究，在小动物中，CSF的有限样本量直到最近才排除了对液体的详细特征。
然而，一个警告是，在CSF中测量的蛋白质可能来自细胞内合成和/或从血清转出由于血脑界面（BBI）受损。不幸的是，仅CSF的蛋白质分析只能确定这两个成分的总和。为了区分转体蛋白和内蛋白，必须根据血清浓度的个体变异性以及屏障完整性调整使用任何可用蛋白质分析工具的CSF蛋白测量。然而，虽然这种调整在临床实践中是常用的，例如CSF IgG指数，该指数对检测细胞内IgG合成4、5、6具有高灵敏度，但迄今为止很少有研究能够纠正CSF蛋白浓度对血清浓度和屏障完整性的7、8。
目前，Reibergram方法是确定蛋白质屏障功能和宫内合成的最佳方法。它是CSF/血清商图中的图形化评估，以综合的方式分析屏障（dys）功能和宫内蛋白合成，指纯血衍生蛋白质9，10。高度丰富的蛋白质白蛋白通常被选择作为参考蛋白，因为它只在肝脏中产生，因为它的大小，大约70kDa，是小蛋白和大蛋白质11之间的中间体。分析图最初由赖伯和费尔根豪尔于1987年对免疫球蛋白（Igs）11的主要类别进行定义，根据对数千个人类样本9的分析结果进行实证分析。该方法随后被两个菲克扩散定律应用于分子扩散/流速理论12得到证实。这种理论证明蛋白质通过屏障扩散具有双曲分布，可以定量解释中枢神经系统9、13中蛋白质的动态。总体而言，使用 Reibergram 来演示片内蛋白合成的优点是，它同时识别从血清进入 CSF 的蛋白质分数，以及由于本地生产而在 CSF 中发现的蛋白质量。
本文和相关协议描述了从CSF和血液采集到最终计算校正CSF蛋白水平，用于神经小鼠小鼠模型中的细胞内蛋白合成的定量测量。障碍。此过程提供了一个基准，用于评估 （1） 任何 CSF 蛋白质的病理生理学来源和 （2） 屏障完整性的稳定性和功能意义。这个程序和协议不仅可用于评估小鼠CSF样本，而且可用于分析多种动物模型的神经系统疾病和人类患者的CSF。

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		<tr height="21">
			<td height="21" style="height:21px;width:327px;">1 mL insulin syringe</td>
			<td style="width:184px;">BD</td>
			<td style="width:127px;">329650</td>
			<td style="width:847px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1 mL syringe</td>
			<td style="width:184px;">BD</td>
			<td style="width:127px;">329622</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">25 gauge needle</td>
			<td style="width:184px;">BD</td>
			<td style="width:127px;">305122</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">3 mL syringe</td>
			<td style="width:184px;">BD</td>
			<td style="width:127px;">309582</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">30 gauge insulin needle</td>
			<td style="width:184px;">BD</td>
			<td style="width:127px;">305106</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Absorbent pads</td>
			<td style="width:184px;">Any suitable brand</td>
			<td style="width:127px;"></td>
			<td></td>
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			<td height="21" style="height:21px;width:327px;">Acepromazine</td>
			<td style="width:184px;">Patterson Vet Supply Inc</td>
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			<td height="21" style="height:21px;width:327px;">BioPlex Handheld Magnetic Washer</td>
			<td style="width:184px;">BioRad</td>
			<td style="width:127px;">171020100</td>
			<td>Magnet</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">BioPlex MAGPIX Multiplex Reader</td>
			<td style="width:184px;">BioRad</td>
			<td style="width:127px;">171015001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">BioPlex Pro Flat Bottom Plates</td>
			<td style="width:184px;">BioRad</td>
			<td style="width:127px;">171025001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Biotinilated detection antibody</td>
			<td style="width:184px;">Any suitable source</td>
			<td style="width:127px;"></td>
			<td>The antibody has to be directed against the species of the protein of interest.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Bovine Serum Albumin (BSA)</td>
			<td style="width:184px;">Sigma</td>
			<td>A4503</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Buprenorphine hydrochloride</td>
			<td style="width:184px;">PAR Pharmaceutical</td>
			<td style="width:127px;">NDC 42023-179-05</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Capillary Tubes</td>
			<td style="width:184px;">Sutter Instrument</td>
			<td style="width:127px;">B100-75-10</td>
			<td>OD: 1.0 mm, ID: 0.75 mm Borosilicate glass 10 cm; drawn over Bunsen to make ID smaller.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Centrifuge tube, 0.2 mL</td>
			<td style="width:184px;">VWR</td>
			<td style="width:127px;">20170-012</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Centrifuge tube, 0.5 mL</td>
			<td style="width:184px;">VWR</td>
			<td style="width:127px;">87003-290</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Centrifuge tube, 1.5 mL</td>
			<td style="width:184px;">VWR</td>
			<td style="width:127px;">87003-294</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Chlorhexidine diacetate</td>
			<td style="width:184px;">Nolvasan</td>
			<td style="width:127px;">E004272</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Disposable pipettes tips</td>
			<td style="width:184px;">Any suitable brand</td>
			<td style="width:127px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Ear bars</td>
			<td style="width:184px;">KOPF Instruments</td>
			<td style="width:127px;">1921 or 1922</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Ethanol</td>
			<td style="width:184px;">Kopter</td>
			<td style="width:127px;">V1001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Freezer</td>
			<td style="width:184px;">VWR</td>
			<td style="width:127px;">VWR32086A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Gauze</td>
			<td style="width:184px;">Medline</td>
			<td style="width:127px;">NON25212</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Heating pad</td>
			<td style="width:184px;">Sunbeam</td>
			<td style="width:127px;"></td>
			<td>XL King Size SoftTouch, 4 Heat Settings with Auto-Off, Teal, 12-Inch x 24-Inch</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Induction Chamber</td>
			<td style="width:184px;">VETEQUIP</td>
			<td style="width:127px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Isoflurane</td>
			<td style="width:184px;">Patterson Vet Supply Inc</td>
			<td style="width:127px;">NDC 14043-704-06</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Ketamine (KetaVed)</td>
			<td style="width:184px;">Patterson Vet Supply Inc</td>
			<td style="width:127px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MagPlex Microspheres (antibody-coupled)</td>
			<td style="width:184px;">BioRad</td>
			<td style="width:127px;"></td>
			<td style="width:847px;">Antibody-coupled magnetic bead</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Microplate Shaker</td>
			<td style="width:184px;">Southwest Scientific</td>
			<td>SBT1500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Microretractors</td>
			<td style="width:184px;">Carfill Quality</td>
			<td style="width:127px;">ACD-010</td>
			<td>Blunt - 1 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Microsoft Office (Excel)</td>
			<td style="width:184px;">Microsoft</td>
			<td style="width:127px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">MilliPlex MAP Mouse Immunoglobulin Isotyping Magnetic Bead Panel</td>
			<td style="width:184px;">EMD Millipore</td>
			<td style="width:127px;">MGAMMAG-300K</td>
			<td>Commercial kit for the quantification through Luminex of a panel of immunoglobulin isotypes and subclasses in mouse fluids.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Mouse Albumin capture ELISA kit</td>
			<td style="width:184px;">Novus Biological</td>
			<td style="width:127px;">NBP2-60484</td>
			<td>Commercial kit for the quantification through ELISA of albumin in mouse fluids.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Multichannel pipette</td>
			<td style="width:184px;">Eppendorf</td>
			<td>3125000060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Non-Sterile swabs</td>
			<td style="width:184px;">MediChoice</td>
			<td style="width:127px;">WOD1002</td>
			<td>Need to be autoclaved for sterility</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Oxygen</td>
			<td style="width:184px;">AIRGAS</td>
			<td style="width:127px;">OX USPEA</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Pasteur Pippettes</td>
			<td style="width:184px;">Fisher</td>
			<td style="width:127px;">13-678-20A</td>
			<td>5 &#38; 3/4&#34;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">PDS suture with disposable needle, 6-0 Prolen</td>
			<td style="width:184px;">Patterson Vet</td>
			<td style="width:127px;">8695G</td>
			<td>P-3 Reverse Cutting, 18&#34;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">PE-Streptavidin</td>
			<td style="width:184px;">BD Biosciences</td>
			<td style="width:127px;">554061</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Pipetters</td>
			<td style="width:184px;">Eppendorf</td>
			<td style="width:127px;">Research seriers</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polyethylene tubing</td>
			<td style="width:184px;"></td>
			<td style="width:127px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Refrigerated Centrifuge</td>
			<td style="width:184px;">Beckman Coulter</td>
			<td style="width:127px;">ALLEGRA X-12R</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Scale</td>
			<td style="width:184px;">Uline</td>
			<td style="width:127px;">H2716</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Scalpel</td>
			<td style="width:184px;">Feather</td>
			<td style="width:127px;">EF7281</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Shaver</td>
			<td style="width:184px;">Harvard Apparatus</td>
			<td style="width:127px;">52-5204</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Standard proteins</td>
			<td>Any suitable source</td>
			<td></td>
			<td>The best choice for a reference standard is a purified, known concentration of the protein of interest.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Stereotaxic instrument</td>
			<td style="width:184px;">KOPF Instruments</td>
			<td style="width:127px;">Model 900LS</td>
			<td>Standard Accessories</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Sterile 1 x PBS</td>
			<td style="width:184px;">Corning Cellgro</td>
			<td style="width:127px;">21-040-CV</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Sterile saline</td>
			<td style="width:184px;">Baxter</td>
			<td style="width:127px;">0338-0048-02</td>
			<td>0.9 % Sodium Chloride Irrigation USP</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Surgical Forceps Curved, 7 (2)</td>
			<td style="width:184px;">Fine Science Tools</td>
			<td style="width:127px;">11271-30</td>
			<td>Dumont</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Surgical Scissors</td>
			<td style="width:184px;">Fine Science Tools</td>
			<td style="width:127px;">14094-11</td>
			<td>Stainless 25x</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Vaporizer + Flow meter</td>
			<td style="width:184px;">Moduflex Anhestesia Instruments</td>
			<td style="width:127px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Vortex</td>
			<td style="width:184px;">Fisher</td>
			<td>02-215-414</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Warming pad</td>
			<td style="width:184px;">Kent Scientific Corporation</td>
			<td style="width:127px;">RT-JR-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Water Sonicator</td>
			<td style="width:184px;">Cole Parmer</td>
			<td>EW-08895-01</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Xylazine</td>
			<td style="width:184px;">Patterson Vet Supply Inc</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

quantitative measurement of intrathecally synthesized proteins in mice

脑脊液（CSF）是一种存在于大脑和脊髓中的液体，对基础科学和临床科学都非常重要。CSF蛋白质成分的分析为基础神经科学研究以及神经系统疾病提供了关键信息。需要注意的一点是，在CSF中测量的蛋白质可能同时来自血清的术中合成和转位，而CSF的蛋白质分析只能确定这两个组分的总和。为了区分动物模型和人类血液内产生的蛋白质转位和蛋白质，CSF蛋白质分析测量使用传统的蛋白质分析工具必须包括白蛋白CSF/血清商（Q白蛋白）的计算，这是血脑界面（BBI）完整性的标志，以及蛋白质指数（Q蛋白/Q白蛋白），这是钙蛋白内合成的估计。该协议说明了从CSF和血液采集到商数和指数计算的整个过程，用于定量测量神经疾病小鼠模型中的细胞内蛋白质合成和BBI损伤。

这项具有代表性的实验旨在比较多发性硬化症（MS）两个临床相关啮齿动物模型中IgG的宫内合成：PLP139-151-诱导的复发性自免疫性脑脊髓炎（R-EAE）和慢性渐进性、泰勒的小鼠脑脊髓炎病毒诱发脱骨髓性疾病（TMEV-IDD）。R-EAE是理解复发性MS的有用模型，而TMEV-IDD模型具有慢性渐进性MS19。
在本研究中，对R-EAE（n = 12）和TMEV-IDD（n = 28）中的宫内IgG合成进?...

Watch this Scientific Journal Video about Quantitative Measurement of Intrathecally Synthesized Proteins in Mice at JoVE.com

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

脊髓液蛋白水平升高可能是血浆蛋白在改变的血脑屏障上扩散的结果，也可以是血脑内合成的结果。本文提出了一种优化的测试方案，有助于区分这两种情况，并提供对内代谢蛋白的定量测量。

小鼠体内合成蛋白的定量测量

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

该协议允许CSF和血液采集在CSF蛋白质水平的定量校正中测量成神经紊乱的小鼠模型中的蛋白质合成。该程序提供了一个基线，据此可以评估感兴趣的CSF蛋白质的病理生理来源，以及血液CSF蛋白完整性的稳定性和功能意义。间间蛋白质合成和屏障完整性分析可应用于其他动物模型和人类研究。例如，检查中枢神经系统的疾病。在小鼠中收集大量清洁CSF在技术上具有挑战性，因此建议在获得大量未污染样品之前练习该技术。演示这些程序的将是来自达特茅斯实验和分子医学课程的研究生Mymeal Linzey，我们新的免疫学研究小组。用于通过逆行轨道出血收集血清。在确认超过15克麻醉小鼠对踏板反射缺乏反应后，用拇指和食指抓住耳朵后面的松弛皮肤，用无显性手的食指，并使用食指拉回眼睛上方的皮肤。用拇指将皮肤拉回眼睛下方，将巴斯德移液器的尖端（以大约 45 度角）放入眼球下方的眼窝中，朝向眼窝中间，同时在手指之间旋转移液器。然后施加短暂、温和的压力并释放，让血液进入移液器。采集血液样本后，轻轻切除毛细管，不伤害眼睛，将血液转移到1.5毫升离心管。关闭眼睑后，用纱布施加轻度压力，以防止进一步出血。让血液在室温下凝固30至60分钟，然后通过离心使样品旋转下来。使用清洁移液器技术，将分离的血清收集到标有 500 微升的新小瓶中，并立即将血清冷冻在 80 摄氏度。在确认对踏板反射缺乏反应后，去除足够大的头发区域，以便立即在麻醉小鼠头骨的骨端采集脑脊髓液。在无菌条件下将鼠标放在立体技术装置的易发位置，用耳条稳定头部。用30%氯己丁二甲酸酯的手术部位，使下垂的皮肤切口低于奥奇普特，以暴露肌肉覆盖西斯特纳磁石。使用钳子，钝解剖皮下组织和肌肉，并使用微缩回器保持肌肉分开，以暴露杜拉母体脑膜层在西斯特纳磁石。用无菌 PBS 轻轻清洗组织，以清除任何可能的血液污染，并使用无菌棉签将杜拉母体擦干。将初始穿刺定位在 cisterna 磁石中对于获得丰富的、未受污染的 CSF 至关重要。使用 30 量针，轻轻刺穿覆盖 cisterna 磁石的膜，并快速轻轻地将小玻璃毛细管插入穿刺中。收集5至12微升的CSF时，小心地将管从膜上取下，并使用一块聚乙烯管将管子连接到三毫升注射器。将收集的CSF注射到冰上标有500微升的管中，并使用一次性针头和各种多二酮缝合线关闭切口。清洁任何干血或组织的区域，将鼠标放在干净、温暖的笼中，并进行监测，直到完全保持。通过离心收集 CSF，目视检查颗粒并超浸其血液污染的任何迹象。然后使用清洁移液器技术，将含CSF的超盐转移到新的200微升管中，然后用PBS以一到三比稀释CSF，在80摄氏度时立即储存。对于通过心脏穿刺采集血清，在CSF采集后立即将鼠标放在超供位置。用70%的酒精擦拭腹部皮肤。使用剪刀打开胸腔，露出心脏，并将连接到 3 毫升注射器的 25 针插入左心室。然后轻轻地对注射器柱塞施加负压，在采集血液后取走针头。压压柱塞，将采集的血液喷射到1.5毫升小瓶中。现在，让血液在室温下凝固30至60分钟，然后通过离心分离血清。然后，使用清洁移液器技术，将血清转移到新的标签 500 微升小瓶中，在 80 摄氏度下立即储存。要量化匹配血清和CSF样本中的目标蛋白质和白蛋白，请使用标准蛋白质定量测定，使用参考标准蛋白质为感兴趣的每种蛋白质准备标准曲线。分析后，将原始数据导出到相应的软件图形程序，并绘制检测信号荧光强度与标准蛋白质浓度，为感兴趣的每种蛋白质创建标准曲线。然后，使用标准曲线计算样本中每个感兴趣的分析物的浓度。最后，使用白蛋白和目标分析物浓度计算Q值和内部指数。与相应的年龄匹配假对对相比，两个经测试多发性硬化症模型的CSF中，总IgG的实际水平显著增加。R-EAE小鼠表现出显著增强的白细胞商值，表明这些小鼠的血脑屏障的渗透性有所提高。相反，TMEV-IDD和假老鼠之间在白细胞商之间没有差异，这证实了以前关于TMEV-IDD小鼠存在完整屏障的发现。此外，在TMEV-IDD动物中，IgG指数值显著上升，因此，观察到IGG内部生产。蛋白质指数的计算有助于识别对神经炎病和神经退行性疾病的早期诊断、结果预测和疾病过程监测有用的新型蛋白质生物标志物。

Research

JoVE Journal

Neuroscience

The Vermicelli and Capellini Handling Tests: Simple quantitative measures of dexterous forepaw function in rats and mice

粉条和Capellini处理测试：在大鼠和小鼠的灵巧的前爪功能简单的定量措施

Previous characterizations of rodent eating behavior have revealed that they use coordinated forepaw movements to manipulate food pieces. We have extended ...

科研

神经科学

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

Quantitative Measurement of Relative Retinoic Acid Levels in E8.5 Embryos and Neurosphere Cultures Using the F9 RARE-Lacz Cell-based Reporter Assay

使用F9稀土LACZ基于Cell的报告基因检测在E8.5胚胎和神经球文化相对维甲酸水平的定量测量

Retinoic acid (RA) is an important developmental morphogen that coordinates anteroposterior and dorsoventral axis patterning, somitic differentiation, ...

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

In Vivo Gene Transfer to Schwann Cells in the Rodent Sciatic Nerve by Electroporation

体内基因转移到雪旺细胞在啮齿动物坐骨神经电穿孔

The formation of the myelin sheath by Schwann cells (SCs) is essential for rapid conduction of nerve impulses along axons in the peripheral nervous ...

Quantitative Measurement of &#947;-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms

基于细胞荧光检测平台的γ-分泌介导的淀粉样前体蛋白和缺口裂解的定量测定

We have developed a pair of cell-based reporter gene assays to quantitatively measure &#947;-secretase cleavage of distinct substrates. This manuscript ...

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila

嗜德维人神经元树突状树突状复杂度的定量分析

Dendrites are the branched projections of a neuron, and dendritic morphology reflects synaptic organization during the development of the nervous system. ...

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

Quantitative Cell Biology of Neurodegeneration in Drosophila Through Unbiased Analysis of Fluorescently Tagged Proteins Using ImageJ

荧光标记蛋白 ImageJ 的无偏分析在果蝇变性定量细胞生物学中的应用

With the rising prevalence of neurodegenerative diseases, it is increasingly important to understand the underlying pathophysiology that leads to neuronal ...

Imaging and Analysis of Neurofilament Transport in Excised Mouse Tibial Nerve

切除小鼠神经神经纤维神经丝的成像与分析

Neurofilament protein polymers move along axons in the slow component of axonal transport at average speeds of ~0.35-3.5 mm/day. Until recently the study ...

Electrocorticographic Recording of Cerebral Cortex Areas Manipulated Using an Adeno-Associated Virus Targeting Cofilin in Mice

使用腺相关病毒在小鼠中瞄准科菲林操纵的大脑皮层区域的电皮质记录

The use of electrocorticographic (ECoG) recordings in rodents is relevant to sleep research and to the study of a wide range of neurological conditions. ...

Semi-Quantitative Determination of Dopaminergic Neuron Density in the Substantia Nigra of Rodent Models using Automated Image Analysis

使用自动图像分析对啮齿动物模型的亚斯坦蒂娅尼格拉多巴明神经元密度进行半定量测定

Estimation of the number of dopaminergic neurons in the substantia nigra is a key method in pre-clinical Parkinson's disease research. Currently, unbiased ...