Name: assessment of blood-brain barrier permeability by intravenous infusion of fitc-labeled albumin in a mouse model of neurodegenerative disease
Uploaded: 2017-11-08T14:00:00
Description: Watch this Scientific Journal Video about Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease at JoVE.com

这项工作得到了 "基金会 Neuromed" 的支持, 由意大利卫生部 "74年2月 Corrente" 资助科诺诺夫

所有动物程序均由 IRCCS Neuromed 动物保育审查委员会和 #34 批准; 依 Superiore Sanit 和 #224; #34; (许可证编号: 1163/2015-PR), 并按照欧盟指令2010的指导方针/63/欧洲动物实验. 
 1. 注射到颈静脉内的 FITC-白蛋白溶液的制备 注意: 所有实验都是在清单 (11 周龄) HD R6/2 小鼠和年龄和性别匹配的野生型 (重量) 控制中进行的。窝. 
<ol> <li> 在磷酸缓冲盐水 (PBS) 中溶解 FITC-?...

<ol>
	<li>Obermeier, B., Verma, A., Ransohoff, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+blood-brain+barrier.">The blood-brain barrier.</a> Handb Clin Neurol. 133, 39-59 (2016).</li><li>Serlin, Y., Shelef, I., Knyazer, B., Friedman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomy+and+physiology+of+the+blood-brain+barrier.">Anatomy and physiology of the blood-brain barrier.</a> Semin Cell Dev Biol. 38, 2-6 (2015).</li><li>Moretti, 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=Blood-brain+barrier+dysfunction+in+disorders+of+the+developing+brain.">Blood-brain barrier dysfunction in disorders of the developing brain.</a> Front Neurosci. 9, 40 (2015).</li><li>Zhao, Z., Nelson, A. R., Betsholtz, C., Zlokovic, B. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishment+and+Dysfunction+of+the+Blood-Brain+Barrier.">Establishment and Dysfunction of the Blood-Brain Barrier.</a> Cell. 163 (5), 1064-1078 (2015).</li><li>Drouin-Ouellet, 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=Cerebrovascular+and+blood-brain+barrier+impairments+in+Huntington's+disease:+Potential+implications+for+its+pathophysiology.">Cerebrovascular and blood-brain barrier impairments in Huntington's disease: Potential implications for its pathophysiology.</a> Ann Neurol. 78 (2), 160-177 (2015).</li><li>Di Pardo, 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=Impairment+of+blood-brain+barrier+is+an+early+event+in+R6/2+mouse+model+of+Huntington+Disease.">Impairment of blood-brain barrier is an early event in R6/2 mouse model of Huntington Disease.</a> Sci Rep. 7, 41316 (2017).</li><li>Lecler, A., Fournier, L., Diard-Detoeuf, C., Balvay, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood-Brain+Barrier+Leakage+in+Early+Alzheimer+Disease.">Blood-Brain Barrier Leakage in Early Alzheimer Disease.</a> Radiology. 282 (3), 923-925 (2017).</li><li>van de Haar, H. 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=Blood-Brain+Barrier+Leakage+in+Patients+with+Early+Alzheimer+Disease.">Blood-Brain Barrier Leakage in Patients with Early Alzheimer Disease.</a> Radiology. 282 (2), 615 (2017).</li><li>Fernandez-Lopez, 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=Blood-brain+barrier+permeability+is+increased+after+acute+adult+stroke+but+not+neonatal+stroke+in+the+rat.">Blood-brain barrier permeability is increased after acute adult stroke but not neonatal stroke in the rat.</a> J Neurosci. 32 (28), 9588-9600 (2012).</li><li>Yang, Y., Rosenberg, G. 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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplantation+of+pancreatic+islets+into+the+kidney+capsule+of+diabetic+mice.">Transplantation of pancreatic islets into the kidney capsule of diabetic mice.</a> J Vis Exp. (9), e404 (2007).</li><li>Gage, G. J., Kipke, D. R., Shain, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+animal+perfusion+fixation+for+rodents.">Whole animal perfusion fixation for rodents.</a> J Vis Exp. (65), (2012).</li><li>McCloy, R. 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=Partial+inhibition+of+Cdk1+in+G+2+phase+overrides+the+SAC+and+decouples+mitotic+events.">Partial inhibition of Cdk1 in G 2 phase overrides the SAC and decouples mitotic events.</a> Cell Cycle. 13 (9), 1400-1412 (2014).</li><li>Burgess, 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=Loss+of+human+Greatwall+results+in+G2+arrest+and+multiple+mitotic+defects+due+to+deregulation+of+the+cyclin+B-Cdc2/PP2A+balance.">Loss of human Greatwall results in G2 arrest and multiple mitotic defects due to deregulation of the cyclin B-Cdc2/PP2A balance.</a> Proc Natl Acad Sci U S A. 107 (28), 12564-12569 (2010).</li><li>Krueger, M., Hartig, W., Reichenbach, A., Bechmann, I., Michalski, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood-brain+barrier+breakdown+after+embolic+stroke+in+rats+occurs+without+ultrastructural+evidence+for+disrupting+tight+junctions.">Blood-brain barrier breakdown after embolic stroke in rats occurs without ultrastructural evidence for disrupting tight junctions.</a> PLoS One. 8 (2), e56419 (2013).</li><li>Hirano, A., Kawanami, T., Llena, J. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electron+microscopy+of+the+blood-brain+barrier+in+disease.">Electron microscopy of the blood-brain barrier in disease.</a> Microsc Res Tech. 27 (6), 543-556 (1994).</li></ol>

我们在这里描述的技术主要是用来检测脑疾病状态下的 BBB 渗漏。BBB 功能障碍是越来越多的关注, 作为一个常见的特点, 不同的神经系统疾病4。我们以前使用这种方法来描述在一个罕见的神经退行性疾病的小鼠模型, 如 HD6早期紊乱的 BBB 完整性。
该方法利用了程序的相对简单性和有效性, 提供了准确、可靠的结果。它代表了一个高度敏感的?...

An erratum was issued for: <a href="https://www.jove.com/video/56389/assessment-blood-brain-barrier-permeability-intravenous-infusion-fitc">Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease</a>. One of the author&#39;s names was corrected from:
Maglione Vittorio
to:
Vittorio Maglione

An erratum was issued for: <a href="https://www.jove.com/video/56389/assessment-blood-brain-barrier-permeability-intravenous-infusion-fitc">Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease</a>. One of the author's names was corrected.

Erratum: Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease

中枢神经系统内稳态是神经元细胞正常沟通和功能的先决条件。中枢神经系统的实质是由内皮血脑屏障 (BBB) 与周围紧密封闭的, 它代表着外周血和脑的接口, 在这两个区之间的相声中起着举足轻重的作用1 ,2。血脑屏障是一种复杂的动态三维结构, 主要由专门的微血管内皮细胞 (ECs) 通过细胞间连接-紧密接合 (TJs) 和周, 神经元结尾和星形胶质细胞脚处理1,2。
在生理条件下, 完整的血脑屏障的极低通透性确保了对营养物质和其他分子进出大脑的严格调节, 并为中枢神经系统提供了独特的保护, 使其不会发生可能影响神经活动和反对可能的周边侮辱的血液的构成1,2,3。
长期以来, 对 BBB 完整性及其增强的渗透性的破坏已经成为许多神经和神经退行性疾病的关键特征4包括亨廷顿氏病 (HD)5,6, 但是, 这种功能失调是一种致病现象还是在病程中繁殖事件尚不清楚。血脑屏障破裂的时间也仍然难以捉摸, 然而, 我们组和其他人的新证据表明, 扰乱 bbb 完整性并不代表疾病进展的晚期事件, 而是早期的步骤6,7,8, 可能会产生长期后果。
考虑到这一点, 重要的是要早熟揭示 BBB 故障在神经, 以制定战略有用的预测疾病进展和脑损伤, 并制定替代和更有针对性的干预, 能够成功减轻这种破坏的临床后果。因此, 对脑屏障损害的可靠影像学研究, 在大脑疾病的实验和临床管理中具有重要意义。
本文介绍了利用高分子量荧光素异硫氰酸酯标记白蛋白 (FITC-白蛋白) 在 HD 小鼠模型中评价 BBB 通透性的一种成功而简单的方法。FITC-白蛋白的外渗, 通常不能越过屏障, 进入脑实质被测量为 BBB 渗漏的一个指标。该技术易于适应大鼠和其他病理条件的特点, cerebrovasculature 损害9,10。

<table>
	<tbody>
		<tr>
			<td>Albumin-fluorescin isothiocyanate conjiugate</td>
			<td>SIGMA</td>
			<td>A9771-100MG</td>
			<td></td>
		</tr>
		<tr>
			<td>pAb anti-Laminin</td>
			<td>Novus Biologicals</td>
			<td>NB300-144</td>
			<td></td>
		</tr>
		<tr>
			<td>CY3 anti-rabbit made in goat</td>
			<td>MILLIPORE</td>
			<td>AP132</td>
			<td></td>
		</tr>
		<tr>
			<td>SUPERFROST PLUS</td>
			<td>Thermo Scientific</td>
			<td>J1800AMNZ</td>
			<td></td>
		</tr>
		<tr>
			<td>Cover Slips 24 X 50 mm</td>
			<td>Thermo Scientific (DIAPATH)</td>
			<td>61050</td>
			<td></td>
		</tr>
		<tr>
			<td>Kilik Optimal Cutting Temperature (OCT) compound</td>
			<td>Bio Optica</td>
			<td>05-9801</td>
			<td></td>
		</tr>
		<tr>
			<td>VECTASHIELD Mounting Media</td>
			<td>VECTOR</td>
			<td>H-1500</td>
			<td>Mounting media with DAPI</td>
		</tr>
		<tr>
			<td>iNSu/Light Insulin Disposible Syringe</td>
			<td>RAYS Health &#38;Safety</td>
			<td>INS1ML26G13</td>
			<td></td>
		</tr>
		<tr>
			<td>30G 1/2&#34;</td>
			<td>BD Microlance</td>
			<td>304000</td>
			<td>Needle for Insulin disposible Syringe</td>
		</tr>
		<tr>
			<td>Scalpel Handle</td>
			<td>F.S.T.</td>
			<td>91003-12</td>
			<td></td>
		</tr>
		<tr>
			<td>#22 Disposable Scalpel blads</td>
			<td>F.S.T.</td>
			<td>10022-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Iris scissors 10.5 cm</td>
			<td>F.S.T.</td>
			<td>14094-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont Forceps #5745 45&#176; 0.10 x 0.06 mm</td>
			<td>F.S.T.</td>
			<td>11251-35</td>
			<td></td>
		</tr>
		<tr>
			<td>Graefe Forceps 10 cm</td>
			<td>F.S.T.</td>
			<td>11051-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont Forceps #5 0.1 X 0.06 mm</td>
			<td>F.S.T.</td>
			<td>11251-20</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical patch</td>
			<td>Medicalis</td>
			<td>34788</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile disposable towel drape</td>
			<td>Dispotech</td>
			<td>TVO50VE</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereoscopic Microscope</td>
			<td>NIKON</td>
			<td>SMZ 745 T</td>
			<td></td>
		</tr>
		<tr>
			<td>Optic Illuminator LED light (C-FLED2)</td>
			<td>NIKON</td>
			<td>1003167</td>
			<td>Optic Illuminator for Stereoscopic Micrscope</td>
		</tr>
		<tr>
			<td>Eclipse Ni-U Microscope</td>
			<td>Nikon</td>
			<td>932162</td>
			<td>Epifluorescence Microscope</td>
		</tr>
		<tr>
			<td>Microscope digital Camera</td>
			<td>Nikon</td>
			<td>DS-Ri2</td>
			<td>Microscope camera</td>
		</tr>
		<tr>
			<td>Intenslight</td>
			<td>Nikon</td>
			<td>C-HGFI</td>
			<td>Microscope lamp</td>
		</tr>
		<tr>
			<td>NIS-Elements 64 bit</td>
			<td>Nikon</td>
			<td>AR 4.40.00</td>
			<td>Analysis Software</td>
		</tr>
		<tr>
			<td>Electric Razor</td>
			<td>Gemei</td>
			<td>GM-3007</td>
			<td></td>
		</tr>
	</tbody>
</table>

assessment of blood-brain barrier permeability by intravenous infusion of fitc-labeled albumin in a mouse model of neurodegenerative disease

破坏血脑屏障 (BBB) 的完整性是一个常见的特点, 不同的神经和神经退行性疾病。虽然扰动 bbb 稳态和脑疾病的发病机制之间的相互作用需要进一步的研究, 开发和验证可靠的程序, 以准确地检测 bbb 改变可能是至关重要的, 并代表了一个有用的工具潜在的预测疾病进展和制定有针对性的治疗策略。
在这里, 我们提出了一个简单和有效的程序, 评估脑屏障渗漏的神经退行性的情况, 如发生在一个前临床小鼠亨廷顿疾病模型, 其中的缺陷, 血 bbb 的渗透性是明确可探测早熟这种疾病。具体来说, 高分子量的荧光素异硫氰酸酯标记 (FITC)-白蛋白, 只有当后者被削弱时才能够穿过 BBB, 在血管或实质区内被剧烈注入小鼠颈静脉并分布然后由荧光显微镜测定。
绿色荧光-白蛋白在脑实质中的积累是异常 BBB 通透性的一个指标, 当定量使用图像 J 处理软件时, 报告为绿色荧光强度。

适当的 FITC-白蛋白注入颈静脉的结果在绿色荧光示踪剂从血液进入大脑 parenchymawhen BBB 是危及6。在生理条件下, 注入的荧光白蛋白的分布仅限于脑血管内, 周围血管区和/或脑实质的信号都是可探测的 (图 1A, 显微顶)。相反, 当血脑屏障在神经病理环境下被破坏时, FITC-白蛋白在血管空间和大脑实质 (图 1A, 底部显?...

Watch this Scientific Journal Video about Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease at JoVE.com

Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease

在这项研究中, 我们提出了一个简单和有效的程序来评估在神经退行性条件下血脑屏障的破坏。为了达到我们的目标, 我们在小鼠颈静脉内注入了高分子荧光素异硫氰酸酯标记 (FITC)-白蛋白, 并通过荧光显微镜将其渗漏到脑实质中。

FITC 标记白蛋白静脉滴注对神经退行性疾病模型血脑屏障通透性的评价

Disruption of blood-brain barrier (BBB) integrity is a common feature for different neurological and neurodegenerative diseases. Although the interplay ...

The overall goal of this procedure is to assess the brain-blood barrier permeability in a mouse model of degenerative disorder by intrajugular injection of FITC-labeled albumin. This method can help answer the key questions in the study of brain homeostasis. It can be particularly useful to investigate any possible link between blood-brain barrier damage and neurodegeneration.The main advantage of this technique is that it is fast and easy to perform and can be applied, also, to larger animal models and to other neuro-pathological settings. Demonstrating the surgical procedure of intrajugular injection of FITC-albumen will be Luca Capocci. Salvatore Castaldo will demonstrate the histological sectioning of a brains.And Enrico Amico will demonstrate the procedure for micrograph acquisition. Begin by preparing FITC-albumen solution by dissolving FITC-albumen powder in phosphate buffered saline at 10 milligrams per milliliter. Lay out the autoclaved surgical instruments.Clean the surgical bench with 70%ethanol and place a surgical sterile disposable drape. Then set up the operating stereo microscope. After anesthetizing the mouse with an intraperitoneal injection of an appropriate does of ketamine xylazine solution, monitor animal sedation by giving a gentle toe pinch and ensuring the absence of the withdrawal reflex.Place the anesthetized mouse in a face-up supine position and fix the four legs and the tail on the surgical workbench with adhesive tape. After the mouse is secured, carefully shave the appropriate surgical margin on the neck and disinfect it with povidone-iodine solution in 70%ethanol. Dry the exposed shaved surface with cotton swabs.Next, after making a 1.5 centimeter long longitudinal incision in the mid-clavicular line, starting midway on the thorax and extending to just below the neck, carefully stretch apart connective tissue with forceps under microscopic observation to expose the external jugular vein. Ensure sufficient space on the right and on the left of the jugular vein to facilitate the insertion of the 30 and a half gauge needle for the infusion of the FITC-albumen solution. Inject 100 microliters of 10 milligrams per milliliter FITC-albumen in the right jugular vein over three minutes.This is the most critical step during the procedure. Incorrect insertion of the needle can cause irreversible damage of the vein wall and compromise the entire experiment. Fifteen minutes after injection, after euthanizing the mouse by decapitation, rapidly remove the brain from the skull and immerse it in a volume of pre-chilled isopentane, that is ten times that of the brain, for 15 minutes.Move the isopentane frozen brain into a pre-chilled tube and store it in a 80 degrees Celsius freezer until histological sectioning. Embed frozen brain in optimal cutting temperature compound prior to cryostat sectioning. Cut the brain into 20 micron thick sections with a cryostat and mount them onto microscope glass slides.Perform the FITC-albumin laminin co-staining by using a specific anti-laminin antibody, as previously described. Cover slip slides with mounting medium containing DAPI and leave the slide to dry overnight at four degrees Celsius in the dark. The next day, observe the fluorescence staining under a fluorescence microscope at 20 times magnification, equipped with both FITC and Cy3 filters.Turn on the fluorescence lamp about 10 minutes before use. Turn on the microscope, the connected computer and the digital camera. Run the image analysis software.Use the appropriate objective for optimal signal collection and spatial resolution and select appropriate filters. Acquire a minimum of four, 24-bit color exposures per brain slice. Analyze the fluorescence intensity for each single channel, using the freely available ImageJ software.The following representative fluorescence micrographs of brain cryosections show differential distribution of green fluorescent albumin under both physiological and pathological conditions. The red staining, shown here, represents the vascular marker, laminin, under physiological conditions and under conditions of blood-brain barrier compromise. These merged images show co-localization of green fluorescent albumin and laminin, along with DAPI staining of nuclei in the intact and compromised blood-brain barriers.This graph displays the quantification of the green fluorescence emitted by FITC-albumin, which is reported as the green fluorescence intensity in both healthy and compromised conditions. While attempting this procedure, it's important to remember to freshly prepare each single reagent and predispose the surgical bench. After watching this video, you may have a good understanding of how to assess the permeability of the blood-brain barrier in an animal model of neurodegenerative disease, by measuring the affixation of the green fluorescent albumin into brain parenchyma.

Research

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Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

使用超声波血脑屏障破坏和锰增强MRI的脑功能成像

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

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In many neurodegenerative diseases and particularly in Parkinson&#8217;s disease, deficits in olfaction are reported to occur early in the disease process ...

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

用荧光标记示踪剂对小鼠血脑屏障通透性的体内检测

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains ...

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

一种体内评价大鼠缺血性中风模型中血脑屏障的破坏

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

A Human Blood-Brain Interface Model to Study Barrier Crossings by Pathogens or Medicines and Their Interactions with the Brain

研究病原体或药物交叉障碍及其与大脑相互作用的人血脑界面模型

The early screening of nervous system medicines on a pertinent and reliable in cellulo BBB model for their penetration and their interaction with the ...

A Benchtop Approach to the Location Specific Blood Brain Barrier Opening using Focused Ultrasound in a Rat Model

在鼠模型中使用聚焦超声波打开位置特定血脑屏障开口的台面方法

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic ...

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

左旋多巴诱导的帕金森病大鼠模型中运动障碍的诱导与评估

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...

果蝇中的便秘测定

Measuring Constipation in a Drosophila Model of Parkinson's Disease

作者聚焦：探索帕金森病的非运动症状 

在帕金森病的 果蝇 模型中测量便秘

Non-motor symptoms in Parkinson's disease (PD) are common, difficult to treat, and significantly impair quality of life. One prevalent non-motor symptom ...