AbstractAlpha-synuclein (αSyn) forms pathologic aggregates in Parkinson’s disease (PD) and is implicated in mechanisms underlying neurodegeneration. While pathologic αSyn has been extensively studied, there is currently no method to evaluate αSyn within the brains of living patients. Patients with PD are often treated with deep brain stimulation (DBS) surgery in which surgical instruments are in direct contact with neuronal tissue; herein, we describe a method by which tissue is collected from DBS surgical instruments in PD and essential tremor (ET) patients and demonstrate that αSyn is detected.

摘要α-突触核蛋白（αSyn）在帕金森病（PD）中形成病理聚集体，并与神经变性的潜在机制有关。尽管已经对病理性αSyn进行了广泛研究，但目前尚无方法评估活着的患者大脑中的αSyn。PD患者通常接受深部脑刺激（DBS）手术治疗，其中手术器械与神经元组织直接接触；在此，我们描述了一种从PD和原发性震颤（ET）患者的DBS手术器械中收集组织的方法，并证明检测到αSyn。

24 patients undergoing DBS surgery for PD (17 patients) or ET (7 patients) were enrolled; from patient samples, 81.2 ± 44.8 µg of protein (n = 15), on average, was collected from surgical instruments. Light microscopy revealed axons, capillaries, and blood cells as the primary components of purified tissue (n = 3).

纳入24例接受DBS手术治疗PD（17例）或ET（7例）的患者；从患者样本中，平均从手术器械中收集到81.2±44.8µg蛋白质（n=15）。光学显微镜显示轴突，毛细血管和血细胞是纯化组织的主要成分（n=3）。

ELISA assay further confirmed the presence of neuronal and glial tissue in DBS samples (n = 4). Further analysis was conducted using western blot, demonstrating that multiple αSyn antibodies are reactive in PD (n = 5) and ET (n = 3) samples; truncated αSyn (1–125 αSyn) was significantly increased in PD (n = 5) compared to ET (n = 3), in which αSyn misfolding is not expected (0.64 ± 0.25 vs.

ELISA测定进一步证实了DBS样品中存在神经元和神经胶质组织（n=4）。使用蛋白质印迹进行进一步分析，证明多种αSyn抗体在PD（n=5）和ET（n=3）样品中具有反应性；与ET（n=3）相比，PD（n=5）中截短的αSyn（1-125αSyn）显着增加，其中不预期αSyn错误折叠（0.64±0.25 vs。

0.25 ± 0.12, P = 0.046), thus showing that multiple forms of αSyn can be detected from living PD patients with this method..

0.25±0.12，P=0.046），因此表明用这种方法可以从活着的PD患者中检测到多种形式的αSyn。。

IntroductionParkinson’s disease (PD) is a neurodegenerative disorder characterized clinically by a stereotypical movement disorder and neuropathologically by the accumulation of misfolded alpha-synuclein (αSyn) within neurons and glia thought to be etiologic in neuronal dysfunction and symptom manifestation1,2,3,4.

引言帕金森病（PD）是一种神经退行性疾病，临床上表现为刻板运动障碍，神经病理学上表现为神经元和神经胶质内错误折叠的α-突触核蛋白（αSyn）的积累，被认为是神经元功能障碍和症状表现的病因1,2,3,4。

Symptomatically, the disease begins with a prodromal period characterized by dysautonomia and sleep disturbances thought to be consequent to brainstem manifestation of disease, subsequently the progressive movement disorder with bradykinesia and other motor symptoms linked to dopaminergic and striatal dysfunction occur, and eventually cognitive and affective decline traced to alterations in cortical function develops at end stage3,4,5,6.

症状上，该疾病始于前驱期，其特征是自主神经异常和睡眠障碍，被认为是脑干疾病表现的结果，随后出现运动迟缓的进行性运动障碍和与多巴胺能和纹状体功能障碍相关的其他运动症状，最终认知和情感下降可追溯到皮质功能的改变在末期发展3,4,5,6。

Effective treatments for PD mainly exist for alleviation of motor symptoms, with dopaminergic augmentation therapies such as levodopa and similar pharmaceuticals being first line treatment after diagnosis which occurs after classical movement disorder symptoms are evident4. When motor symptoms become refractory to medical management, or dyskinetic side effects from these medications are intolerable, neurosurgical treatment with deep brain stimulation (DBS) to modulate striatal circuitry involved in voluntary movement has proven to be an effective treatment for bradykinesia, tremor, rigidity, and dyskinesias4,7.

PD的有效治疗主要是为了缓解运动症状，多巴胺能增强疗法如左旋多巴和类似药物是诊断后的一线治疗，这种治疗发生在经典运动障碍症状明显后4。当运动症状变得难以治疗，或者这些药物的运动障碍副作用无法忍受时，用深部脑刺激（DBS）调节参与自主运动的纹状体回路的神经外科治疗已被证明是治疗运动迟缓，震颤，僵硬和运动障碍的有效方法4,7。

Similarly, essential tremor (ET) is frequently treated with DBS to modulate movement circuitry through the dentato-rubro-thalamic tract (DRTT) and ventral intermediate thalamic nucleus (VIM); PD and ET differ in neuropathology in that misfolded αSyn inclusions are not abundant or causally implicated in ET7,8.

同样，经常用DBS治疗原发性震颤（ET），以通过齿状红丘脑束（DRTT）和腹侧丘脑中间核（VIM）调节运动回路；。

Dopaminergic medications and DBS provide substantial relief of impaired motor function in PD, however, there cu.

多巴胺能药物和DBS可以显着缓解PD中运动功能受损的情况，但是，有cu。

Data availability

数据可用性

Data is available upon request from the corresponding author (Zachary Sorrentino) to aid in interpretation and verification of research contained herein. The authors are open to collaborations for further investigations using this method.

应通讯作者（扎卡里·索伦蒂诺）的要求，可以获得数据，以帮助解释和验证本文所载的研究。作者对使用这种方法进行进一步研究的合作持开放态度。

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Cariulo，C。等人。通过超灵敏免疫测定法测定，磷酸S129α-突触核蛋白存在于人血浆中，但不存在于脑脊液中。正面。神经科学。13889年。https://doi.org/10.3389/fnins.2019.00889（2019年）。文章

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Download referencesAcknowledgementsThe authors would like to thank the University of Florida Center for Translational Research in Neurodegenerative Disease for access to lab space, common items, and facilitation of research. The authors would also like to thank all research participants for volunteering in this study.FundingThis work was supported by grants from the National Institutes of Health (RF1NS129567, R01NS100876) and internal funds from the University of Florida Department of Neurosurgery.

下载参考文献致谢作者要感谢佛罗里达大学神经退行性疾病转化研究中心访问实验室空间，常见项目和促进研究。作者还要感谢所有参与这项研究的志愿者。资助这项工作得到了美国国立卫生研究院（RF1NS129567，R01NS100876）的资助和佛罗里达大学神经外科的内部资金的支持。

G.M.L and S.Q. were supported by T32NS082168 training grant from National Institute of Neurological Disorders and Stroke.Author informationAuthors and AffiliationsUniversity of Florida College of Medicine, 1505 SW Archer Rd, Gainesville, FL, 32608, USAZachary A. Sorrentino, Joshua Riklan, Grace M. Lloyd, Brandon P.

G、 M.L和S.Q.得到了美国国家神经疾病和中风研究所的T32NS082168培训资助。作者信息作者和附属机构佛罗里达大学医学院，1505 SW Archer Rd，盖恩斯维尔，佛罗里达州，32608，USAZachary A.Sorrentino，Joshua Riklan，Grace M.Lloyd，Brandon P。

Lucke-Wold, David Mampre, Stephan Quintin, Rasheedat Zakare-Fagbamila, Megan Still, Vyshak Chandra, Kelly D. Foote, Benoit I. Giasson & Justin D. HilliardDepartment of Neurosurgery, University of Florida College of Medicine, Gainesville, FL, USAZachary A. Sorrentino, Brandon P. Lucke-Wold, David Mampre, Rasheedat Zakare-Fagbamila, Megan Still, Vyshak Chandra, Kelly D.

Lucke Wold，David Mampre，Stephan Quintin，Rasheedat Zakare Fagbamila，Megan Still，Vyshak Chandra，Kelly D.Foote，Benoit I.Giasson＆Justin D.HilliardDepartment of Neurosurgery，佛罗里达大学医学院，佛罗里达州盖恩斯维尔，USAZachary A.Sorrentino，Brandon P.Lucke Wold，David Mampre，Rasheedat Zakare Fagbamila，Megan Still，Vyshak Chandra，Kelly D。

Foote & Justin D. HilliardDepartment of Neuroscience, University of Florida College of Medicine, Gainesville, FL, USAGrace M. Lloyd, Stephan Quintin & Benoit I. GiassonAuthorsZachary A. SorrentinoView author publicationsYou can also search for this author in.

Foote＆Justin D.HilliardDepartment of Neuroscience，University of Florida College of Medicine，盖恩斯维尔，佛罗里达州，USAGRES M.Lloyd，Stephan Quintin＆Benoit I.GiassonAuthorsZachary A.SorrentinoView author Publications您也可以在中搜索这位作者。

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PubMed Google ScholarContributionsZ.S. participated in conception, design, regulatory approval, data collection, analysis, manuscript writing and editing. J.R., G.L. participated in data analysis and manuscript writing and editing. B.L., S.Q., D.M., R.Z., M.S., V.C., K.F. participated in data collection and manuscript editing.

PubMed谷歌学术贡献。S、 参与构思，设计，监管批准，数据收集，分析，稿件撰写和编辑。J、 R.，G.L.参与了数据分析以及手稿的撰写和编辑。B、 L.，S.Q.，D.M.，R.Z.，M.S.，V.C.，K.F.参与了数据收集和手稿编辑。

B.G. and J.H. participated in conception, design, regulatory approval, data collection, manuscript writing and editing.Corresponding authorCorrespondence to.

B、 G.和J.H.参与了构思，设计，监管批准，数据收集，手稿撰写和编辑。对应作者对应。

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Reprints and permissionsAbout this articleCite this articleSorrentino, Z.A., Riklan, J., Lloyd, G.M. et al. Neuronal tissue collection from intra-cranial instruments used in deep brain stimulation surgery for Parkinson’s disease with implications for study of alpha-synuclein.

转载和许可本文引用本文Sorrentino，Z.A.，Riklan，J.，Lloyd，G.M.等人从用于帕金森氏病深部脑刺激手术的颅内仪器中收集神经元组织，对α-突触核蛋白的研究有影响。

Sci Rep 14, 21641 (2024). https://doi.org/10.1038/s41598-024-72542-5Download citationReceived: 04 May 2024Accepted: 09 September 2024Published: 16 September 2024DOI: https://doi.org/10.1038/s41598-024-72542-5Share this articleAnyone you share the following link with will be able to read this content:Get shareable linkSorry, a shareable link is not currently available for this article.Copy to clipboard.

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KeywordsParkinson’s diseaseAlpha-synucleinEssential tremorDeep brain stimulationNeurodegeneration

关键词Sparkinson病α-突触核必需性震颤深部脑刺激神经变性

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BiomarkersCytological techniquesHistologyMedical researchMovement disordersNeurodegenerative diseasesNeurological disordersParkinson's diseaseTranslational research

生物标志物细胞学技术医学研究运动障碍神经退行性疾病神经系统疾病Sparkinson疾病转化研究

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