科研助理

刘宇、孔维贤

研究生

艾璞、刘倚含、周林林、李成杰、王宇炜、王均鑫

博士后

陈慧丽

本科生

李瑞妍（国豪书院）

合作学生

胡瑞芹、宿可

国家自然科学基金委员会面上项目、青年项目,  上海市“科技创新行动计划”自然科学基金面上项目, 宁夏回族自治区科技厅自治区重点研发计划对外科技合作项目, 上海市“浦江人才”项目，上海市第四人民医院启动经费，同济大学“青年百人”计划启动经费等。

课题组通过构建全新膜片钳-电化学偶联的双神经递质检测平台、长时程树突棘动力学成像、海马和中脑脑片培养等前沿技术，率先阐明了非遗传因素神经元活性（neuronal activity）调控α-syn聚集并加速其脑区传播的分子机制，深入研究了多巴胺与多种单胺类神经递质“量子化”释放的调控机制，解析了多巴胺能神经元易感性的致病机理，得到了国际同行的证实与广泛关注（图1）。 

                                                                                        图1：课题组总体研究示意图

（1）构建全新脑片培养系统，解析神经元活性调控a-syn聚集和传播新机制

课题组优化了小鼠海马和中脑脑片培养系统，结合α-syn PFF，借助化学遗传学提升神经元活性，发现α-syn聚集体在海马脑片上显著增加，在中脑脑片上伴随更多路易小体和多巴胺能神经元的死亡，并加速α-syn聚集体在小鼠不同脑区传播（Acta Neuropathologica，2020，第一作者）。在原代培养的海马神经元上，α-syn PFF抑制神经元mEPSCs发放频率，突触前膜和后膜中都有α-syn聚集体堆积，且显著抑制新生突触的形成（Journal of Neuroscience，2019，第一作者；Frontiers in Neuroscience，2021，通讯作者）（图2）。    

                                                                                   图2：解析α-synuclein的病理、生理功能。

（2）建立膜片钳-电化学偶联检测系统，解析单胺类递质“量子化”释放新机制

课题组使用电化学、荧光成像等技术发现，去甲肾上腺素的分泌量在动力蛋白Dynamin 1敲除的嗜铬细胞上增加200 %，并且伴随着囊泡融合小孔开放直径的增加（Journal of Neuroscience，2019，第一作者），修正了Katz等人提出的“量子化”分泌理论。同时，我们创新性的构建了Patch sniffer技术发现，其中ATP处于“量子化”释放，而去甲肾上腺素处于“亚量子化”释放，且主要受到囊泡内基质蛋白（Chromogranin A）的调控（Neuron，2019，共同第一作者）。此外，在背根神经节DRG神经元上，电压门控钙离子通道VGCCs通过介导瞬时性钙离子内流，在轴突上引起“量子化”释放；而配体门控钙离子通道TRPV1则通过介导持续性钙离子内流，在胞体里引发“亚量子化”释放模式（Science Signaling，2019，共同第一作者，封面文章，Science重点推介）（图3）。      

                                                                           图3：解析单胺类神经递质释放的调控机制。

Ø  Jiang J, Liu Y, Wu Q#. Revisit the Cellular Transmission and Emerging Techniques in Understanding the Mechanisms of Proteinopathies. Front Neurosci. 2021 Nov 18;15:781722. doi: 10.3389/fnins.2021.781722.eCollection 2021. (# Corresponding author)

Ø  Wu Q, Shaik M, Meymand E, Zhang B, Luk K, Trojanowski JQ, Lee VM. Neuronal activity modulates alpha‐synuclein aggregation and spreading in organotypic brain slice cultures and in vivo. Acta Neuropathol. 2020 Oct 6. doi: 10.1007/s00401-020-02227-6. Online ahead of print. 

Ø  Wu Q, Takano H, Riddle DM, Trojanowski JQ, Coulter DA, Lee VM. α-Synuclein (αSyn) Preformed Fibrils Induce Endogenous αSyn Aggregation, Compromise Synaptic Activity and Enhance Synapse Loss in Cultured Excitatory Hippocampal Neurons. J Neurosci. 2019 Jun 26;39(26):5080-5094.

Ø  Zhang Q*, Liu B*, Wu Q*, Liu B, Li Y, Sun S, Wang Y, Wu X, Chai Z, Jiang X, Liu X, Hu M, Wang Y, Yang Y, Wang L, Kang X, Xiong Y, Zhou Y, Chen X, Zheng L, Zhang B, Wang C, Zhu F, Zhou Z. Differential Co-release of Two Neurotransmitters from a Vesicle Fusion Pore in Mammalian Adrenal Chromaffin Cells. Neuron. 2019 Apr 3;102(1):173-183.e4. (* Co-first author)

Ø  Wu Q*, Zhang Q*, Liu B, Li Y, Wu X, Kuo S, Zheng L, Wang C, Zhu F, Zhou Z. Dynamin 1 Restrains Vesicular Release to a Subquantal Mode in Mammalian Adrenal Chromaffin Cells. J Neurosci. 2019 Jan 9;39(2):199-211. 

Ø  Wang Y*, Wu Q*, Hu M, Liu B, Chai Z, Huang R, Wang Y, Xu H, Zhou L, Zheng L, Wang C, Zhou Z. Ligand- and voltage-gated Ca2+ channels differentially regulate the mode of vesicular neuropeptide release in mammalian sensory neurons. Sci Signal. 2017 Jun 20;10(484). (* Co-first author, Front Cover).

Highlighted in Science (Ferrarelli LK, 2017, Kiss-and-run or a full commitment? 356(6344):1243-1245. doi: 10.1126/science.356.6344.1243-p). 

Ø  Kuo S*, Wu Q*, Liu B*, Xie Z, Wu X, Shang S, Zhang X, Zhou Z. Importing, caring, breeding, genotyping, and phenotyping a genetic mouse in a Chinese university. J Mol Neurosci. 2014 Jul;53(3):487-92. (* Co-first author)

Ø  Ji P, An B, Jie Z, Wang L, Qiu S, Ge C, Wu Q, Shi J, Huo M. Genetically engineered probiotics as catalytic glucose depriver for tumor starvation therapy. Mater Today Bio. 2022 Dec 15;18:100515.  doi: 10.1016/j.mtbio.2022.100515. eCollection 2023 Feb.

Ø  Zhou H, Jiao B, Ouyang Z, Wu Q, Shen L, Fang L. Report of two pedigrees with heterozygous HTRA1 variants-related cerebral small vessel disease and literature review. Mol Genet Genomic Med. 2022 Oct;10(10):e2032.  doi: 10.1002/mgg3.2032. Epub 2022 Aug 10.

Ø  Bassil F, Meymand E, Brown H, Xu H, Cox T, Pattabhiraman S, Maghames C, Wu Q, Zhang B, Trojanowski J, Lee M. α-Synuclein modulates tau spreading in mouse brains. J Exp Med. 2021 Jan 4;218(1):e20192193. doi: 10.1084/jem.20192193.  

Ø  Gibbons G, Kim SJ, Wu Q, Riddle DM, Leight SN, Changolkar L, Xu H, Meymand ES, O'Reilly M, Zhang B, Brunden KR, Trojanowski JQ, Lee VM.  Conformation-selective tau monoclonal antibodies inhibit tau pathology in primary neurons and a mouse model of Alzheimer's disease. Molecular Neurodegeneration. 2020 Nov 4;15(1):64. doi: 10.1186/s13024-020-00404-5.

Ø  Zhang Q, Liu B, Li Y, Yin L, Younus M, Jiang X, Lin Z, Sun S, Huang R, Liu B, Wu Q, Zhu F, Zhou Z. Regulating quantal size of neurotransmitter release through a GPCR voltage sensor. Proc Natl Acad Sci U S A. 2020 Oct 27;117(43):26985-26995. doi: 10.1073/pnas.2005274117. Epub 2020 Oct 12. 

Ø  Wang C, Kang X, Zhou L, Chai Z, Wu Q, Huang R, Xu H, Hu M, Sun X, Sun S, Li J, Jiao R, Zuo P, Zheng L, Yue Z, Zhou Z. Synaptotagmin-11 is a critical mediator of parkin-linked neurotoxicity and Parkinson's disease-like pathology. Nat Commun. 2018 Jan 8;9(1):81.

Ø  Chai Z, Wang C, Huang R, Wang Y, Zhang X, Wu Q, Wang Y, Wu X, Zheng L, Zhang C, Guo W, Xiong W, Ding J, Zhu F, Zhou Z. CaV2.2 Gates Calcium-Independent but Voltage-Dependent Secretion in Mammalian Sensory Neurons. Neuron. 2017 Dec 20;96(6):1317-1326.e4. 

Ø  Zhu F, Wu Q, Li J, Grycel K, Liu B, Sun X, Zhou L, Jiao R, Song R, Khan YM, Wang Q, Wang L, Xu Y, Li J, Zhang B, Zhou Z. A single dose of cocaine potentiates glutamatergic synaptic transmission onto locus coeruleus neurons. Cell Calcium. 2017 Nov;67:11-20. 

Ø  Fan H, Du X, Zhang J, Zheng H, Lu X, Wu Q, Li H, Wang H, Shi Y, Gao G, Zhou Z, Tan DX, Li X.  Selective inhibition of Ebola entry with selective estrogen receptor modulators by disrupting the endolysosomal calcium. Sci Rep. 2017 Jan 24;7:41226. 

Ø  Shang S, Zhu F, Liu B, Chai Z, Wu Q, Hu M, Wang Y, Huang R, Zhang X, Wu X, Sun L, Wang Y, Wang L, Xu H, Teng S, Liu B, Zheng L, Zhang C, Zhang F, Feng X, Zhu D, Wang C, Liu T, Zhu MX, Zhou Z. Intracellular TRPA1 mediates Ca2+ release from lysosomes in dorsal root ganglion neurons. J Cell Biol. 2016 Nov 7;215(3):369-381. 

Ø  Wang C, Wang Y, Hu M, Chai Z, Wu Q, Huang R, Han W, Zhang CX, Zhou Z. Synaptotagmin-11 inhibits clathrin-mediated and bulk endocytosis. EMBO Rep. 2016 Jan;17(1):47-63.

Ø  Xu H, Zuo P, Wang S, Zhou L, Sun X, Hu M, Liu B, Wu Q, Dou H, Liu B, Zhu F, Teng S, Zhang X, Wang L, Li Q, Jin M, Kang X, Xiong W, Wang C, Zhou Z. Striatal dopamine release in a schizophrenia mouse model measured by electrochemical amperometry in vivo. Analyst. 2015 Jun 7;140(11):3840-5. 

Ø  Dou H, Wang C, Wu X, Yao L, Zhang X, Teng S, Xu H, Liu B, Wu Q, Zhang Q, Hu M, Wang Y, Wang L, Wu Y, Shang S, Kang X, Zheng L, Zhang J, Raoux M, Lang J, Li Q, Su J, Yu X, Chen L, Zhou Z. Calcium influx activates adenylyl cyclase 8 for sustained insulin secretion in rat pancreatic beta cells. Diabetologia. 2015 Feb;58(2):324-33. 

Ø  Kang X, Xu H, Teng S, Zhang X, Deng Z, Zhou L, Zuo P, Liu B, Liu B, Wu Q, Wang L, Hu M, Dou H, Liu W, Zhu F, Li Q, Guo S, Gu J, Lei Q, Lü J, Mu Y, Jin M, Wang S, Jiang W, Liu K, Wang C, Li W, Zhang K, Zhou Z. Dopamine release from transplanted neural stem cells in Parkinsonian rat striatum in vivo. Proc Natl Acad Sci U S A. 2014 Nov 4;111(44):15804-9. 

Ø  Shang S, Wang C, Liu B, Wu Q, Zhang Q, Liu W, Zheng L, Xu H, Kang X, Zhang X, Wang Y, Zheng H, Wang S, Xiong W, Liu T, Zhou Z. Extracellular Ca²⁺ per se inhibits quantal size of catecholamine release in adrenal slice chromaffin cells. Cell Calcium. 2014 Sep;56(3):202-7. 

Ø  Wang L, Zhang X, Xu H, Zhou L, Jiao R, Liu W, Zhu F, Kang X, Liu B, Teng S, Wu Q, Li M, Dou H, Zuo P, Wang C, Wang S, Zhou Z. Temporal components of cholinergic terminal to dopaminergic terminal transmission in dorsal striatum slices of mice. J Physiol. 2014 Aug 15;592(16):3559-76. 

Ø  Wang L, Shang S, Kang X, Teng S, Zhu F, Liu B, Wu Q, Li M, Liu W, Xu H, Zhou L, Jiao R, Dou H, Zuo P, Zhang X, Zheng L, Wang S, Wang C, Zhou Z. Modulation of dopamine release in the striatum by physiologically relevant levels of nicotine. Nat Commun. 2014 May 21;5:3925. 

Ø  Zheng L, Wang C, Shang S, Zhang X, Wang Y, Wu Q, Hu M, Chai Z, Wu X, Zheng H, Zhang C, Wang L, Xiong W, Zhou Z. Real-time endocytosis imaging as a rapid assay of ligand-GPCR binding in single cells. Am J Physiol Cell Physiol. 2013 Oct 1;305(7):C751-60. 

Ø  Zhang B, Zhang X, Luo P, Huang W, Zhu F, Liu T, Du Y, Wu Q, Lü J, Xiu Y, Liu L, Huang H, Guo S, Zheng H, Zhang C, Zhou Z. Action potential-triggered somatic exocytosis in mesencephalic trigeminal nucleus neurons in rat brain slices. J Physiol. 2012 Feb 15;590(4):753-62.