Journal of Shandong University (Health Sciences) ›› 2020, Vol. 58 ›› Issue (3): 65-74.doi: 10.6040/j.issn.1671-7554.0.2019.1369

Previous Articles     Next Articles

Application of adeno-associated virus on thalamic striatum pathway in rats

HE Tianqi1,2, LI Min3, WANG Xuenan3, WANG Yanan3, LI Yuchuan3, SUN Shuang3, ZHAO Hailong1,2, WANG Hao2, CHEN Dadian3, ZHU Meijia2, WANG Xiaojun2, WANG Min3, LI Xiuhua2   

  1. 1. Weifang Medical University, Weifang 261053, Shandong, China;
    2.Department of Neurology, The First Affiliated Hospital of Shandong First Medical University, Jinan 250014, Shandong, China;
    3. Key Laboratory of Animal Resistance Biology of Shandong Province, College of Life Science, Shandong Normal University, Jinan 250014, Shandong, China
  • Online:2020-03-10 Published:2022-09-27

PDF (PC)

44

Abstract: Objective To investigate the neuronal projection of adeno-associated virus(AAV2/9-GFP)in parafascicular nucleus-striatum pathway and its effect on tissues. Methods A total of 136 adult male Wistar rats were randomly divided into parafascicular nucleus(PF)group (n=64), dorsolateral striatum(DLS)group(n=64), PF control group(n=4)and DLS control group (n=4)to conduct stereotactic localization experiments. Based on stereotactic localization experiments, behavior experiments were performed in PF behavior group(n=12)and behavior control group(n=4). After stereotactic localization experiments, the neuron counting experiments were conducted in PF counting group (n=16), 山 东 大 学 学 报 (医 学 版)58卷3期 -何天齐,等.腺相关病毒在大鼠丘脑纹状体通路中的应用 \=- DLS counting group(n=16), PF counting control group(n=4)and DLS counting control group(n=4). After different doses of AAV2/9-GFP were injected into the PF group and DLS group for different durations, the transfection efficiency was analyzed, and the optimal dose and transfection period were screened. The transfected neurons and neuronal projection pathways between the two nuclei were observed. The behavioral changes of rats and number and morphology of neurons were analyzed. SPSS Statistics 21.0 software was used for statistical analysis. Results With the increase of transfection doses and time, the PF optical density value increased (Fdoses=55.617, P<0.001; Ftime=65.288, P<0.001), and the transfection area increased (Fdoses=202.443, P<0.001; Ftime=224.064, P<0.001), showing an upward trend. With the increase of transfection doses and time, the DLS optical density value increased (Fdoses=117.062, P<0.001; Ftime=155.792, P<0.001), and transfection area increased (Fdoses=240.422, P<0.001; Ftime=304.571, P<0.001), showing an upward trend. The optimal dose of PF for viral transfection was 0.4 μL and the optimal duration was 4 weeks. The optimal dose of DLS transfection was 0.6 μL and the optimal duration was 4 weeks. When AAV2/9-GFP was injected into PF, it was projected to the dorsolateral striatum, motor cortex and insular lobe. When AAV2/9-GFP was injected to DLS, it was projected to the motor cortex, hypothalamic nucleus, parafascicular nucleus and substantia nigra. AAV2/9-GFP had damage to neurons, which was characterized by a decreased number of neurons (P<0.05), irregular morphology, enlarged cell body, and shallower staining of nucleoli and Nissl bodies. Conclusion There is a close neuronal projection relationship between PF, DLS, motor cortex, hypothalamic nucleus and substantia nigra. AAV2/9-GFP is an effective neural tracer and harmful to neurons, so it should be cautiously used in clinical practice.

Key words: Adeno-associated virus, Parafascicular nucleus, Dorsolateral striatum, Pathway, Rat

CLC Number: 

  • R741
[1] Womelsdorf T, Valiante TA, Sahin NT, et al. Dynamic circuit motifs underlying rhythmic gain control, gating and integration [J]. Nat Neurosci, 2014, 17(8): 1031-1039.
[2] Blessing D, Déglon N. Adeno-associated virus and lentivirus vectors: a refined toolkit for the central nervous system [J]. Curr Opin Virol, 2016, 21: 61-66. doi: 10.1016/j.coviro.2016.08.004.
[3] Hocquemiller M, Giersch L, Audrain M, et al. Adeno-associated virus-based gene therapy for CNS diseases [J]. Hum Gene Ther, 2016, 27(7): 478-496.
[4] 周斌, 胡镐申, 陈玲敏, 等. 腺相关病毒介导的小鼠发育期大脑中的特定细胞和特定脑区的基因编辑及潜在应用探索[J]. 临床和实验医学杂志, 2019, 18(3):225-229. ZHOU Bin, HU Haoshen, CHEN Lingmin, et al. Adeno-associated virus mediated cellular and regional specific gene editing and the potential applications in the developmental brain of mice [J]. Journal of Clinical and Experimental Medicine, 2019, 18(3): 225-229.
[5] 李冬晓, 刘崇, 郭艳苏. 腺相关病毒在中枢神经系统的转导与表达特性[J]. 脑与神经疾病杂志, 2016, 24(5): 322-327.
[6] Chan KY, Jang MJ, Yoo BB, et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems [J]. Nature Neurosci, 2017, 20(8): 1172-1179.
[7] Galvan A, Devergnas A, Wichmann T. Alterations in neuronal activity in basal ganglia-thalamocortical circuits in the parkinsonian state [J]. Front Neuroanat, 2015, 9: 5. doi: 10.3389/fnana.2015.00005.
[8] Calabresi P, Picconi B, Tozzi A, et al. Direct and indirect pathways of basal ganglia: a critical reappraisal [J]. Nat Neurosci, 2014, 17(8): 1022-1030.
[9] Cowan WM, Powell TP. A study of thalamo-striate relations in the monkey [J]. Brain, 1956, 79(2): 364-390.
[10] Villalba RM, Wichmann T, Smith Y. Neuronal loss in the caudal intralaminar thalamic nuclei in a primate model of Parkinsons disease [J]. Brain Struct Funct, 2014, 219(1): 381-394.
[11] Kusnoor SV, Bubser M, Deutch AY. The effects of nigrostriatal dopamine depletion on the thalamic parafascicular nucleus [J]. Brain Res, 2012, 1446: 46-55. doi: 10.1016/j.brainres.2012.01.040.
[12] Parent M, Parent A. Single-axon tracing and three-dimensional reconstruction of centre median-parafascicular thalamic neurons in primates [J]. J Comp Neurol, 2005, 481(1): 127-144.
[13] Paxinos G, Watson CR, Emson PC. AChE-stained horizontal sections of the rat brain in stereotaxic coordinates [J]. J Neurosci Methods, 1980, 3(2): 129-149.
[14] Murlidharan G, Samulski RJ, Asokan A. Biology of adeno-associated viral vectors in the central nervous system [J]. Front Mol Neurosci, 2014, 7: 76. doi: 10.3389/fnmol.2014.00076.
[15] Ojala DS, Amara DP, Schaffer DV. Adeno-associated virus vectors and neurological gene therapy [J]. Neuroscientist, 2015, 21(1): 84-98.
[16] Naldini, Luigi. Gene therapy returns to centre stage [J]. Nature, 2015, 526(7573): 351-360.
[17] Tervo DG, Hwang BY, Viswanathan S, et al. A designer AAV variant permits efficient retrograde access to projection neurons [J]. Neuron, 2016, 92(2): 372-382.
[18] Gray SJ, Matagne V, Bachaboina L, et al. Preclinical differences of intravascular AAV9 delivery to neurons and glia: A comparative study of adult mice and nonhuman primates [J]. Mol Ther, 2011, 19(6): 1058-1069.
[19] Samaranch L, Salegio EA, San Sebastian W, et al. Adeno-associated virus serotype 9 transduction in the central nervous system of nonhuman primates [J]. Hum Gene Ther, 2013, 24(5): 526-532.
[20] 陆元喜. 重组9型腺相关病毒载体通过不同方法转染SD大鼠心肌转染效率及安全性研究[D]. 南宁: 广西医科大学, 2017.
[21] Zhang H, Yang J, Wang X, et al. Altered local field potential relationship between the parafascicular thalamic nucleus and dorsal striatum in hemiparkinsonian rats [J]. Neurosci Bull, 2019, 35(2): 315-324.
[22] Zhang H, Yang J, Xiang T, et al. State-dependent spike and local field synchronization between the thalamic parafascicular nucleus and the dorsal striatum in a rat model of Parkinsons disease [J]. Neuroscience, 2019, 404: 27-38. doi: 10.1016/j.neuroscience.2019.01.055.
[23] Parent M, Parent A. Single-axon tracing and three-dimensional reconstruction of centre médian-parafascicular thalamic neurons in primates [J]. J Comp Neurol, 2005, 481(1): 127-144.
[24] Galvan A, Smith Y. The primate thalamostriatal systems: Anatomical organization, functional roles and possible involvement in Parkinsons disease [J]. Basal Ganglia, 2011, 1(4): 179-189.
[25] Samaranch L, Sebastian WS, Kells AP, et al. AAV9-mediated expression of a non-self protein in nonhuman primate central nervous system triggers widespread neuroinflammation driven by antigen-presenting cell transduction [J]. Mol Ther, 2014, 22(2): 329-337.
[26] Löw K, Aebischer P, Schneider BL.Direct and retrograde transduction of nigral neurons with AAV6, 8, and 9 and intraneuronal persistence of viral particles [J]. Hum Gene Ther, 2013, 24(6): 613-629.
[1] YU De-Xin, CAEI Ti-Gong, MA Xiang-Xing, ZHANG Xiao-Ming, LI Chuan-Fu. Angiogenesis and maturation of hepatocellular carcinoma [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 51-54.
[2] WUYi-Na, GE Zhi-Ming, LI Fang, HE Hong, JIANG Hong, ZHANG Yun. Expressions of angiotensin converting enzyme 2 mRNA and protein in the kidneys in spontaneously hypertensive rats [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 5-.
[3] ZHU Lin, HU San-Yuan, ZHANG Guang-Yong, DING Xiang-Jiu. Protection of PGE2 against intestinal mucosa injuries in obstructed jaundice rats [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 12-.
[4] SUN Tao, ZHANG Dao-Lai, XIE Shan-Shan, WANG Yu-Zhuo, FENG Yu-Xin, XIN Hua. Effect of ethanol on expression of connexin 43 in primary culture of neural precursor cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 20-.
[5] ZHANG Dao-Lai, SUN Tao, XIE Shan-Shan, WANG Yu-Zhuo, ZHAO Ling, FENG Yu-Xin, XIN Hua. Developmental expression of NMDAR1 subunit on cultured cerebral cortical neurons of fetal rats in vitro [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 28-32.
[6] WANG Xiao-Lei, ZHANG Hai-Thao, ZHANG Hui, GUO Cheng-Hao. Protective effects of shuxuening injecta on vascular  endothelial  cell injury in vitro due to iodine excesses [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 38-.
[7] LU Xiang-Dong, YANG Wei, XU Guang-Meng, QU Yuan-Ming. Expression and role of PPAR-γin meningiomas and troglitazone induced meningiomas cell apoptosis in vitro [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 65-.
[8] ZHANG Xiao-Yan, LI Gang, WANG Ling-Beng. Effects of different dosages of Sufentanil in induced general anesthesia  period on blood rheology cortisol in spinal operations [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 76-79.
[9] WANG Xin, XING Chun-Yan, YANG Yan-Ping. Serum phosphopyruvate hydratase in diagnosis of invasive Candida albicans infection [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 92-94.
[10] LU Qun, ZHAO Lulu. Progress on assisted reproductive technology for women with atypical endometrial hyperplasia and endometrial carcinoma receiving fertilitysparing treatment [J]. Journal of Shandong University (Health Sciences), 2022, 60(9): 35-41.
[11] ZHAO Ge, ZOU Cunhua, SONG Dongdong, ZHAO Shuping. Effects of tanshinone IIA on the proliferation and apoptosis of endometrial carcinoma cells [J]. Journal of Shandong University (Health Sciences), 2022, 60(9): 53-58.
[12] ZOU Pinheng, CHEN Tianguo, HU Kang, LI Weicai. Effects and mechanism of overexpression of miR-27a on hippocampal neuronal injury in rats with acute cerebral infarction [J]. Journal of Shandong University (Health Sciences), 2022, 60(9): 59-66.
[13] ZHANG Zhenwei, LI Jia, CHEN Keming. IGF2BP2/m6A/ITGA5 signal axis regulates the proliferation and migration of renal clear cells [J]. Journal of Shandong University (Health Sciences), 2022, 60(9): 74-84.
[14] HAN Jing, JIA Chunling. Effects of periodontal basic therapy on postoperative pneumonia in patients with lung cancer before thoracic surgery [J]. Journal of Shandong University (Health Sciences), 2022, 60(9): 113-118.
[15] FU Jia, WANG Lulu, HU Shuai, CHEN Zheping, LIU Dongyi, LI Qingsong, LU Guodong, ZHANG He, ZHAO Xin, FENG Chang. Effects of different approaches of quadrates lumborum block on postoperative recovery of patients undergoing radical gastrectomy [J]. Journal of Shandong University (Health Sciences), 2022, 60(8): 50-57.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd