«上一篇/Previous Article|本期目录/Table of Contents|下一篇/Next Article»

《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

26卷

期数:

2020年01期

页码:

1-7

栏目:

基础医学

出版日期:

2020-04-06

文章信息/Info

Title:

Establishment and identification of TauT transgenic rats

文章编号:

1006-8147(2020)01-0001-07

作者:

齐浩铭¹; 仝慧慧¹; 范维佳²; 王 辰²; 莫丽冬²; 徐立霞²; 么秀华²; 黄慧玲²

（1.天津医科大学研究生院，天津300070；2.南开大学附属环湖医院，天津市神经外科研究所，天津市脑血管与神经变性重点实验室，天津300350）

Author(s):

QI Hao-ming¹;  TONG Hui-hui¹;  FAN Wei-jia²;  WANG Chen²;  MO Li-dong²;  XU Li-xia²;  YAO Xiu-hua²;  HUANG Hui-ling²

（1.Gradute School, Tianjin Medical University, Tianjin 300070, China； 2.Tianjin Huanhu Hospital, Tianjin Neurosurgical Institute, Tianjin Key Laboratory of Cerebrovascular and Neurodegenerative Diseases, Tianjin 300350, China)

关键词:

牛磺酸转运体; 大鼠; 转基因

Keywords:

taurine transporter;  rat;  knock in

分类号:

R332

DOI:

-

文献标志码:

A

摘要:

目的：为研究牛磺酸转运体（TauT）在神经系统疾病中的作用，构建并繁育脑特异性表达TauT转基因模型大鼠（TauT+/-大鼠）。方法：将TAUT cDNA插入脑特异性PDGF启动子下游，构建转基因表达载体，显微注射法建立TAUT转基因大鼠。Genotype鉴定转基因大鼠的基因型。经过配种繁殖和鉴定后，绘制2～8月龄生长曲线、测定各脏器质量、脏体系数和脏脑系数；RT-PCR技术检测TauT+/-大鼠大脑组织中Slc6a6 mRNA基因表达水平；Western blot 检测TauT+/-大鼠在大脑、心、肝组织的TauT蛋白表达，并对TauT+/-大鼠大脑组织进行免疫组化染色和HE染色，IPP软件对免疫组化图像进行处理。结果：成功构建PDGF-Slc6a6表达质粒，得到3只TauT+/-大鼠F0代。经过繁殖和鉴定，与同窝阴性大鼠（TauT-/-大鼠）比较，TauT+/-大鼠大脑组织Slc6a6 mRNA基因表达水平显著升高，大脑、心、肝组织TauT蛋白表达水平没有显著性差异，HE染色和免疫组化结果显示TauT+/-大鼠脑组织海马区和皮层区未见异常，TauT蛋白表达没有显著性差异。结论：成功构建TauT转基因大鼠品系，为后续研究TauT在大脑中的作用提供一种较好的动物模型。

Abstract:

Objective: To establish brain-specific expression taurine transporter transgenic rats(TauT+/-rats), which can be used for the study of the effects of taurine transporter(TauT) on neurological diseases. Methods：The transgene expression vector was constructed by inserting the TauT cDNA into the downstream of PDGF promoter specifically expressed in brain tissue.The transgenic rats was established by micro injection and detected the genotype by PCR．The weight curves of 2-8 months TauT+/-ratswere measured, the weight of each organ, the visceral coefficient and the visceral brain coefficient were measured. The expression of Slc6a6 mRNA in the brain tissue of TauT+/- rats was detected by RT-PCR. Western Blotting was used to detect TauT protein expression in brain, heart and liver tissues.Immunohistochemical staining and HE staining were performed on TauT+/- rat brain cortex and hippocampus, and IPP processing Immunohistochemical images. Results：TauT cDNA transgenic vector was successfully constructed and three TauT transgenic rats were established. The expression level of Slc6a6 mRNA in brain tissue of TauT+/- rat was significantly increased, but the expression levels of TauT protein in brain, heart and liver tissues were not. HE staining and immunohistochemistry showed no abnormalities and significant difference in brain cortex and hippocampus. Conclusion：The brain-specific expression TauT transgenic rat was successfully established and identified. The transgenic rat could be a useful model to research the function of taurine transporter in brain.

参考文献/References:

[1] Pasantes-Morales H. Taurine Homeostasis and Volume Control[J].Adv Neurobiol, 2017,16:33
[2] Murakami S. Role of Taurine in the Pathogenesis of Obesity[J]. Mol Nutr Food Res, 2015,59(7):1353
[3] 刘妍妍, 崔志清. 牛磺酸对小鼠肠推进及大鼠离体回肠平滑肌运动的影响[J]. 天津医科大学学报, 2008,14(4):441
[4] Pasantes-Morales H, Hernández-Benítez R. Taurine and Brain Development: Trophic or Cytoprotective Actions[J]. Neurochem Res,2010,35(12):1939
[5] Gether U, Andersen P H, Larsson O M, et al. Neurotransmitter Transporters: Molecular Function of Important Drug Targets[J].Trends Pharmacol Sci, 2006, 27(7):375
[6] Olson J E, Martinho E Jr. Regulation of Taurine Transport in Rat Hippocampal Neurons by Hypo-Osmotic Swelling[J]. J Neurochem, 2006(96):1375
[7] Hernández-Benítez R, Pasantes-Morales H, Pinzón-Estrada E, et al. Functional expression and subcellular localization of the taurine transporter TauT in murine neural precursors[J]. Dev Neurosci, 2010,32(4):321
[8] 张小龙, 邓欢, 陈澄，等. 牛磺酸转运体的调节机制及其在动物营养研究中的意义[J]. 动物营养学报, 2013,25(10):2222
[9] Zheng S, Geghman K, Shenoy S, et al. Retake the Center Stage--New Development of Rat Genetics[J]. J Genet Genomics, 2016,39(6):261
[10] Flood D G, Lin Y G, Lang D M, et al. A Transgenic Rat Model of Alzheimer's Disease with Extracellular Abeta Deposition[J]. Neurobiol Aging, 2009, 30(7):1078
[11] Zharikov A, Bai Q, De Miranda B R, et al. Long-term RNAi knockdown of α-synuclein in the adult rat substantia nigra without neurodegeneration[J]. Neurobiol Dis, 2019,125:146
[12] Shishkina G T, Lanshakov D A, Bannova A V, et al. Doxycycline Used for Control of Transgene Expression has its Own Effects on Behaviors and Bcl-xL in the Rat Hippocampus[J]. Cell Mol Neurobiol, 2018,38(1):281
[13] 肖红卫，华文君，张立苹，等. 转基因动物技术及其在动物育种中的应用[J]. 中国猪业,2016,11(4):56
[14] 冯颖, 杨建成, 孙长勉,等.牛磺酸对不同月龄雄性大鼠生殖器官指数及生殖激素水平的影响[J]. 黑龙江畜牧兽医,2006(6):5
[15] 戴丽军, 叶炳飞, 黄月玲. Fmr1基因敲除小鼠脏器重量和脏器系数的比较分析[J]. 2009,19(6):59
[16] Branco R C, Batista T M, Camargo R L, et al. Long-Term Taurine Supplementation Leads to Enhanced Hepatic Steatosis, Renal Dysfunction and Hyperglycemia in Mice Fed on a High-Fat[J]. Adv Exp Med Biol, 2015,803:339
[17] 张丽, 孙彩显, 靳亚西，等. Dkk3过表达改善阿尔茨海默症小鼠的老年斑、认知障碍和脑部糖代谢[J]. 神经药理学报，2017,7(2):40
[18] Sasahara M, Fries J W, Raines E W， et al. PDGF B-chain in neurons of the central nervous system, posterior pituitary, and in a transgenic model[J]. Cell,1991,64(1): 217
[19] Liu B H, Wang X, Ma Y X, et al. CMV enhancer/human PDGF-beta promoter for neuron-specific transgene expression[J]. Gene Ther,2004,11(1):52
[20] Fang F, Yang W, Florio J B, et al. Synuclein impairs trafficking and signaling of BDNF in a mouse model of Parkinson’s disease[J]. Sci Rep, 2017, 7(1):3868
[21] Schreglmann S R, Regensburger M, Rockenstein E, et al. The temporal expression pattern of alpha-synuclein modulates olfactory neurogenesis in transgenic mice[J]. PLoS One, 2015,10(5):e0126261

相似文献/References:

[1]刘文东,李英娴,娄婷婷,等.青蒿琥酯对肝纤维化大鼠肝组织基质金属蛋白酶表达的影响[J].天津医科大学学报,2013,19(05):362.
[2]张文,宋红丽,杨洋,等.大鼠异位小肠移植模型手术技术的优化[J].天津医科大学学报,2013,19(05):377.
[3]刘 琪,王大为,陈 兴,等.氢气对吸入性损伤大鼠急性肺损伤的保护作用及量效研究[J].天津医科大学学报,2013,19(06):445.
　LIU Qi,WANG Da-wei,CHEN Xing,et al.Protective effects of hydrogen gas on acute lung injury induced by smoke inhalation in rats and its dose-effect relationship[J].Journal of Tianjin Medical University,2013,19(01):445.
[4]张立亚,崔尧丽,王 兵,等.z-VAD-FMK与Necrostatin-1联合对MODS大鼠器官的保护作用研究[J].天津医科大学学报,2014,20(02):85.
　ZHANG Li-ya,CUI Yao-li,WANG Bing,et al. Protective effect of z-VAD-FMK and Necrostatin-1 on the organs of rats with MODS[J].Journal of Tianjin Medical University,2014,20(01):85.
[5]于金宝,王勇强,崔尧丽,等.zVAD-fmk对大鼠肾缺血再灌注损伤的保护作用[J].天津医科大学学报,2014,20(02):89.
　YU Jin-bao,WANG Yong-qiang,CUI Yao-li,et al. Protective effects of z-VAD-FMK on renal ischemia-reperfusion injury in rat[J].Journal of Tianjin Medical University,2014,20(01):89.
[6]甘文学,张 瑞,张欣,等.大鼠小肠分泌胰岛素基因水平研究[J].天津医科大学学报,2014,20(02):98.
　GAN Wen-xue,ZHANG Rui,ZHANG Xin,et al.Research on gene level of small intestine secreting insulin in rats[J].Journal of Tianjin Medical University,2014,20(01):98.
[7]王菲,常光明,耿鑫.BDNF和TERT联合转染BMSCs对血管性痴呆大鼠学习记忆功能恢复及海马CA1区超微结构的影响[J].天津医科大学学报,2014,20(03):172.
　WANG Fei,CHANG Guang-ming,GENG Xin.BMSCs co-transfected with BDNF and TERT improve the ability of cognition and ultra-structure in the hippocampal CA1 region of the rats with vascular dementia[J].Journal of Tianjin Medical University,2014,20(01):172.
[8]徐 倩,杜洪印,喻文立,等.雷帕霉素对大鼠肝脏冷缺血再灌注损伤的保护作用[J].天津医科大学学报,2014,20(03):181.
　XU qian,DU Hong-yin,YU Wen-li,et al.Effects of rapamycin on liver cold ischemia/reperfusion injury in rats[J].Journal of Tianjin Medical University,2014,20(01):181.
[9]张 俊,孟令贺,单士军,等. 康复新液对实验性大鼠皮肤切割伤痂下愈合的影响[J].天津医科大学学报,2014,20(03):192.
　ZHANG Jun,MENG Ling-he,SHAN Shi-jun,et al.Effect of Kangfuxin liquid on subcrustal wound healing of rat[J].Journal of Tianjin Medical University,2014,20(01):192.
[10]巫 强,朱志军,徐 倩,等.含饱和氢气灌注液对大鼠肝脏冷缺血再灌注肺损伤的影响[J].天津医科大学学报,2014,20(04):267.
　WU Qiang,ZHU Zhi-jun,XU Qian,et al.Effects of hydrogen–rich perfusion on lung injury by hepatic cold ischemia／reperfusion in rat[J].Journal of Tianjin Medical University,2014,20(01):267.

备注/Memo

备注/Memo:

基金项目 国家自然科学基金面上项目（81571216）；天津市应用基础及前沿技术重点项目（16JCZDJC35500）
作者简介 齐浩铭（1994-），男，硕士在读，研究方向：生物化学与分子生物学;通信作者：黄慧玲，E-mail:huanghuiling@126.com。

常用功能

更新日期/Last Update: 2020-04-16