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Construction of Eukaryotic Expression Vectors of ERBB4 and NRG1 Genes and Its Expression in Cairna moschata Nerve Cells |
LUO Lin-Li1, YANG Sheng-Lin1,*, ZHOU Xuan1, ZHANG Yun2, YANG Shi-Hao1, ZHU Yong-Cai1 |
1 College of Animal Science, Guizhou University/Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region, Ministry of Education, Guiyang 550025, China; 2 Guizhou Province Livestock and Poultry Genetic Resources Management Station, Guiyang 550025, China |
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Abstract Receptor tyrosine protein kinase 4 (ERBB4) and neuregulin 1 (NRG1) genes play an important role in the nervous system, the expression of ERBB4 gene in aggressive Cairna moschata was higher than that in normal group. In order to further explore the ERBB4 and NRG1 genes expression in nerve cells of Cairna moschata, pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 eukaryotic expression vectors were built and transfected to nerve cells of Cairna moschata, then the expression levels of ERBB4 and NRG1 genes and the downstream Src homolog and collagen homolog (Shc) protein and growth factor receptor bound protein 2 (Grb2) in the NRG1-ERBB4 signaling pathway were detected by qRT-PCR. The results showed that the expression of ERBB4 and NRG1 in the transfected pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 cells was significantly higher than those of pEGFP-C1 and pDsRED-C1 empty vector cells (P<0.01), indicating that pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 were successfully overexpressed in nerve cells of Cairna moschata; The expression of NRG1 in transfected pEGFP-C1-ERBB4 cells was significantly higher than that in transfected pEGFP-C1 empty vector cells (P<0.01), and the expression of ERBB4 in transfected pDsRED-C1-NRG1 cells was significantly higher than that in transfected pEGFP-C1 empty vector cells (P<0.01), it showed that the overexpression of ERBB4 and NRG1 both the were up-regulated by each other; Compared with the transfected pEGFP-C1 and pDsRED-C1 empty vector and the blank experimental group, the expression of Shc and Grb2 in transfected pEGFP-C1-ERBB4 and pDsRED-C1-NRG1 cells were significantly up-regulated (P<0.01). The above results indicated that the expression up-regulated of ERBB4 or NRG1 significantly increased the expression of the other one, and the downstream genes Shc and Grb2 of NRG1-ERBB4 signaling pathway were increased in Cairna moschata nerve cells. The results of this study provide a basis for further exploring the mechanism of ERBB4 and NRG1 genes in the aggressive behavior of Cairna moschata.
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Received: 02 July 2020
Published: 01 May 2021
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Corresponding Authors:
*shenglinyang@126.com
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[1] 陈杰, 陈策, 刘立滢, 等. 2019. α-葎草烯调控NRG1/ErbB4信号通路改善精神分裂症研究[J]. 中草药, 50(23): 5790-5794. (Chen J, Chen C, Liu L Y, et al.2019. Improvement of schizophrenia by regulating the NRG1/ErbB4 signaling pathway[J]. Chinese Herbal Medicine, 50(23): 5790-5794.) [2] 樊新龙, 郭囡, 高鑫, 等. 2019. 上调NRG1对甲状腺癌细胞增殖和凋亡的影响及可能机制[J]. 现代肿瘤医学, 27(01): 21-25. (Fan X L, Guo N, Gao X, et al.2019. Effects of up-regulation of NRG1 on proliferation and apoptosis of thyroid cancer cells and its possible mechanism[J]. Modern Oncology Medicine, 27(01): 21-25.) [3] 郭汝金, 敖丽娟, 李咏梅, 等. 2014. 神经调节蛋白1-ErbB信号通路的研究进展[J]. 中国康复医学杂志, 29(07): 679-684. (Guo R J, Ao L J, Li Y M, et al.2014. The neural regulation protein 1-the research progress of ErbB signaling pathways[J]. Chinese Journal of Rehabilitation Medicine, 29(7): 679-684.) [4] 何海洋. 2017. NRG1/ErbB4信号通路在MeCP2缺陷小鼠异常行为中作用的研究[D]. 硕士学位论文, 浙江大学, 导师: 李晓明, pp. 18-19. (He H Y.2017. The role of NRG1/ErbB4 signaling pathway in abnormal behavior of MeCP2 deficient mice[D]. Thesis for M.S., Zhejiang University, supervisor: Li X M , pp. 18-19.) [5] 李明如, 聂振凯, 林贵湖, 等. 2019. ERBB4通过靶向调控miR-193a-3p调节TP53INP1的表达[J].生物技术通讯, 30(05): 624-629. (Li M R, Nie Z K, Lin G H, et al.2019. ERBB4 regulates TP53INP1 expression by targeting Mir-193A-3P[J]. Biotechnology Communications, 30(05): 624-629.) [6] 李树玲, 颜慧, 宫泽辉. 2013. ErbB4基因miRNA慢病毒载体的构建及其在小鼠海马齿状回的表达[J]. 中国药理学与毒理学杂志, 27(01): 29-33. (Li S L, Yan H, Gong Z H.2013. Construction of ErbB4 gene miRNA lentiviral vector and its expression in mouse hippocampal dentate gyrus[J]. Chinese Journal of Pharmacology and Toxicology, 27(01): 29-33.) [7] 吉文林, 段修军, 秦豪荣, 等. 2008. 黑羽番鸭与法国番鸭生长及生产性能的比较研究[J]. 黑龙江畜牧兽医, (04): 48-50. (Ji W L, Duan X J, Qin H R, et al. 2008. Comparative study on growth and production performance of Black feather Muscovette and French muscovette[J]. Heilongjiang Animal Husbandry and Veterinary Science, (04): 48-50.) [8] 孙娇丽, 王鹏, 伍宗芳, 等. 2014. NRG1-siRNA慢病毒载体的构建[J]. 中华麻醉学杂志, 34(02): 164-166. (Sun J l, Wang P, Wu Z F, et al.2014. Construction of nrg1-sirna lentiviral vector[J]. Chinese Journal of Anesthesiology, 34(02): 164-166.) [9] 王凯, 闫磊, 刘志新, 等. 2017. NRG1转染对骨髓基质干细胞表达VEGF的影响[J]. 中国当代医药, 24(32): 4-7. (Wang K, Yan L, Liu Z X, et al.2017. Effect of NRG1 transfection on VEGF expression in bone marrow stromal stem cells[J]. Chinese Journal of Contemporary Medicine, 24(32): 4-7.) [10] 王楠. 2014. NRG1-ErbB4信号通路在氯胺酮抗抑郁中的作用[D]. 硕士学位论文, 南京大学, 导师: 杨建军, pp. 14-16. (Wang N.2014. The role of NRG1-ERBB4 signaling pathway in ketamine anti-depression[D]. Thesis for M.S., Nanjing University, Supervisor: Yang J J, pp. 14-16.) [11] 王旭平. 2018. 番鸭攻击行为关联基因的转录组学分析[D]. 硕士学位论文, 贵州大学, 导师: 杨胜林, pp. 49-50. (Wang X P.2018. Transcriptional analysis of genes associated with aggression in Muscovus chinensis[D]. Thesis for M.S., Guizhou University, Supervisor: Yang S L, pp. 49-50.) [12] 闫竞一. 2017. 伊马替尼继发性耐药胃肠间质瘤中ErbB信号通路的变化[D]. 博士学位论文, 苏州大学, 导师: 钱海鑫, pp. 33-35. (Yan J Y.2017. Changes of ErbB signaling pathway in imatinib resistant gastrointestinal stromal tumors[D]. Thesis for Ph.D., Suzhou University, supervisor: Qian H X, pp. 33-35.) [13] 周璇, 杨胜林, 杨世皓, 等. 2020. 番鸭ERBB4基因组织表达、克隆及序列分析[J/OL]. 中国畜牧兽医, 47(06): 1637-1647. (Zhou X, Yang S L, Yang S H, et al.2020. Expression, cloning and sequence analysis of ERBB4 gene in Muscovag[J/OL]. Chinese Animal Husbandry and Veterinary Science, 47(06): 1637-1647.) [14] Ananloo E S, Yoosefee S, Karimipour M.2020. Neuregulin1 gene variants as a biomarker for cognitive impairments in patients with schizophrenia[J]. The European Journal of Psychiatry, 34(1): 11-19. [15] Almeida R M, Ferrari P F, Parmigiani S, et al.2005. Escalated aggres-sive behavior: Dopamine, serotonin and GABA[J]. European Journal of Pharmacology, 526(1-3): 51-64. [16] Bannai M, Fish E W, Faccidomo S, et al.2007. Anti-aggressive effects of agonists at 5-HT1B receptors in the dorsal raphe nucleus of mice[J]. Psychopharmacology (Berl), 193(2): 295-304. [17] Bilsing A, Holub H, Nichelmann M.1993. Sexual behavior of muscovy ducks under conditions of intensive husbandry of adult animals: Dependence on the time of day, husbandry conditions, body mass and health status[J]. Berliner und Münchener tierärztliche Wochenschrift, 106(2): 55-59. [18] Chung D W, Chung Y, Bazmi H H, et al.2018. Altered ErbB4 splicing and cortical parvalbumin interneuron dysfunction in schizophrenia and mood disorders[J]. Neuropsychopharmacology, 43(8): 2478-2486. [19] Deng W k, Luo F, Li B M, et al.2019. NRG1-ErbB4 signaling promotes functional recovery in a murine model of traumatic brain injury via regulation of GABA release[J]. Experimental Brain Research, 237(12): 3351-3362. [20] Dixon M, Lumsden A.1999. Distribution of neuregulin-1 (nrg1) and erbB4 transcripts in embryonic chick hindbrain[J]. Molecular and Cellular Neuroence, 13(4): 237-258. [21] Huber D, Veinante P, Stoop R.2005. Vasopressin and oxytocin excite distinct neuronal populations in the central amygdala[J]. Science, 308(5719): 245-248 [22] Karl T, Lin S, Schwarzer C, et al.2004. Y1 receptors regulate aggressive behavior by modulating serotonin pathways[J]. Proceedings of the National Academy of Sciences of USA, 101(34): 12742-12747. [23] Mei L, Xiong W C.2008. Neuregulin 1 in neural development, synaptic plasticity and schizophrenia[J]. Nature Reviews. Neuroscience, 9(6): 437-452 [24] Shin Y K, Jang S Y, Park S Y, et al.2014. Grb2-associated binder-1 is required for neuregulin-1-induced peripheral nerve myelination[J]. Journal of Neuroscience, 34(22): 7657-7622. [25] Wang J, Song F, Loeb J A.2017. Neuregulin1 fine-tunes pre-, post-, and perisynaptic neuromuscular junction development[J]. Developmental Dynamics, 246(5): 368-380. [26] Wang F, Jiang T, Sun Z, et al.2009. Neuregulin 1 genetic variation and anterior cingulum integrity in patients with schizophrenia and healthy controls[J]. Journal of Psychiatry & Neuroscience, 34(3): 181-186. [27] Zhang C C, Ni P Y, Liu Y K, et al.2020. GABAergic abnormalities associated with sensorimotor cortico-striatal community structural deficits in ErbB4 knockout mice and first-episode treatment-naïve patients with schizophrenia[J]. Neuroscience Bulletin, 36(02): 97-109. [28] Zeng Z, Gui C, Nong Q, et al.2013. Serum neuregulin-1β levels are positively correlated with VEGF and angina pectoris[J]. International Journal of Cardiology, 168(3): 3077-3079. |
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