<em>ERBB4</em>、<em>NRG1</em>基因真核表达载体构建及其在番鸭神经细胞中的表达

doi:10.3969/j.issn.1674-7968.2021.05.012

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摘要酪氨酸激酶受体4 (receptor tyrosine protein kinase 4, ERBB4)、神经调节蛋白1 (neuregulin 1, NRG1)基因在神经系统中都发挥着重要作用,且ERBB4基因在攻击行为番鸭(Cairna moschata)脑组织中的表达量较高。为进一步探索ERBB4、NRG1基因在番鸭神经细胞中的表达情况,构建pEGFP-C1-ERBB4和pDsRED-C1-NRG1真核表达载体,将其转染番鸭神经细胞,运用qRT-PCR技术检测ERBB4、NRG1基因表达量和NRG1-ERBB4信号通路的下游基因衔接蛋白基因(Src homolog and collagen homolog, Shc)、生长因子受体结合蛋白2基因(growth factor receptor bound protein 2, Grb2)的表达情况。结果显示,转染pEGFP-C1-ERBB4、pDsRED-C1-NRG1细胞中ERBB4、NRG1表达极显著高于转染pEGFP-C1、pDsRED-C1空载体细胞(P<0.01),表明pEGFP-C1-ERBB4、pDsRED-C1-NRG1成功过表达于鸭神经细胞;转染pEGFP-C1-ERBB4细胞中NRG1表达极显著高于转染pEGFP-C1空载体细胞(P<0.01),转染pDsRED-C1-NRG1细胞中ERBB4表达极显著高于转染pDsRED-C1空载体细胞(P<0.01),表明ERBB4、NRG1过量表达均相互促进彼此上调表达;与转染pEGFP-C1和pDsRED-C1空载体和空白实验组相比,转染pEGFP-C1-ERBB4、pDsRED-C1-NRG1细胞中Shc、Grb2表达均极显著上调(P<0.01)。本研究结果表明在番鸭神经细胞中ERBB4或NRG1表达上调均显著增加对方的表达,并提高NRG1-ERBB4信号通路下游基因Shc、Grb2的表达。本研究结果为进一步探讨ERBB4、NRG1基因在番鸭攻击行为中的作用机制提供了一定基础依据。

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	罗林丽
	杨胜林
	周璇
	张芸
	杨世皓
	祝永才

关键词 ：番鸭, ERBB4基因, NRG1基因, 表达, 攻击行为

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.

Key words： Muscovy duck ERBB4 gene NRG1 gene Expression Aggressive behavior

收稿日期: 2020-07-02 出版日期: 2021-05-01

ZTFLH:

S813

基金资助:国家自然科学基金(31960682); 贵州省科技厅平台项目(黔科合平台人才[2019]5203]); 中央引导地方科技发展专项资金项目(黔科中引地[2019]4021); 贵州省第六批人才基地项目((黔人领发)[2018]3号); 贵州省普通本科高等学校科技成果转化与产业化项目(黔教合KY字[2017]056); 贵州省科技成果转化项目(黔科合[2020]1Y041)

通讯作者: *shenglinyang@126.com

引用本文:

罗林丽, 杨胜林, 周璇, 张芸, 杨世皓, 祝永才. ERBB4、NRG1基因真核表达载体构建及其在番鸭神经细胞中的表达[J]. 农业生物技术学报, 2021, 29(5): 952-962.
LUO Lin-Li, YANG Sheng-Lin, ZHOU Xuan, ZHANG Yun, YANG Shi-Hao, ZHU Yong-Cai. Construction of Eukaryotic Expression Vectors of ERBB4 and NRG1 Genes and Its Expression in Cairna moschata Nerve Cells. 农业生物技术学报, 2021, 29(5): 952-962.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.05.012 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I5/952

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