脑内过表达胰岛素样生长因子结合蛋白-4转基因小鼠的建立

doi:10.3969/j.issn.1006-7795.2015.04.018

首都医科大学学报 ›› 2015, Vol. 36 ›› Issue (2): 262-269.doi: 10.3969/j.issn.1006-7795.2015.04.018

脑内过表达胰岛素样生长因子结合蛋白-4转基因小鼠的建立

勾荣彬, 江小华, 巨荣凯, 牛含虚, 段德义, 徐群渊

首都医科大学基础医学院神经生物学系北京脑重大疾病研究院北京市神经再生修复研究重点实验室, 北京 100069

收稿日期:2015-01-28 发布日期:2015-04-16
通讯作者: 徐群渊 E-mail:xuqy@ccmu.edu.cn
基金资助:
国家自然科学基金(81070977,81200931,81271388),首都医科大学神经再生修复研究北京市重点实验室开放课题(2011SJZS05,2012SJZS03)。

Establishment of transgenic mice over-expressing insulin-like growth factor binding protein-4 in the brain

Gou Rongbin, Jiang Xiaohua, Ju Rongkai, Niu Hanxu, Duan Deyi, Xu Qunyuan

Department of Neurobiology, School of Basic Medical Sciences, Capital Medical University, Beijing Center of Neural Regeneration & Repair, Beijing Institute for Brain Disorders, Beijing 100069, China

Received:2015-01-28 Published:2015-04-16
Supported by:
This study was supported by National Natural Science Foundation of China(81070977, 81200931, 81271388), Capital Medical University Key Laboratory Research Fund(2011SJZS05, 2012SJZS03).

摘要/Abstract

摘要： 目的构建脑内过表达胰岛素样生长因子结合蛋白-4(insulin-like growth factor binding protein-4,IGFBP-4)的转基因小鼠,以研究IGFBP-4对脑发育的作用。方法构建神经元特异性启动子血小板源性生长因子β链(platelet derived growth factor β chain,PDGF-β)驱动的IGFBP-4表达载体,通过显微注射构建转基因小鼠。用PCR法检测小鼠子代基因组IGFBP-4 cDNA的整合情况,Western blotting法和免疫组织化学法观察IGFBP-4在脑内的表达丰度和分布,比较野生型和转基因小鼠的体质量和脑质量的改变。结果外源IGFBP-4 cDNA稳定整合于转基因小鼠基因组,IGFBP-4蛋白在成年小鼠脑内表达水平升高了267%,脑区IGFBP-4蛋白分布符合PDGF-β启动子驱动的蛋白表达特征,与野生型小鼠IGFBP-4部位相似。转基因小鼠体质量未见明显改变,整脑质量增加了8.4%(P<0.05),其中脑干增加了16.5%(P<0.05),其余脑区改变不明显。结论外源IGFBP-4转基因在小鼠基因组中得到整合并能稳定遗传和表达,并影响了部分脑区的发育,为脑发育研究提供了有价值的动物模型。

关键词: 胰岛素样生长因子结合蛋白-4, 转基因小鼠, 神经发育

Abstract: Objective To establish a transgenic mouse which may over express insulin-like growth factor binding protein-4(IGFBP-4) in the brain, in order to explore the effect of IGFBP-4 on brain development. Methods The neuron-specific transgene expression cassette of IGFBP-4 was constructed and then microinjected into the fertilized mouse egg. The genotype of the transgenic mice was identified using PCR. The expression of IGFBP-4 protein in the brain was detected by Western blotting, and the spatial expression of IGFBP-4 was detected by immunohistochemistry. Both weights of body and brain of mice were measured. Results Exogenous IGFBP-4 cDNA was successfully integrated into the mouse genome and stably passaged. The expression of IGFBP-4 protein in the adult brain was increased by 267% in the transgenic mouse, while its spatial expression pattern was similar to that in the wild type mouse. The body weight was insignificantly changeable, but the whole brain weight was found an 8.4% increase in transgenic mice(P<0.05). Especially, the weight of brain stem increased by 16.5%(P<0.05), with no significant changes in other areas of the brain. Conclusion The exogenous IGFBP-4 cDNA was stably integrated into the transgenic mouse genome, and the over-expression of IGFBP-4 could affect development of certain part of the brain. The newly established IGFBP-4 transgenic mouse may therefore provide a useful animal model for investigating brain development.

Key words: insulin-like growth factor binding protein-4(IGFBP-4), transgenic mice, neurodevelopment

中图分类号:

Q786

勾荣彬, 江小华, 巨荣凯, 牛含虚, 段德义, 徐群渊. 脑内过表达胰岛素样生长因子结合蛋白-4转基因小鼠的建立[J]. 首都医科大学学报, 2015, 36(2): 262-269.

Gou Rongbin, Jiang Xiaohua, Ju Rongkai, Niu Hanxu, Duan Deyi, Xu Qunyuan. Establishment of transgenic mice over-expressing insulin-like growth factor binding protein-4 in the brain[J]. Journal of Capital Medical University, 2015, 36(2): 262-269.

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参考文献

[1] Pera E M, Acosta H, Gouignard N, et al. Active signals, gradient formation and regional specificity in neural induction[J]. Exp Cell Res, 2014,321(1):25-31.
[2] Mohan S, Baylink D J. IGF-binding proteins are multifunctional and act via IGF-dependent and -independent mechanisms[J]. J Endocrinol, 2002,175(1):19-31.
[3] O'Kusky J, Ye P. Neurodevelopmental effects of insulin-like growth factor signaling[J]. Front Neuroendocrinol, 2012,33(3):230-251.
[4] Yu S, Zhang J Z, Xu Q. Genes associated with neuronal differentiation of precursors from human brain[J]. Neuroscience, 2006,141(2):817-825.
[5] Jiang X, Zhao J, Ju L, et al. Temporal expression patterns of insulin-like growth factor binding protein-4 in the embryonic and postnatal rat brain[J]. BMC Neurosci, 2013,14:132.
[6] 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):52-60.
[7] Paxinos G, Franklin K. The mouse brain in stereotaxic coordinates[M]. Sydney:Gulf Professional Publishing, 2004.
[8] 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):217-227.
[9] Sasahara A, Kott J N, Sasahara M, et al. Platelet-derived growth factor B-chain-like immunoreactivity in the developing and adult rat brain[J]. Brain Res Dev Brain Res, 1992,68(1):41-53.
[10] Sasahara M, Sato H, Iihara K, et al. Expression of platelet-derived growth factor B-chain in the mature rat brain and pituitary gland[J]. Brain Res Mol Brain Res, 1995,32(1):63-74.
[11] Georgopoulos S, McKee A, Kan H Y, et al. Generation and characterization of two transgenic mouse lines expressing human ApoE2 in neurons and glial cells[J]. Biochemistry, 2002,41(30):9293-9301.
[12] Brar A K, Chernausek S D. Localization of insulin-like growth factor binding protein-4 expression in the developing and adult rat brain:analysis by in situ hybridization[J]. J Neurosci Res, 1993,35(1):103-114.
[13] Stenvers K L, Zimmermann E M, Gallagher M, et al. Expression of insulin-like growth factor binding protein-4 and -5 mRNAs in adult rat forebrain[J]. J Comp Neurol, 1994,339(1):91-105.
[14] Kang D W, Chung Y H, Lee J C, et al. Immunohistochemical study on the distribution of insulin-like growth factor-binding protein 4 in the central nervous system of SOD1(G93A) transgenic mice as an in vivo model of amyotrophic lateral sclerosis[J]. Ann Anat, 2008,190(6):502-509.
[15] Ning Y, Schuller A G, Conover C A, et al. Insulin-like growth factor(IGF) binding protein-4 is both a positive and negative regulator of IGF activity in vivo[J]. Mol Endocrinol, 2008,22(5):1213-1225.
[16] Gutiérrez-Ospina G, Calikoglu A S, Ye P, et al. In vivo effects of insulin-like growth factor-I on the development of sensory pathways:analysis of the primary somatic sensory cortex(S1) of transgenic mice[J]. Endocrinology, 1996,137(12):5484-5492.
[17] Popken G J, Hodge R D, Ye P, et al. In vivo effects of insulin-like growth factor-I(IGF-I) on prenatal and early postnatal development of the central nervous system[J]. Eur J Neurosci, 2004,19(8):2056-2068.
[18] Ye P, Popken G J, Kemper A, et al. Astrocyte-specific overexpression of insulin-like growth factor-I promotes brain overgrowth and glial fibrillary acidic protein expression[J]. J Neurosci Res, 2004,78(4):472-484.
[19] Carson M J, Behringer R R, Brinster R L, et al. Insulin-like growth factor I increases brain growth and central nervous system myelination in transgenic mice[J]. Neuron, 1993,10(4):729-740.
[20] Ye P, Carson J, D'Ercole A J. Insulin-like growth factor-I influences the initiation of myelination:studies of the anterior commissure of transgenic mice[J]. Neurosci Lett, 1995,201(3):235-238.
[21] Beck K D, Powell-Braxton L, Widmer H R, et al. Igf1 gene disruption results in reduced brain size, CNS hypomyelination, and loss of hippocampal granule and striatal parvalbumin-containing neurons[J]. Neuron, 1995,14(4):717-730.
[22] Qin X, Strong D D, Baylink D J, et al. Structure-function analysis of the human insulin-like growth factor binding protein-4[J]. J Biol Chem, 1998,273(36):23509-23516.

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脑内过表达胰岛素样生长因子结合蛋白-4转基因小鼠的建立

Establishment of transgenic mice over-expressing insulin-like growth factor binding protein-4 in the brain

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